针刺太冲与非经非穴对自发性高血压大鼠血压及延髓蛋白表达的影响
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摘要
目的
探讨针刺太冲与非经非穴对自发性高血压大鼠(SHR)血压的影响,并利用二维凝胶电泳和质谱技术分离并鉴定各组大鼠延髓内表达差异的蛋白,以期从蛋白质整体水平探讨针刺太冲穴降低SHR高血压的延髓内分子机制,并为寻找与经穴效应特异性相关的生物效应指标提供线索。方法
选用9w、雄性SHR30只,随机分为模型组、太冲组、和非经非穴组(均n=10),并以同体重正常血压SD大鼠(n=10)作为正常组。太冲组为针刺大鼠双侧太冲穴(定位在大鼠后肢足背侧第1、2跖骨间),非经非穴组为针刺大鼠双侧非经非穴(定位在大鼠后肢足背侧第3、4跖骨间凹陷处)。针刺参数为大鼠清醒状态下,针刺1次／d,持续时间5min／次,捻转频率80±5次／min,捻转幅度180±5度,太冲组毫针斜向涌泉,不穿透、不触及足底皮肤,而非经非穴组毫针垂直于皮肤进针并穿透足底皮肤,于针刺结束5min后采用BP-6无创血压测量系统对大鼠进行尾动脉血压测定。模型组和正常组不给予针刺。连续针刺治疗到第7d后,对各组大鼠断头取脑,在冰上分离获取延髓。然后裂解、超声破碎延髓组织,经除盐处理,通过二维凝胶电泳分离蛋白,Typhoon9200扫描仪获得凝胶图像,Imagemaster 2D 6.0图像分析软件分析各组蛋白差异点,用MALDI-TOF-TOF串联质谱仪分析蛋白质,所得的质谱图通过GPS软件在MASCOT数据库中检索并鉴定蛋白质,再利用酶联免疫分析实验进一步验证差异蛋白的变化。结果
1.针刺太冲与非经非穴对SHR血压的影响。组间比较结果：针刺前收缩压即原始收缩压的比较,太冲组、非经非穴组及模型组之间大鼠收缩压均无显著性差异(均P>0.05)。于针刺第1d,太冲组、非经非穴组及模型组之间大鼠收缩压均无显著性差异(均P>0.05)。于针刺第2d,与非经非穴组、模型组比较,太冲组大鼠收缩压显著降低(P<0.01),提示于针刺第2 d,太冲组大鼠收缩压开始降低。于针刺第3 d,与模型组比较,非经非穴组大鼠收缩压显著升高,提示针刺非经非穴可能是一种非良性刺激,导致大鼠收缩压显著升高。于针刺第4d,太冲组、非经非穴组及模型组之间大鼠收缩压均无显著性差异(均P>0.05)。于针刺第5d,与非经非穴组比较,太冲组大鼠收缩压显著降低(P<0.01)。于针刺第6、第7d,与非经非穴组、模型组比较,太冲组大鼠收缩压显著降低(均P<0.01),提示针刺太冲穴于第5d开始稳定的降压水平。与正常组比较,不同时间段太冲组、非经非穴组以及模型组大鼠收缩压均显著性升高(均P<0.01),提示本实验针刺太冲穴和非经非穴都未能使收缩压恢复至正常水平。
组内比较结果：与第1d比较,太冲组大鼠收缩压于第2d开始显著降低(P<0.01),然而于第3、第4d收缩压有小幅度升高(P>0.05),然后又于第5d开始降低(P<0.01),直到第6d、第7d收缩压降低水平保持稳定(均P<0.01),提示针刺太冲穴于第5d开始稳定的降压水平。非经非穴组于不同时间段收缩压均无显著性差异(P>0.05)。与第3d相比,模型组大鼠收缩压于第6d显著升高(P<0.01),与第4d比较,模型组大鼠收缩压于第6d和第7d均显著升高(均P<0.01),提示模型组大鼠的收缩压呈上升趋势。正常组于不同时间段收缩压均无显著性差异(P>0.05)。
2.各组大鼠延髓二维凝胶电泳图谱及其分析结果。本实验建立了正常SD大鼠、SHR以及对SHR针刺太冲与非经非穴后延髓的二维凝胶电泳图谱,共发现了SD大鼠延髓有621±30个蛋白点,SHR延髓有621±20个蛋白点,SHR经针刺太冲后延髓有621±40个蛋白点,SHR经针刺非经非穴后延髓有621±20个蛋白点,其中,与SHR组比较,SHR经针刺太冲后延髓表达有差异的蛋白点88个(大于2倍)；SHR经针刺非经非穴后延髓表达有差异的蛋白点91个(大于2倍),正常SD大鼠延髓表达有差异的蛋白点47个(大于2倍)。
3.鉴定出的差异蛋白质在各组大鼠延髓表达的结果。(1)与模型组比较,太冲组大鼠延髓下调的蛋白有5个(大于2倍)：突触素蛋白1,NAD依赖的脱乙酰酶,蛋白激酶C抑制蛋白,电子转移黄素蛋白和髓磷脂碱蛋白；上调的蛋白有6个(大于2倍)：谷氨酸脱氢酶,谷胱甘肽S转移酶,醛脱氢酶2, Rho GDP解离抑制因子,DJ-1蛋白和过氧化物歧化酶。(2)与模型组比较,非经非穴组大鼠延髓下调的蛋白有1个(大于2倍)：电子转移黄素蛋白；上调的蛋白有3个(大于2倍)：热休克蛋白90,泛素水解酶和DJ-1蛋白。(3)与非经非穴组比较,太冲组大鼠延髓下调的蛋白有7个(大于2倍)：热休克蛋白90,突触素蛋白1,丙酮酸激酶,NAD依赖的脱乙酰酶,蛋白激酶C抑制蛋白1,泛素水解酶和髓磷脂碱蛋白；上调的蛋白有6个(大于2倍)：谷氨酸脱氢酶,醛脱氢酶2,谷胱甘肽S转移酶,Rho GDP解离抑制因子,DJ-1蛋白及过氧化物歧化酶。(4)与正常组比较,模型组大鼠延髓下调的蛋白6个(大于2倍)：丙酮酸激酶,谷氨酸脱氢酶,谷胱甘肽S转移酶,醛脱氢酶2,泛素水解酶和过氧化物歧化酶；上调的蛋白有7个(大于2倍)：热休克蛋白90,突触素蛋白1,NAD依赖的脱乙酰酶,电子转移黄素蛋白,蛋白激酶C抑制蛋白1,蛋白激酶C抑制蛋白1和髓磷脂碱蛋白。(5)与正常组比较,太冲组大鼠延髓下调的蛋白有3个(大于2倍)：过氧化物歧化酶,过氧化物歧化酶和髓磷脂碱蛋白；上调的蛋白有4个(大于2倍)：热休克蛋白90, Rho GDP解离抑制因子,DJ-1蛋白和过氧化物歧化酶。(6)与正常组比较,非经非穴组大鼠延髓下调的有4个(大于2倍)：谷氨酸脱氢酶,醛脱氢酶2,谷胱甘肽S转移酶和过氧化物歧化酶；上调的蛋白有7个(大于2倍)：热休克蛋白90,突触素蛋白,NAD依赖的脱乙酰酶,蛋白激酶C抑制蛋白1,泛素水解酶,DJ-1蛋白和髓磷脂碱蛋白。
5.用酶联免疫分析结果。(1)各组大鼠延髓谷胱甘肽S转移酶表达的变化。模型组大鼠延髓谷胱甘肽S转移酶表达显著低于正常组(P<0.01)。经针刺治疗后,太冲组大鼠延髓谷胱甘肽S转移酶表达显著高于模型组、非经非穴组(均P<0.01),与正常组比较无统计学差异(P>0.01),而非经非穴组大鼠延髓谷胱甘肽S转移酶表达与模型组比较无统计学差异(P>0.01),且低于正常组(P<0.01)。(2)各组大鼠延髓醛脱氢酶表达的变化。模型组大鼠延髓醛脱氢酶表达显著低于正常组(P<0.01)。经针刺治疗后,太冲组大鼠延髓醛脱氢酶表达显著高于模型组、非经非穴组(均P<0.01),而非经非穴组大鼠延髓醛脱氢酶表达与模型组比较无统计学差异(P>0.01),且低于正常组(P<0.01)。(3)各组大鼠延髓蛋白激酶C表达的变化。模型组大鼠延髓蛋白激酶C表达与正常组无显著性差异(P>0.01)。经针刺治疗后,太冲组大鼠延髓蛋白激酶C表达无明显变化,与模型组、正常组、非经非穴组比较(均P>0.01)。谷胱甘肽S转移酶、醛脱氢酶的ELISA结果与二维凝胶电泳结果一致,蛋白激酶C的ELISA结果与二维凝胶电泳蛋白激酶C抑制蛋白1的结果不一致。
结论
1.针刺太冲穴可显著降低SHR高血压,但未能使SHR高血压恢复至正常水平。
2.针刺非经非穴降低SHR高血压效果不明显,甚至可能异常升高SHR高血压。
3.针刺太冲可下调SHR延髓内促进兴奋性神经递质释放、诱发炎症反应、氧化磷酸化及信号转导相关蛋白的表达,上调氨基酸氧化分子伴侣、抑制氧化应激及信号转导相关蛋白的变化,其作用机制是多途径、多环节、多靶点的综合效应。
4.针刺非经非穴可下调SHR延髓内能量代谢相关蛋白的表达,上调氨基酸氧化分子伴侣、蛋白质分解代谢以及诱发炎症反应相关蛋白的表达,其作用机制是较少途径、较少环节、较少靶点的简单效应。
5.经穴效应及其作用机制存在一定的特异性。
Objective
In order to research the effect of acupuncture at TaiChong point (LR3) and non-acupoint on the blood pressure in Spontaneously Hypertensive Rats (SHR), and investigate, in whole level of protein expression, the central nerve mechanism of acupuncturing TaiChong point (LR3) to treat SHR, which provide the experimental clue for searching the specific indicators of biological effects that connect to the meridian effect, two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technologies were used and the differentially expressed proteins in the medulla oblongata of SHR were also separated and identified.
Methods
309-week-old male SHR were randomly divided into model group, TaiChong group and non-acupoint group, each group involve 10 SHR. And another normal control group involves 10 SD rats of normal blood pressure with the same weight with SHR. In the TaiChong group, acupuncture bilateral Taichong points (located between the 1st and the 2nd metatarsal of dorsal hind); while in non-acupoint group acupuncture bilateral non-acupoint (located at the indentation between the 3rd and 4th metatarsal of dorsal hind). As rats awake, in Taichong group and non-acupoint group, daily acupuncture 5 minutes per times, twisting nedlle 80±5 times/min frequently, Twist nedlle 180±5 degrees in rang. In Taichong group, the needle towards Yongquang but not penetrate or reach the plantar skin, rather than in non-acupoint group, needle perpendicular to the skin and penetrate the skin.5 minutes after acupuncture, using BP-6 non-invasive blood pressure measuring system for tail arterial blood pressure measurement in rats. Acupuncture treatment continnues totally 7 weeks, all the rats brains were removed at the end, then separated in the ice for the medulla oblongata. And the medulla oblongata is cracked, to Sonication, desalted, seperated for proteins with the 2-DE. Geting the gel images with Typhoon9200 scanner, analyzing the differences between the various groups of protein with Imagesaster 6.0. With MALDI-TOF-TOF tandem MS analysis, protein mass image was obtained. And with the GPS software, to search and identify protein in the MASCOT database identification, then used enzyme-linked immunosorbent assay (ELISA) test to further verify the identification of the protein.
Results
1. The influence of acupuncture at Taichong point (LR3) or non-acupoints on the blood pressure SHR'blood pressure. (1) Comparison results among groups: The original, before acupuncture, systolic blood pressure of Taichong group, and the non-acupoint group and model group were not significantly different (all P> 0.05). The 1st day in acupuncture, in Taichong group, and the non-acupoint group and model group, systolic blood pressure between the rats were not significantly different (all P> 0.05). The next day in acupuncture, compared with the model group and non-acupoints group, systolic blood pressure of Taichong group significantly decreased (P<0.01), prompting that systolic blood pressure of Taichong group started to decrease in the next day in acupuncture. The 3rd day in acupuncture, compared with model group, systolic blood pressure of the non-acupoint group were significantly higher, suggesting that the acupuncture at non-acupoint may be a non-benign stimulus, resulting in a significantly increased of systolic blood pressure in rats. On the 4th in acupuncture, systolic blood pressure of Taichong group and the non-acupoint group and model group were not significantly different (all P> 0.05). On the 5th day of acupuncture, compared with non-acupoint, Taichong group's blood pressure decreased significantly (P<0.01). In acupuncture 6th, 7th day, compared with the model group and non-acupoints group, Taichong group' systolic blood pressure were significantly lower (P<0.01), suggested acupuncture at Taichong points begin to stabilize the blood pressure level in the 5th. In different time periods, in the model group, Taichong group and non-Meridian Group, systolic blood pressure were significantly higher than normal group (P<0.01), prompted acupuncture at Taichong or non-acupoint were not able to restore blood pressure to normal levels in this study.(2) Comparison results in each group: Compared with the 1st day, Taichong group systolic blood pressure decreased significantly in the next day (P<0.01); but on the 3rd and the 4th day, the systolic blood pressure were increased in small rate (P> 0.05), and then began to decrease at the 5th day (P<0.01), until the 6th day,7th day the reducing of systolic blood pressure remained stable (P<0.01), suggested acupuncture at Taichong points begin to stabilize the blood pressure level in the 5th. Non-acupoints group'systolic blood pressure were not significantly different in different time (P> 0.05). Compared with the 3rd day, the model rats'systolic blood pressure significantly increased on the 6th day (P<0.01); compared with the 4th day, model group's systolic blood pressure were significantly increased in the 6th day and the 7th days (P<0.01), prompting that systolic blood pressure of model group was rising. In different time, in nomal group, systolic blood pressure between the rats were not significantly different (all P> 0.05).
2. The two-dimensional gel electrophoresis (2-DE) image of rat's medulla in each group, and its analysis. In this study,2-DE image were successfully obtained for normal SD rats, SHR and the SHR after acupuncturing Taichong or non-acupoint; and 621±30 proteins in normal SD rats'medulla,621±20 proteins in SHR's medulla, 621±40 proteins in medulla of SHR after acupuncturing Taichong,621±20 proteins in medulla of SHR after acupuncturing non-acupoint were found also. Compared with SHR in model group,88 proteins were found differentially expressed in medulla from Taichong group (expression ratio of protein was beyond 2 folds); 91 proteins were found differentially expressed in medulla oblongata from non-acupoints group (expression ratio of protein was beyond 2 folds),47 proteins were found differentially expressed in medulla oblongata from normal group (expression ratio of protein was beyond 2 folds).
3. Differences in protein expression in the medulla of rats that dentified. (1) In medulla of Taichong group, compared with model group,5 proteins were down-regulated (expression ratio of protein was beyond 2 folds):Synapsin-1, NAD dependent deacetylase, electron transfer flavin protein and myelin basic protein; 6 proteins were up-regulated (expression ratio of protein was beyond 2 folds): glutamate dehydrogenase, glutathione S transferase, aldehyde dehydrogenase 2, Rho GDP dissociation inhibitor, DJ-1 protein and superoxide dismutase. (2) In medulla of non-acupoint group, compared with model group,1 proteins were up-regulated (expression ratio of protein was beyond 2 folds):electron transfer flavoprotein; 3 proteins were down-regulated (expression ratio of protein was beyond 2 folds): Heat shock protein 90, the Ubiquitin hydrolase isozyme and DJ-1 protein. (3)In medulla of Taichong group, compared with non-acupoints group,7 proteins were down-regulated (expression ratio of protein was beyond 2 folds):Heat shock protein 90, Synapsin-1, pyruvate kinase, NAD dependent deacetylases enzyme, protein kinase C inhibitor protein 1, Ubiquitin hydrolase isozymeand myelin basic protein; 6 proteins were up-regulated (expression ratio of protein was beyond 2 folds):glutamate dehydrogenase, aldehyde dehydrogenase 2, glutathione S transferase, Rho GDP dissociation inhibitor, DJ-1 protein and superoxide dismutase. (4) In medulla of model group, compared with normal group,6 proteins were down-regulated (expression ratio of protein was beyond 2 folds):pyruvate kinase, glutamate dehydrogenase, glutathione S transferase, aldehyde dehydrogenase 2, Ubiquitin hydrolase isozyme and superoxide dismutase; 7 proteins were up-regulated (expression ratio of protein was beyond 2 folds):Heat shock protein 90, Synapsin-11, NAD dependent deacetylase, electron transfer flavin protein, protein kinase C inhibitor protein 1, and myelin basic protein. (5) In medulla of Taichong group, compared with normal group,3 proteins were down-regulated (expression ratio of protein was beyond 2 folds):superoxide dismutase and myelin basic protein;
4 proteins were up-regulated (expression ratio of protein was beyond 2 folds): Heat shock protein 90, Rho GDP dissociation inhibitor, DJ-1 protein and superoxide dismutase. (6) In medulla of non-acupoint group, compared with the normal group, 4 proteins were down-regulated (expression ratio of protein was beyond 2 folds): glutamate dehydrogenase, aldehyde dehydrogenase 2, glutathione S transferase and over superoxide dismutase; 7 proteins were up-regulated (expression ratio of protein was beyond 2 folds):Heat shock protein 90, synapsin 1, NAD dependent deacetylase, protein kinase C inhibitor protein 1, Ubiquitin hydrolase isozyme, DJ-1 protein and myelin basic protein.
5. results of ELISA
(1) The expression changes of glutathione S transferase in medullary among each group. In the medulla model group, compared with normal group, the expression of glutathione S transferase was significantly down-regulated (P<0.01). After acupuncture treatment, the expression of glutathione S transferase in the medulla of Taichong group were significantly up-regulated, compared with the model group, and the non-acupoint group, (P<0.01); but the expression of glutathione S transferase in the medulla of non-acupoint group has no significant difference form the model group (P> 0.01), and lower than the normal group (P<0.01). (2) The expression changes of aldehyde dehydrogenase in the medulla among each group. In the medulla model group, compared with normal group, the expression of aldehyde dehydrogenase was significantly down-regulated (P<0.01). After acupuncture treatment, the expression of aldehyde dehydrogenase in the medulla of Taichong group were significantly up-regulated, compared with the model group, and the non-acupoint group, (P<0.01); but the expression of aldehyde dehydrogenase in the medulla of non-acupoint group has no significant difference form the model group (P> 0.01), and lower than the normal group (P<0.01) (3) The expression changes of protein kinase C in medullary among each group. The expression of protein kinase C in the medulla of model group has no significant difference form the nomal group (P> 0.01); After acupuncture treatment, expression of protein kinase C in medullary of Taichong group has no significant changes from the model group, normal group, and the non-acupoint group (all P> 0.01). ELISA results of Glutathione S transferase, aldehyde dehydrogenase are consistent with the two-dimensional gel electrophoresis; meanwhile the ELISA results of protein kinase C are inconsistent with the results of two-dimensional gel electrophoresis of protein kinase C inhibitor protein 1.
Conclusion
1. Acupuncture Taichong can reduce the hypertension of SHR, but were not able to restore blood pressure to normal levels in this study.
2. acupuncture at non-acupoints didn't have an obvious effect on decreasing the blood pressure in SHR, even cause increase in SHR hypertension.
3. Acupuncture Taichong Point can down-regulated expressions of the proteins that promote excitatory neurotransmitter, induced inflammatory response, oxidative phosphorylation and induce oxidative stress protein, and up-regulate amino acid oxidation chaperones and energy metabolism and oxidative stress in SHR' medulla. Its mechanism is a multi-channel, multi-link, the combined effect of multiple targets.
4. Acupuncture non-acupoint can down-regulated expressions of the proteins that promote energy metabolism, and up-regulate amino acid oxidation, protein catabolism and expressions of protein that induced inflammatory response in medullary of SHR. The mechanism is less means, fewer links, fewer target Point of simple effect.
5. Acupoints effect and its mechanism has some specificity.
探讨针刺太冲与非经非穴对自发性高血压大鼠(SHR)血压的影响,并利用二维凝胶电泳和质谱技术分离并鉴定各组大鼠延髓内表达差异的蛋白,以期从蛋白质整体水平探讨针刺太冲穴降低SHR高血压的延髓内分子机制,并为寻找与经穴效应特异性相关的生物效应指标提供线索。方法
选用9w、雄性SHR30只,随机分为模型组、太冲组、和非经非穴组(均n=10),并以同体重正常血压SD大鼠(n=10)作为正常组。太冲组为针刺大鼠双侧太冲穴(定位在大鼠后肢足背侧第1、2跖骨间),非经非穴组为针刺大鼠双侧非经非穴(定位在大鼠后肢足背侧第3、4跖骨间凹陷处)。针刺参数为大鼠清醒状态下,针刺1次／d,持续时间5min／次,捻转频率80±5次／min,捻转幅度180±5度,太冲组毫针斜向涌泉,不穿透、不触及足底皮肤,而非经非穴组毫针垂直于皮肤进针并穿透足底皮肤,于针刺结束5min后采用BP-6无创血压测量系统对大鼠进行尾动脉血压测定。模型组和正常组不给予针刺。连续针刺治疗到第7d后,对各组大鼠断头取脑,在冰上分离获取延髓。然后裂解、超声破碎延髓组织,经除盐处理,通过二维凝胶电泳分离蛋白,Typhoon9200扫描仪获得凝胶图像,Imagemaster 2D 6.0图像分析软件分析各组蛋白差异点,用MALDI-TOF-TOF串联质谱仪分析蛋白质,所得的质谱图通过GPS软件在MASCOT数据库中检索并鉴定蛋白质,再利用酶联免疫分析实验进一步验证差异蛋白的变化。结果
1.针刺太冲与非经非穴对SHR血压的影响。组间比较结果：针刺前收缩压即原始收缩压的比较,太冲组、非经非穴组及模型组之间大鼠收缩压均无显著性差异(均P>0.05)。于针刺第1d,太冲组、非经非穴组及模型组之间大鼠收缩压均无显著性差异(均P>0.05)。于针刺第2d,与非经非穴组、模型组比较,太冲组大鼠收缩压显著降低(P<0.01),提示于针刺第2 d,太冲组大鼠收缩压开始降低。于针刺第3 d,与模型组比较,非经非穴组大鼠收缩压显著升高,提示针刺非经非穴可能是一种非良性刺激,导致大鼠收缩压显著升高。于针刺第4d,太冲组、非经非穴组及模型组之间大鼠收缩压均无显著性差异(均P>0.05)。于针刺第5d,与非经非穴组比较,太冲组大鼠收缩压显著降低(P<0.01)。于针刺第6、第7d,与非经非穴组、模型组比较,太冲组大鼠收缩压显著降低(均P<0.01),提示针刺太冲穴于第5d开始稳定的降压水平。与正常组比较,不同时间段太冲组、非经非穴组以及模型组大鼠收缩压均显著性升高(均P<0.01),提示本实验针刺太冲穴和非经非穴都未能使收缩压恢复至正常水平。
组内比较结果：与第1d比较,太冲组大鼠收缩压于第2d开始显著降低(P<0.01),然而于第3、第4d收缩压有小幅度升高(P>0.05),然后又于第5d开始降低(P<0.01),直到第6d、第7d收缩压降低水平保持稳定(均P<0.01),提示针刺太冲穴于第5d开始稳定的降压水平。非经非穴组于不同时间段收缩压均无显著性差异(P>0.05)。与第3d相比,模型组大鼠收缩压于第6d显著升高(P<0.01),与第4d比较,模型组大鼠收缩压于第6d和第7d均显著升高(均P<0.01),提示模型组大鼠的收缩压呈上升趋势。正常组于不同时间段收缩压均无显著性差异(P>0.05)。
2.各组大鼠延髓二维凝胶电泳图谱及其分析结果。本实验建立了正常SD大鼠、SHR以及对SHR针刺太冲与非经非穴后延髓的二维凝胶电泳图谱,共发现了SD大鼠延髓有621±30个蛋白点,SHR延髓有621±20个蛋白点,SHR经针刺太冲后延髓有621±40个蛋白点,SHR经针刺非经非穴后延髓有621±20个蛋白点,其中,与SHR组比较,SHR经针刺太冲后延髓表达有差异的蛋白点88个(大于2倍)；SHR经针刺非经非穴后延髓表达有差异的蛋白点91个(大于2倍),正常SD大鼠延髓表达有差异的蛋白点47个(大于2倍)。
3.鉴定出的差异蛋白质在各组大鼠延髓表达的结果。(1)与模型组比较,太冲组大鼠延髓下调的蛋白有5个(大于2倍)：突触素蛋白1,NAD依赖的脱乙酰酶,蛋白激酶C抑制蛋白,电子转移黄素蛋白和髓磷脂碱蛋白；上调的蛋白有6个(大于2倍)：谷氨酸脱氢酶,谷胱甘肽S转移酶,醛脱氢酶2, Rho GDP解离抑制因子,DJ-1蛋白和过氧化物歧化酶。(2)与模型组比较,非经非穴组大鼠延髓下调的蛋白有1个(大于2倍)：电子转移黄素蛋白；上调的蛋白有3个(大于2倍)：热休克蛋白90,泛素水解酶和DJ-1蛋白。(3)与非经非穴组比较,太冲组大鼠延髓下调的蛋白有7个(大于2倍)：热休克蛋白90,突触素蛋白1,丙酮酸激酶,NAD依赖的脱乙酰酶,蛋白激酶C抑制蛋白1,泛素水解酶和髓磷脂碱蛋白；上调的蛋白有6个(大于2倍)：谷氨酸脱氢酶,醛脱氢酶2,谷胱甘肽S转移酶,Rho GDP解离抑制因子,DJ-1蛋白及过氧化物歧化酶。(4)与正常组比较,模型组大鼠延髓下调的蛋白6个(大于2倍)：丙酮酸激酶,谷氨酸脱氢酶,谷胱甘肽S转移酶,醛脱氢酶2,泛素水解酶和过氧化物歧化酶；上调的蛋白有7个(大于2倍)：热休克蛋白90,突触素蛋白1,NAD依赖的脱乙酰酶,电子转移黄素蛋白,蛋白激酶C抑制蛋白1,蛋白激酶C抑制蛋白1和髓磷脂碱蛋白。(5)与正常组比较,太冲组大鼠延髓下调的蛋白有3个(大于2倍)：过氧化物歧化酶,过氧化物歧化酶和髓磷脂碱蛋白；上调的蛋白有4个(大于2倍)：热休克蛋白90, Rho GDP解离抑制因子,DJ-1蛋白和过氧化物歧化酶。(6)与正常组比较,非经非穴组大鼠延髓下调的有4个(大于2倍)：谷氨酸脱氢酶,醛脱氢酶2,谷胱甘肽S转移酶和过氧化物歧化酶；上调的蛋白有7个(大于2倍)：热休克蛋白90,突触素蛋白,NAD依赖的脱乙酰酶,蛋白激酶C抑制蛋白1,泛素水解酶,DJ-1蛋白和髓磷脂碱蛋白。
5.用酶联免疫分析结果。(1)各组大鼠延髓谷胱甘肽S转移酶表达的变化。模型组大鼠延髓谷胱甘肽S转移酶表达显著低于正常组(P<0.01)。经针刺治疗后,太冲组大鼠延髓谷胱甘肽S转移酶表达显著高于模型组、非经非穴组(均P<0.01),与正常组比较无统计学差异(P>0.01),而非经非穴组大鼠延髓谷胱甘肽S转移酶表达与模型组比较无统计学差异(P>0.01),且低于正常组(P<0.01)。(2)各组大鼠延髓醛脱氢酶表达的变化。模型组大鼠延髓醛脱氢酶表达显著低于正常组(P<0.01)。经针刺治疗后,太冲组大鼠延髓醛脱氢酶表达显著高于模型组、非经非穴组(均P<0.01),而非经非穴组大鼠延髓醛脱氢酶表达与模型组比较无统计学差异(P>0.01),且低于正常组(P<0.01)。(3)各组大鼠延髓蛋白激酶C表达的变化。模型组大鼠延髓蛋白激酶C表达与正常组无显著性差异(P>0.01)。经针刺治疗后,太冲组大鼠延髓蛋白激酶C表达无明显变化,与模型组、正常组、非经非穴组比较(均P>0.01)。谷胱甘肽S转移酶、醛脱氢酶的ELISA结果与二维凝胶电泳结果一致,蛋白激酶C的ELISA结果与二维凝胶电泳蛋白激酶C抑制蛋白1的结果不一致。
结论
1.针刺太冲穴可显著降低SHR高血压,但未能使SHR高血压恢复至正常水平。
2.针刺非经非穴降低SHR高血压效果不明显,甚至可能异常升高SHR高血压。
3.针刺太冲可下调SHR延髓内促进兴奋性神经递质释放、诱发炎症反应、氧化磷酸化及信号转导相关蛋白的表达,上调氨基酸氧化分子伴侣、抑制氧化应激及信号转导相关蛋白的变化,其作用机制是多途径、多环节、多靶点的综合效应。
4.针刺非经非穴可下调SHR延髓内能量代谢相关蛋白的表达,上调氨基酸氧化分子伴侣、蛋白质分解代谢以及诱发炎症反应相关蛋白的表达,其作用机制是较少途径、较少环节、较少靶点的简单效应。
5.经穴效应及其作用机制存在一定的特异性。
Objective
In order to research the effect of acupuncture at TaiChong point (LR3) and non-acupoint on the blood pressure in Spontaneously Hypertensive Rats (SHR), and investigate, in whole level of protein expression, the central nerve mechanism of acupuncturing TaiChong point (LR3) to treat SHR, which provide the experimental clue for searching the specific indicators of biological effects that connect to the meridian effect, two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technologies were used and the differentially expressed proteins in the medulla oblongata of SHR were also separated and identified.
Methods
309-week-old male SHR were randomly divided into model group, TaiChong group and non-acupoint group, each group involve 10 SHR. And another normal control group involves 10 SD rats of normal blood pressure with the same weight with SHR. In the TaiChong group, acupuncture bilateral Taichong points (located between the 1st and the 2nd metatarsal of dorsal hind); while in non-acupoint group acupuncture bilateral non-acupoint (located at the indentation between the 3rd and 4th metatarsal of dorsal hind). As rats awake, in Taichong group and non-acupoint group, daily acupuncture 5 minutes per times, twisting nedlle 80±5 times/min frequently, Twist nedlle 180±5 degrees in rang. In Taichong group, the needle towards Yongquang but not penetrate or reach the plantar skin, rather than in non-acupoint group, needle perpendicular to the skin and penetrate the skin.5 minutes after acupuncture, using BP-6 non-invasive blood pressure measuring system for tail arterial blood pressure measurement in rats. Acupuncture treatment continnues totally 7 weeks, all the rats brains were removed at the end, then separated in the ice for the medulla oblongata. And the medulla oblongata is cracked, to Sonication, desalted, seperated for proteins with the 2-DE. Geting the gel images with Typhoon9200 scanner, analyzing the differences between the various groups of protein with Imagesaster 6.0. With MALDI-TOF-TOF tandem MS analysis, protein mass image was obtained. And with the GPS software, to search and identify protein in the MASCOT database identification, then used enzyme-linked immunosorbent assay (ELISA) test to further verify the identification of the protein.
Results
1. The influence of acupuncture at Taichong point (LR3) or non-acupoints on the blood pressure SHR'blood pressure. (1) Comparison results among groups: The original, before acupuncture, systolic blood pressure of Taichong group, and the non-acupoint group and model group were not significantly different (all P> 0.05). The 1st day in acupuncture, in Taichong group, and the non-acupoint group and model group, systolic blood pressure between the rats were not significantly different (all P> 0.05). The next day in acupuncture, compared with the model group and non-acupoints group, systolic blood pressure of Taichong group significantly decreased (P<0.01), prompting that systolic blood pressure of Taichong group started to decrease in the next day in acupuncture. The 3rd day in acupuncture, compared with model group, systolic blood pressure of the non-acupoint group were significantly higher, suggesting that the acupuncture at non-acupoint may be a non-benign stimulus, resulting in a significantly increased of systolic blood pressure in rats. On the 4th in acupuncture, systolic blood pressure of Taichong group and the non-acupoint group and model group were not significantly different (all P> 0.05). On the 5th day of acupuncture, compared with non-acupoint, Taichong group's blood pressure decreased significantly (P<0.01). In acupuncture 6th, 7th day, compared with the model group and non-acupoints group, Taichong group' systolic blood pressure were significantly lower (P<0.01), suggested acupuncture at Taichong points begin to stabilize the blood pressure level in the 5th. In different time periods, in the model group, Taichong group and non-Meridian Group, systolic blood pressure were significantly higher than normal group (P<0.01), prompted acupuncture at Taichong or non-acupoint were not able to restore blood pressure to normal levels in this study.(2) Comparison results in each group: Compared with the 1st day, Taichong group systolic blood pressure decreased significantly in the next day (P<0.01); but on the 3rd and the 4th day, the systolic blood pressure were increased in small rate (P> 0.05), and then began to decrease at the 5th day (P<0.01), until the 6th day,7th day the reducing of systolic blood pressure remained stable (P<0.01), suggested acupuncture at Taichong points begin to stabilize the blood pressure level in the 5th. Non-acupoints group'systolic blood pressure were not significantly different in different time (P> 0.05). Compared with the 3rd day, the model rats'systolic blood pressure significantly increased on the 6th day (P<0.01); compared with the 4th day, model group's systolic blood pressure were significantly increased in the 6th day and the 7th days (P<0.01), prompting that systolic blood pressure of model group was rising. In different time, in nomal group, systolic blood pressure between the rats were not significantly different (all P> 0.05).
2. The two-dimensional gel electrophoresis (2-DE) image of rat's medulla in each group, and its analysis. In this study,2-DE image were successfully obtained for normal SD rats, SHR and the SHR after acupuncturing Taichong or non-acupoint; and 621±30 proteins in normal SD rats'medulla,621±20 proteins in SHR's medulla, 621±40 proteins in medulla of SHR after acupuncturing Taichong,621±20 proteins in medulla of SHR after acupuncturing non-acupoint were found also. Compared with SHR in model group,88 proteins were found differentially expressed in medulla from Taichong group (expression ratio of protein was beyond 2 folds); 91 proteins were found differentially expressed in medulla oblongata from non-acupoints group (expression ratio of protein was beyond 2 folds),47 proteins were found differentially expressed in medulla oblongata from normal group (expression ratio of protein was beyond 2 folds).
3. Differences in protein expression in the medulla of rats that dentified. (1) In medulla of Taichong group, compared with model group,5 proteins were down-regulated (expression ratio of protein was beyond 2 folds):Synapsin-1, NAD dependent deacetylase, electron transfer flavin protein and myelin basic protein; 6 proteins were up-regulated (expression ratio of protein was beyond 2 folds): glutamate dehydrogenase, glutathione S transferase, aldehyde dehydrogenase 2, Rho GDP dissociation inhibitor, DJ-1 protein and superoxide dismutase. (2) In medulla of non-acupoint group, compared with model group,1 proteins were up-regulated (expression ratio of protein was beyond 2 folds):electron transfer flavoprotein; 3 proteins were down-regulated (expression ratio of protein was beyond 2 folds): Heat shock protein 90, the Ubiquitin hydrolase isozyme and DJ-1 protein. (3)In medulla of Taichong group, compared with non-acupoints group,7 proteins were down-regulated (expression ratio of protein was beyond 2 folds):Heat shock protein 90, Synapsin-1, pyruvate kinase, NAD dependent deacetylases enzyme, protein kinase C inhibitor protein 1, Ubiquitin hydrolase isozymeand myelin basic protein; 6 proteins were up-regulated (expression ratio of protein was beyond 2 folds):glutamate dehydrogenase, aldehyde dehydrogenase 2, glutathione S transferase, Rho GDP dissociation inhibitor, DJ-1 protein and superoxide dismutase. (4) In medulla of model group, compared with normal group,6 proteins were down-regulated (expression ratio of protein was beyond 2 folds):pyruvate kinase, glutamate dehydrogenase, glutathione S transferase, aldehyde dehydrogenase 2, Ubiquitin hydrolase isozyme and superoxide dismutase; 7 proteins were up-regulated (expression ratio of protein was beyond 2 folds):Heat shock protein 90, Synapsin-11, NAD dependent deacetylase, electron transfer flavin protein, protein kinase C inhibitor protein 1, and myelin basic protein. (5) In medulla of Taichong group, compared with normal group,3 proteins were down-regulated (expression ratio of protein was beyond 2 folds):superoxide dismutase and myelin basic protein;
4 proteins were up-regulated (expression ratio of protein was beyond 2 folds): Heat shock protein 90, Rho GDP dissociation inhibitor, DJ-1 protein and superoxide dismutase. (6) In medulla of non-acupoint group, compared with the normal group, 4 proteins were down-regulated (expression ratio of protein was beyond 2 folds): glutamate dehydrogenase, aldehyde dehydrogenase 2, glutathione S transferase and over superoxide dismutase; 7 proteins were up-regulated (expression ratio of protein was beyond 2 folds):Heat shock protein 90, synapsin 1, NAD dependent deacetylase, protein kinase C inhibitor protein 1, Ubiquitin hydrolase isozyme, DJ-1 protein and myelin basic protein.
5. results of ELISA
(1) The expression changes of glutathione S transferase in medullary among each group. In the medulla model group, compared with normal group, the expression of glutathione S transferase was significantly down-regulated (P<0.01). After acupuncture treatment, the expression of glutathione S transferase in the medulla of Taichong group were significantly up-regulated, compared with the model group, and the non-acupoint group, (P<0.01); but the expression of glutathione S transferase in the medulla of non-acupoint group has no significant difference form the model group (P> 0.01), and lower than the normal group (P<0.01). (2) The expression changes of aldehyde dehydrogenase in the medulla among each group. In the medulla model group, compared with normal group, the expression of aldehyde dehydrogenase was significantly down-regulated (P<0.01). After acupuncture treatment, the expression of aldehyde dehydrogenase in the medulla of Taichong group were significantly up-regulated, compared with the model group, and the non-acupoint group, (P<0.01); but the expression of aldehyde dehydrogenase in the medulla of non-acupoint group has no significant difference form the model group (P> 0.01), and lower than the normal group (P<0.01) (3) The expression changes of protein kinase C in medullary among each group. The expression of protein kinase C in the medulla of model group has no significant difference form the nomal group (P> 0.01); After acupuncture treatment, expression of protein kinase C in medullary of Taichong group has no significant changes from the model group, normal group, and the non-acupoint group (all P> 0.01). ELISA results of Glutathione S transferase, aldehyde dehydrogenase are consistent with the two-dimensional gel electrophoresis; meanwhile the ELISA results of protein kinase C are inconsistent with the results of two-dimensional gel electrophoresis of protein kinase C inhibitor protein 1.
Conclusion
1. Acupuncture Taichong can reduce the hypertension of SHR, but were not able to restore blood pressure to normal levels in this study.
2. acupuncture at non-acupoints didn't have an obvious effect on decreasing the blood pressure in SHR, even cause increase in SHR hypertension.
3. Acupuncture Taichong Point can down-regulated expressions of the proteins that promote excitatory neurotransmitter, induced inflammatory response, oxidative phosphorylation and induce oxidative stress protein, and up-regulate amino acid oxidation chaperones and energy metabolism and oxidative stress in SHR' medulla. Its mechanism is a multi-channel, multi-link, the combined effect of multiple targets.
4. Acupuncture non-acupoint can down-regulated expressions of the proteins that promote energy metabolism, and up-regulate amino acid oxidation, protein catabolism and expressions of protein that induced inflammatory response in medullary of SHR. The mechanism is less means, fewer links, fewer target Point of simple effect.
5. Acupoints effect and its mechanism has some specificity.
引文
[1]王吉耀.内科学.北京：人民卫生出版社,2005.
[2]刘明波,王文.中国心血管病报告2008-2009.北京：中国大百科全书出版社,2009.
[3]中华人民共和国卫生部.2008中国卫生统计年鉴.北京：中国协和医科大学出版社,2008.
[4]Kearney P M, Wthelton M, Reynolds K, et al. Global burden of hypertension:analysis of worldwide data. Lancet,2005,65(9455):217-223.
[5]William J Elliott, Peter M Meyer. Incident diabetes in clinical trials of antihypertensive drugs:a network meta-analysis. Lancet,2007,369 (9557):201-207
[6]Chobanian A V, Bakris G L, Black H R, et al. Seventh report of the Joint National Committee on the Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7). Hypertension,2003,42(6): 1206-1252.
[7]张廷杰,伍悦蕾.从欧洲2007ESH/ESC高血压指南看高血压防治趋势.中华老年多器官疾病杂志,2009,8(5)：477-80.
[8]郭明冬,崔玲,周文泉.周文泉教授从肝论治原发性高血压经验.中国临床康复,2006,10(19)：168-169.
[9]吴焕林.邓铁涛教授治疗高血压病临床经验辑要.河南中医,2005,25(5)：16-17.
[10]屈会化,赵琰,曲荣波,等.自发性高血压大鼠早期中医证候的研究.中医杂志,2008,49(5)：446-448.
[11]申鹏飞,石学敏.动态血压监测评价“活血散风,平肝降逆”针刺治疗原发性高血压病的临床疗效分析.世界传统医药国际科技大会论文集(针灸部分),2009：111-113.
[12]刘建永,杨佃会,白田雨.针灸治疗原发性高血压临床用穴频次分析.山东中医药大学学报,2009,33(3)：215-216.
[13]Harun Ozer OU,Sait A, Hakan S, et al. Possible Electrical Stimulation on Rostral Ventrolateral Medulla That Causes Significant Hypertensive and Tachycardic Changes during Endovascular Treatment of a PICA Aneurysm Abutting the Left Medullary Sulcus. AJ of Neuroradiology,2005,26(4):522-524.
[14]Cobos A, Lima D, Almeida A, et al. Brain afferents to the lateral caudal ventrolateral medulla:A retrograde and anterograde tracing study in the rat. Neuroscience,2003,120(2):485-498
[15]Kantzides A, Simmmons E. Nos-containing neurons in the hypothalamus and medulla project to the RVLM. Brain Res,2005,1037(2):25-34.
[16]Saha S, Drinkhill M J, Moore J P, et al. Central nucleus of amgydala projections to rostral ventrolateral medulla neurones activated by decreased blood pressure. Neurosci,2005,21(7):1921-1930.
[17]Gaykema R P, Chen C C, Goehler L E. Organization of immune-responsive medullary projections to the bed nucleus of the stria terminalis, central amygdala and paraventricular nucleus of the hypothalamus:evidence for parallel viscerosensory pathways in the rat brain. Brain Res,2007,1130(1):130-145.
[18]张春沿,郝强,张宇鹏,等.延髓腹外侧区的神经通路及其功能.河北师范大学学报(自然科学版),2008,32(5)：668-672.
[19]百晓生.延髓心血管中枢.http://baike. baidu. com/view/1715773. htm,2010-04-20.
[20]Dampney R A. Blood pressure regulation by C1 neurons in the rostral ventrolateral medulla:new light on the subject. Physiology,2009,587(23): 5513-5518
[21]Wang W Z, Yuan W J, Pan Y X, et al. Non-NMDA receptors within caudal ventrolateral medulla are nivolved in transmission of baroreflex of rats. Acta Pharmacol Sin,2003,24(8):783-789.
[22]Miyashita T, Wiliiams C L. Glutamatergic transmission in the nuclers of the solitary tract modulates memory through influences on amygdala noradrenergic systems. Behav Neurosci,2002,116(1):13-21.
[23]周苏娅,朱一平,高峰,等.腓深神经对最后区心血管中枢活动的调节作用.中国应用生理学杂志,2003,19(1)：7-8.
[24]Xia C M, Chen J, Wang J,.et al. Differential expressions of nNOS and iNOS in the rostral ventrolateral medulla induced by electroacupuncture in acute myocardial ischemia rats. Acta Physiologica Sinica,2008,60(4): 453-461.
[25]周逸平,王月兰,方志斌.针刺对SHR血压及NE、DA、5-HT含量的影响和血压与血粘度的关系.针刺研究,1995,20(3)：55-62.
[26]刘健华,李江山,严洁,等.针刺“足三里”对大鼠孤束核P物质的影响.针刺研究,2003,28(1)：5-9
[27]张进,陈东风,李伊为,等.电针内关穴诱导大鼠延髓原癌基因c-fos表达.广州中医药大学学报,2001,18(2)：134-136
[28]Swinbanks D. Government backs proteome protosal. Nature,1995,378 (6558): 386-653.
[29]钱小红,贺福初.蛋白质组学理论与方法.北京：科学出版社,2005：1-9.
[30]刘建君,白克镇.精神分裂症的蛋白组学研究新进展.神经疾病与精神卫生.2008,8(2)：153-155.
[31]Panday A, Mann M. Proteomics to study genes and genomes. Nature, 2000,405 (6788):837-846.
[32]Anne-Claude Gavin, et al. Functional organization of the yeast proteome by systematic analysis of protein complexes. Nature,2002,415(6868):141-147.
[33]Service, Robert F. Proteomics:A Proteomics Upstart Tries to Outrun the Competition. Science,2001,294 (5549):2079-2080.
[34]Gerstein, Mark, Lan, Ning, Jansen, Ronald. Enhanced:Integrating Interactions. Science,2002,295 (5553):284-287.
[35]Kuramitsu Y, Nakamura K. Proteomic analysis of cancer tissues:shedding light on carcinogensesis and possible biomarker. Proteomics,2006,20(6): 5650-5661
[36]Bertucci F, Concalves A. Clinical proteomics and breast cancer:astategise for diagnostic and therapeutic biomarker discovery. Future Oncol,2008,4(2):271-287.
[37]Almeras L, Lefras D, Drobecq H, et al. New antigenic candidates analysis. Proteomics,2004,4(7):2184-2194.
[38]Labugger R, Organ L, Collier C, et al. Extensive troponin I and T modification dectected in serum from patients with acute myocardial infarction. Circulation,2000,102(11):1221-1226.
[39]Mei J, Kolbin D, Kao H T, et al. Proterin expression profiling of postmortem brain in schizophrenia. Schizophr Res,2006,84(1):204-213.
[40]Butterfield D A, Boyd kinball D, Castegna A. Proteomics in Alzheimer's of neuro-Degeneration. Neurochemi,2003,86(6):1313-1327.
[41]李志刚,耿昊,苑晓晨,等.督脉电针对急性脊髓损伤6h后大鼠差异蛋白质组学的机理研究.北京中医药大学学报,2008,31(3)：202-226.
[42]温锦荣,赵晓峰,王舒,等.局灶性脑缺血大鼠海马蛋白质组差异表达图谱的构建及针刺影响.天津中医药,2008,25(2)：96-99.
[43]苑晓晨,宋金玲,田贵华,等.不同时段电针对急性脊髓损伤大鼠作用机制的蛋 白质组学分析.针刺研究,2009,34(2)：75-82.
[44]李忠仁.实验针灸学.北京：中国中医药出版社,第1版,2007：255-257
[45]Kalish L A, Buczynski B, Connell P, et al. Stop hypertension with the acupunture research program(SHARP):clinical trial design and screening results. Conrtolled Clinical Trials,2004,25(1):76-103.
[46]杨路,赖新生.针刺十二原穴对高、低血压大鼠模型血压影响的经穴效应规律研究.广州中医药大学博士论文,2008：68-69.
[48]Paxinos G, Watson C. The rat brain in stereotaxic coordinates. USA:Academic Press Inc, the sixth edition,2007:25-807.
[49]姚一康,杨伟敏,严国锋.对SHR大鼠病理特性测定研究.上海实验动物科学,2003,23(2)：75-82.
[50]宿中笑,肖达,睢久红,等.电针对自发性高血压大鼠血压和主动脉血管重构的影响.上海针灸杂志,2003,22(5)：20-24.
[51]杨华元,钟小红,刘堂义等.针刺仿真手法对高血压大鼠血压及心肌血管紧张素Ⅱ的影响.针刺研究,2008,33(3)：1186-1190.
[52]吴焕林,李晓庆,王侠.针刺太冲穴对65例肝阳上亢型高血压病患者的即时降压效应.中医杂志,2008,49(7)：622-624.
[53]尹艳茹.L-精氨酸对应激性大鼠血压的影响.中国临床药理学与治疗学,2002,7(6)：529-531.
[54]李漫松.侧脑室内注射BGN对应激性高血压大鼠心血管反应的影响.吉林大学学报(医学版),2003,29(4)：389-392.
[55]Kunz M, Scholl KE, Schu U, et al. GABAergic modulation of diffuse noxious inhibitory controls (DNIC):a test by use of lorazepam. Experimental Brain Research,2006,175(2):.363-371.
[56]王家有,唐纯志,赖新生,等.太冲穴中等强度针刺可降低高血压大鼠血压及血浆内皮素含量.针刺研究,2010,待刊.
[57]Tam K C, Yiu H H. The effect of acupuncture on essential hypertension. Am J Chin Med,1975,3(4):369-379.
[58]吴远华,朱广旗,林兴友,等.针刺曲池、太冲对高血压病患者血中ET和ACE的影响及疗效探讨.中国中西医结合杂志,2004,24(12)：1080-1083.
[59]Flachskampf, F A, Joachim Gallasch, Olaf Gefeller, et al. Randomized trial of acupuncture to lower blood pressure. Circulation,2007,115(24):3121-3129.
[60]Eric A. Macklin, Peter M.Wayne, Leslie A, et al. Stop Hypertension with the Acupuncture Research Program (SHARP):results of a randomized, controlled clinical trial[J]. Hypertension,2006.48(5):834-845.
[61]陈邦国,钱春艳,张静宁,等.针刺风池穴对高血压病降压疗效临床观察.上海针灸杂志,2006,25(3)：15-18.
[62]王侠,吴焕林,李晓庆.针刺太冲穴对动态血压的降压疗效观察.新中医,2007,39(11)：21-22.
[63]吴根诚,曹小定.针刺疗法走向世界的历史与影响因素即几点思考.中西医结合学报,2003,1(4)：247-251.
[64]Yin C, Seo B, Park HJ, et al. Acupuncture, a promising adjunctive therapy for essential hypertension:a double-blind, randomized, controlled trial. Neurol Res,2007,29 Suppl 1:98-104.
[65]李吉梅.针刺为主治疗顽固性高血压25例.河南中医,2005,25(1)：65-67.
[66]黄凡,姚国新,黄小丽,等.针刺治疗痰淤阻络型高血压病临床观察.中国针灸,2007,27(6)：403-406.
[67]王葳,李坤成,单保慈,等.针刺太冲穴的脑功能MRI研究.中华放射学杂志,2006,40(1)：29-35.
[68]Berman B M, Lao L, Langen berg P, et al. Effectiveness of acupuncture as adjunctive therapy in osteoarthritis of the knee:a randomized, controlled trial. Ann Intern Med,2004,141(12):901-910.
[69]Witt C,Brinkhaus B, Jena S, et al. Acupuncture in patients with ostearthritis of the knee:a randomised trial. Lancet,2005,366(9480): 136-143.
[70]艾林,戴建平,赵百孝,等.针刺镇痛机制的功能磁共振成像研究.中国医学影像技术,2004,20(8)：1197-1200.
[71]田小平,李瑛,梁繁荣,等.经穴与非经穴效应比较研究的现状与思考.中国针灸,2008,28(12)：907-909.
[72]胡卡明,王承平,谢慧君,等针刺穴位与非穴位激活不同脑区的效应观察.中国针灸,2006,26(3)：205-207.
[73]Maioli C, Falciati L, Marangon M, et al. Short-and long-term modulation of upper limb motor-evoked potentials induced by acupuncure. Eur J Neurosci,2006,23(1):1931-1938.
[74]李瑛,梁繁荣,郑晖,等.针刺经穴与非经非穴治疗偏头痛急性发作期的随机对照研究.世界传统医药国际科技大会论文集(针灸部分),2009：114-119
[75]Zhang W, LI JL, Hosaka M, et al. Cyclosporine A-induced hypertension involves synapsin in renal sensory nerve endings. Proc Natl Acad Sci USA,2000,97(17):9765-9770
[76]Koji Ito, Yoshitaka Hirooka, Nobuaki Hori, et al. Inhibition of Rho-Kinase in the nucleus tractus solitarius enhances glutamate sensitivity in rats. Hypertension online,2005, DOI:10.1161/01.
[77]张明亮,严志强,齐颖新,等.Rho蛋白解离抑制因子α在高血压大鼠胸主动脉的表达.解剖学报,2009,40(4)：625-629
[78]Wiedemann B, Franke W. Identification and localization of synaptophysin, an integral membrane glycoprotein of Mr 38,000 characteristic of presynapticvesicles.Cell,1985,41 (3):1017-1029
[79]Petra K, Heinrich B, Hubert R. Calcium-dependent serine phosphorylation of synaptophysin. Journal of Neur.2006,47(4):1302-1304.
[80]赵合庆,刘晶,余锋,等.经颅磁刺激对短暂性大脑中动脉闭塞大鼠突触素表达和超微结构改变的影响.中华脑血管并杂志(电子版),2008,2(1)：17-22
[81]沈丽,买鸿宴,南燕,等.自发性高血压大鼠中缝背核的超微结构.高血压杂志,1998,6(2)：209-213
[82]Evergren E, Benfenati F, Shupliakov O.The synapsin cycle:A view frm the synaptic endocytic zone. Neuroscience Reseach,2007,85(12):2648-2656.
[83]Lonart G, Simsek-Duran F. Deletion of synapsins I and II genes alters the size of vesicular pools and rabphilin phosphorylation. Bran Res,2006 1107:42-51
[84]Di Fraia R, Wilquet V, Ciardiello M A, et al.NADP+ dependent glutamate dehydrogenase in the antarctic psychrotolerant bacterium psychrobacter sp. Tadl. characterization, protein and DNA sequence and relationship to other glutamate dehydrogenases. Biochem,2000,267(1):121-131
[85]付海英,张纪周,杨宇丹,等.海洛因对大鼠谷氨酸脱氢酶基因表达的影响.中国药物依赖性杂志,2006,15(6)：440-444
[86]Cansein R O, Bonagamba L G, Machado B H. Bradycardic and hypotensive responses to microinjection of L-glutamate into the lateral aspect of the commissural NTS are blocked by an NMDA receptor antagonist. Bran Res,2000,852(1):68-75
[87]孟培燕,孙国杰,刘胜洪,等.电针预处理对血管性痴呆大鼠神经细胞谷氨酸-NMDA受体转导通路的影响.针刺研究,2008,33(2)：236-239
[88]崔毅军,汪石右.针刺对膀胱机能亢进大鼠蓝斑谷氨酸影响的动态研究.上海中医药大学博士学位论文,2005：2
[89]Rhian M. Touyz. Reactive Oxygen species, vascular oxidative stress,and redox signaling in hypertion. Hypertension,2004,44 (2):248-256
[90]Lie G, Wang W Z, Wang W, et al. Effects of angiotensin type 2 receptor overexpression in the rostral ventrolateral medulla on blood pressure and urine excretion in normal rats. Hypertension,2008,51 (2):521-527
[91]Leenen F H. Brain mechanisms contributing to sympathetic hyperactivity and heart failure. Circulation Res,2007,101 (3):221-223
[92]Ying W.NAD+/NADH and NADP+/NADPH in cellular functions and cell death:regulation and biological consequences. Antioxid Redox Signal, 2007,10 (2):179-206
[93]Infanger D W, Sharma R V, Davisson RL. NADPH oxidases of the brain:distribution, regulation and function. Axtioxid Redox Signal, 2006,8(2):1583-1596
[94]Freya Q. Schafer, Garry R, et al. Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radical Biology and Medicine,2001,30(11):1191-1212.
[95]周四桂,潘雪刁,唐富天,等.依那普利对肾性与自发性高血压大鼠左心室Enol及GSTM2表达的影响.中国药理学通报,2008,24(6)：736-740
[96]Iwai N, Tago N, Yasui N, et al. Analysis of 22 candidate genes for hypertension in the Japanese population. Hypertension,2004,22 (6):1119-1126.
[97]Custodis F, Eberl M, Kilter H, et al. Association of RhoGDIa with Racl GTPase mediates free radial production during myocardial hypertrophy. Cardiovasc Res,2006,71(2):342-351
[98]Pacaud P, Sauzeau V, Loirand G. Rho proteins and vascular diseases.Arch Mal Coeur Vaiss,2005,98(3):249-254
[99]Riha G M, Lin P H, Lumsden L B, et al. Roles of hemodynamic forces in vascular cell differentiation. Ann Biomech Eng,2005,33(6):772-779
[100]Pockley A G, Wu R, Lemne C, et al. Circulating heat shock protein 60 is associated with early cardiovascular disease. Hypertension,2000,36 (1): 303-307
[101]Amamoto K, Okamura T, Tamaki S, et al. Epidemiologic study of the association of low-Kmmitochondrial acetaldehyde dehydrogenase genotypes with blood pressure level and the prevalence of hypertension in a general population. Hypertension Res,2002,25(6):857-864
[102]Boggs J M. Myelin basic protein:a multifunctional protein. Cell Mol Life Sci,2006,63(1):1945-1961
[103]Maria Carolina Gongora, David G Harrison. Sad Heart from no SOD. Hypertension,2008,51:28-32
[104]Usmani S Z, Bona R, Li Z.17 AAG for HSP90 inhibition in cancer-from bench to bedside.Curr Mol Med,2009,9(5):654-664
[105]Moser C, Lang S A, Stoeltzing 0. Heat shock protein 90(Hsp90)as a molecular target for therapy of gastrointestinal cancer. Anticancer Res,2009,29(6):2031-2042
[106]Reikvam H, Ersvaer E, Bruserud 0. Heat shock protein 90 a potential target in the treatment of human acute myelogenous leukemia. Curr Cancer Drug Targets,2009,9(6):761-776
[107]Tsutsumi S, Beebe K, Neckers L. Impact of heat-shock protein 90 on cancer metastasis. Future Oncol,2009,5 (5):679-688
[108]Moleda L, Jurzik L, Froh M, et al.Role of HSP-90 for increased nNOS-mediated vasodilation in mesenteric arteries in portal hypertension. World J Gastroenterol,2010,16(15):1837-1844
[109]Moiseeva 0 M, Novoselov S S, Liasnikova E A, et al.Heat shock protein 70 as a marker of lesions to target organs in hypertension.Vestn Ross Akad Med Nauk,2004,1(1):6-9
[110]Wright B H, Corton J M, E1-Nahas A M, et al. Elevated levels of circulating heat shock protein 70 in peripheral and renal vascular disease. Heart Vessels,2000,15(1):18-22
[111]Pockley A G, De Faire U, Kiessling R, et al. Circulating heat shock protein and heat shock protein antibody levels in established hypertension. Hypertension,2002,20 (1):1815-1829
[112]Moon K H, Kim B J, Song B J.Inhibition of mitochondrial aldehyde dehydro-genase by nitric oxide-mediated S-nitrosylation. Febs Lett,2005,579(27):6115-6120
[113]Daiber A, Wenzel P, Oelze M, et al. Mitochondrial aldehyde dehydrogenase (ALDH-2)-marker of and marker for nitrate tolerance in response to nitroglycer in treatment. Chem Biol Int,2009,178(1-3):40-47
[114]Chen Z Q, Stamler J S. Bioactivaction of nitroglycerin by the mitochondrial aldehyde dehydrogenase. Trends Cardiovasc Med,2006,16(8):259-265
[115]Sun A J, Wang K Q, Ge J B, et al. Proteomic analysis of myocardial mitochondria in rats with heart failure. Cardiovasc Drugs Ther,2004,18(1):21-29
[116]Hui P, Nakayama T, Morita A, et al. Common single nucleotide polymorphisms in Japanese patients with essential hypertension:aldehyde dehydrogenase 2 gene as a risk factor independent of alcohol consumption. Hypertension Res,2007,30(7):585-592
[117]韩济生.神经科学原理[M].第2版,北京：北京医科大学出版社,1999：1050-1059
[118]乔松,冯加纯,杨晶,等.高血压大鼠脑内小血管改变及其继发损伤的研究.中风与神经疾病杂志,2006,23(3)：290-294
[119]宿宝贵,潘三强,,王家有,等.慢性肾性高血压大鼠脑组织磁共振成像观察和超微结构的研究.解剖学报,2005,35(6)：636-639
[120]Constantinescu C S,Goodman D B,Hilliard B, ea al.Murine macrophages stimulated with central and peripheral nervous system myelin or purified myelin proteins release inflammatory products. Neurosci Lett.2000,287 (3):171-174
[121]刘红岩,杨贵贞.髓磷脂碱性蛋白促进单个核细胞对内皮细胞的粘附效应.中国免疫学杂志,2000,16(8)：428-431
[122]Hidefumi Waki,Beihui Liu, Massao Miyake,et al. Junctional adhesion molecule-1 is upregulated in spontaneously hypertensive rats. Hypertension, 2007,49:1321-1327
[1]严季澜.建国40年来高血压病的针灸推拿治疗概况[J].天津中医,1990,(6)：37.
[2]佘延芬,于文,夏茜,等.近10年针灸治疗高血压病的研究进展[J].河北中医药学报,2000,15(3)：36.
[3]刘锦,乔嘉斌.针刺治疗高血压的研究进展[J].吉林中医药,2008,28(2)：152.
[4]Tam K C, Yiu H H. The effect of acupuncture on essential hypertension[J]. Am J Chin Med,1975,3(4):369.
[5]陈邦国,钱春艳,张静宁,等.针刺风池穴对高血压病降压疗效临床观察[J].上海针灸杂志,2006,25(3)：15.
[6]卫彦,孙忠人,寇吉友,等.针刺人迎穴治疗高血压病120例临床观察[J].针灸临床杂志,2006,22(2)：4.
[7]万文俊,马朝阳,熊修安,等.电针曲池穴治疗原发性高血压病疗效观察及机制探讨.中国针灸,2009,29(5)：349.
[8]Yin C, Seo B, Park HJ,et al. Acupuncture, a promising adjunctive therapy for essential hypertension:a double-blind, randomized, controlled trial[J]. Neurol Res,2007,29 Suppl 1:98.
[9]李吉梅.针刺为主治疗顽固性高血压25例[J].河南中医,2005,25(1)：65.
[10]黄凡,姚国新,黄小丽,等.针刺治疗痰淤阻络型高血压病临床观察.中国针灸,2007,27(6)：403.
[11]吴远华,朱广旗,林兴友,等.针刺曲池、太冲对高血压病患者血中ET和ACE的影响及疗效探讨.中国中西医结合杂志,2004,24(12)：1080.
[12]Flachskampf, F A, Joachim Gallasch, Olaf Gefeller, et al. Randomized trial of acupuncture to lower blood pressure[J]. Circulation,2007,115(24):3121.
[13]郭玉红.针刺治疗高血压病胰岛素抵抗的临床观察[J].中医药学报,2007,35(6)：51.
[14]Sugioka, K, Mao W, Woods J, et al. An unsuccessful attempt to treat hypertension with acupuncture[J]. Am J Chin Med (Gard City N Y),1977,5(1):39.
[15]Eric A. Macklin, Peter M.Wayne, Leslie A, et al. Stop Hypertension with the Acupuncture Research Program (SHARP):results of a randomized, controlled clinical trial[J]. Hypertension,2006.48(5):838.
[16]Lee H, Kim S Y, Park J, et al. Acupuncture for lowering blood pressure: systematic review and meta-analysis[J]. Am J Hypertens,2009,22:122.
[17]吴焕林.邓铁涛教授治疗高血压病临床经验辑要[J].河南中医,2005,25(5)：16.
[18]王侠,邹旭,李琼,等.针刺太冲穴治疗高血压的临床研究[J].实用医学杂志,2003,19(5)：565.
[19]朱广旗,吴远华,吴邦启,等.针刺曲池和太冲对高血压病不同证型的疗效[J].浙江中西医结合杂志,2006,16(1)：4.
[20]穆晓红,李巧霞.针刺太冲和足三里治疗高血压病临床观察[J].上海针灸杂志,2009,28(6)：328.
[21]张为.针刺悬钟穴治疗高血压病40例[J].河北中医,2006,28(2)：154.
[22]曹玲,侯广云,范文双.针刺颈部夹脊穴治疗颈源性高血压的疗效分析[J].针刺研究,2007,32(3)：195.
[23]吴焕林,李晓庆,王侠.针刺太冲穴对65例肝阳上亢型高血压病患者的即时降压效应[J].中医杂志,2008,49(7)：622.
[24]冯国湘,吴清明.针刺开“四关”加百会穴温针灸治疗原发性高血压的临床研究[J].中国针灸,2003,23(4)：193.
[25]刘建永,杨佃会,白田雨.针灸治疗原发性高血压临床用穴频次分析[J].山东中医药大学学报,2009,33(3)：215.
[26]杨路,赖新生.针刺十二原穴对高、低血压大鼠模型血压影响的经穴效应规律研究[D].广州中医药大学博士论文,2008：68
[2]刘明波,王文.中国心血管病报告2008-2009.北京：中国大百科全书出版社,2009.
[3]中华人民共和国卫生部.2008中国卫生统计年鉴.北京：中国协和医科大学出版社,2008.
[4]Kearney P M, Wthelton M, Reynolds K, et al. Global burden of hypertension:analysis of worldwide data. Lancet,2005,65(9455):217-223.
[5]William J Elliott, Peter M Meyer. Incident diabetes in clinical trials of antihypertensive drugs:a network meta-analysis. Lancet,2007,369 (9557):201-207
[6]Chobanian A V, Bakris G L, Black H R, et al. Seventh report of the Joint National Committee on the Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7). Hypertension,2003,42(6): 1206-1252.
[7]张廷杰,伍悦蕾.从欧洲2007ESH/ESC高血压指南看高血压防治趋势.中华老年多器官疾病杂志,2009,8(5)：477-80.
[8]郭明冬,崔玲,周文泉.周文泉教授从肝论治原发性高血压经验.中国临床康复,2006,10(19)：168-169.
[9]吴焕林.邓铁涛教授治疗高血压病临床经验辑要.河南中医,2005,25(5)：16-17.
[10]屈会化,赵琰,曲荣波,等.自发性高血压大鼠早期中医证候的研究.中医杂志,2008,49(5)：446-448.
[11]申鹏飞,石学敏.动态血压监测评价“活血散风,平肝降逆”针刺治疗原发性高血压病的临床疗效分析.世界传统医药国际科技大会论文集(针灸部分),2009：111-113.
[12]刘建永,杨佃会,白田雨.针灸治疗原发性高血压临床用穴频次分析.山东中医药大学学报,2009,33(3)：215-216.
[13]Harun Ozer OU,Sait A, Hakan S, et al. Possible Electrical Stimulation on Rostral Ventrolateral Medulla That Causes Significant Hypertensive and Tachycardic Changes during Endovascular Treatment of a PICA Aneurysm Abutting the Left Medullary Sulcus. AJ of Neuroradiology,2005,26(4):522-524.
[14]Cobos A, Lima D, Almeida A, et al. Brain afferents to the lateral caudal ventrolateral medulla:A retrograde and anterograde tracing study in the rat. Neuroscience,2003,120(2):485-498
[15]Kantzides A, Simmmons E. Nos-containing neurons in the hypothalamus and medulla project to the RVLM. Brain Res,2005,1037(2):25-34.
[16]Saha S, Drinkhill M J, Moore J P, et al. Central nucleus of amgydala projections to rostral ventrolateral medulla neurones activated by decreased blood pressure. Neurosci,2005,21(7):1921-1930.
[17]Gaykema R P, Chen C C, Goehler L E. Organization of immune-responsive medullary projections to the bed nucleus of the stria terminalis, central amygdala and paraventricular nucleus of the hypothalamus:evidence for parallel viscerosensory pathways in the rat brain. Brain Res,2007,1130(1):130-145.
[18]张春沿,郝强,张宇鹏,等.延髓腹外侧区的神经通路及其功能.河北师范大学学报(自然科学版),2008,32(5)：668-672.
[19]百晓生.延髓心血管中枢.http://baike. baidu. com/view/1715773. htm,2010-04-20.
[20]Dampney R A. Blood pressure regulation by C1 neurons in the rostral ventrolateral medulla:new light on the subject. Physiology,2009,587(23): 5513-5518
[21]Wang W Z, Yuan W J, Pan Y X, et al. Non-NMDA receptors within caudal ventrolateral medulla are nivolved in transmission of baroreflex of rats. Acta Pharmacol Sin,2003,24(8):783-789.
[22]Miyashita T, Wiliiams C L. Glutamatergic transmission in the nuclers of the solitary tract modulates memory through influences on amygdala noradrenergic systems. Behav Neurosci,2002,116(1):13-21.
[23]周苏娅,朱一平,高峰,等.腓深神经对最后区心血管中枢活动的调节作用.中国应用生理学杂志,2003,19(1)：7-8.
[24]Xia C M, Chen J, Wang J,.et al. Differential expressions of nNOS and iNOS in the rostral ventrolateral medulla induced by electroacupuncture in acute myocardial ischemia rats. Acta Physiologica Sinica,2008,60(4): 453-461.
[25]周逸平,王月兰,方志斌.针刺对SHR血压及NE、DA、5-HT含量的影响和血压与血粘度的关系.针刺研究,1995,20(3)：55-62.
[26]刘健华,李江山,严洁,等.针刺“足三里”对大鼠孤束核P物质的影响.针刺研究,2003,28(1)：5-9
[27]张进,陈东风,李伊为,等.电针内关穴诱导大鼠延髓原癌基因c-fos表达.广州中医药大学学报,2001,18(2)：134-136
[28]Swinbanks D. Government backs proteome protosal. Nature,1995,378 (6558): 386-653.
[29]钱小红,贺福初.蛋白质组学理论与方法.北京：科学出版社,2005：1-9.
[30]刘建君,白克镇.精神分裂症的蛋白组学研究新进展.神经疾病与精神卫生.2008,8(2)：153-155.
[31]Panday A, Mann M. Proteomics to study genes and genomes. Nature, 2000,405 (6788):837-846.
[32]Anne-Claude Gavin, et al. Functional organization of the yeast proteome by systematic analysis of protein complexes. Nature,2002,415(6868):141-147.
[33]Service, Robert F. Proteomics:A Proteomics Upstart Tries to Outrun the Competition. Science,2001,294 (5549):2079-2080.
[34]Gerstein, Mark, Lan, Ning, Jansen, Ronald. Enhanced:Integrating Interactions. Science,2002,295 (5553):284-287.
[35]Kuramitsu Y, Nakamura K. Proteomic analysis of cancer tissues:shedding light on carcinogensesis and possible biomarker. Proteomics,2006,20(6): 5650-5661
[36]Bertucci F, Concalves A. Clinical proteomics and breast cancer:astategise for diagnostic and therapeutic biomarker discovery. Future Oncol,2008,4(2):271-287.
[37]Almeras L, Lefras D, Drobecq H, et al. New antigenic candidates analysis. Proteomics,2004,4(7):2184-2194.
[38]Labugger R, Organ L, Collier C, et al. Extensive troponin I and T modification dectected in serum from patients with acute myocardial infarction. Circulation,2000,102(11):1221-1226.
[39]Mei J, Kolbin D, Kao H T, et al. Proterin expression profiling of postmortem brain in schizophrenia. Schizophr Res,2006,84(1):204-213.
[40]Butterfield D A, Boyd kinball D, Castegna A. Proteomics in Alzheimer's of neuro-Degeneration. Neurochemi,2003,86(6):1313-1327.
[41]李志刚,耿昊,苑晓晨,等.督脉电针对急性脊髓损伤6h后大鼠差异蛋白质组学的机理研究.北京中医药大学学报,2008,31(3)：202-226.
[42]温锦荣,赵晓峰,王舒,等.局灶性脑缺血大鼠海马蛋白质组差异表达图谱的构建及针刺影响.天津中医药,2008,25(2)：96-99.
[43]苑晓晨,宋金玲,田贵华,等.不同时段电针对急性脊髓损伤大鼠作用机制的蛋 白质组学分析.针刺研究,2009,34(2)：75-82.
[44]李忠仁.实验针灸学.北京：中国中医药出版社,第1版,2007：255-257
[45]Kalish L A, Buczynski B, Connell P, et al. Stop hypertension with the acupunture research program(SHARP):clinical trial design and screening results. Conrtolled Clinical Trials,2004,25(1):76-103.
[46]杨路,赖新生.针刺十二原穴对高、低血压大鼠模型血压影响的经穴效应规律研究.广州中医药大学博士论文,2008：68-69.
[48]Paxinos G, Watson C. The rat brain in stereotaxic coordinates. USA:Academic Press Inc, the sixth edition,2007:25-807.
[49]姚一康,杨伟敏,严国锋.对SHR大鼠病理特性测定研究.上海实验动物科学,2003,23(2)：75-82.
[50]宿中笑,肖达,睢久红,等.电针对自发性高血压大鼠血压和主动脉血管重构的影响.上海针灸杂志,2003,22(5)：20-24.
[51]杨华元,钟小红,刘堂义等.针刺仿真手法对高血压大鼠血压及心肌血管紧张素Ⅱ的影响.针刺研究,2008,33(3)：1186-1190.
[52]吴焕林,李晓庆,王侠.针刺太冲穴对65例肝阳上亢型高血压病患者的即时降压效应.中医杂志,2008,49(7)：622-624.
[53]尹艳茹.L-精氨酸对应激性大鼠血压的影响.中国临床药理学与治疗学,2002,7(6)：529-531.
[54]李漫松.侧脑室内注射BGN对应激性高血压大鼠心血管反应的影响.吉林大学学报(医学版),2003,29(4)：389-392.
[55]Kunz M, Scholl KE, Schu U, et al. GABAergic modulation of diffuse noxious inhibitory controls (DNIC):a test by use of lorazepam. Experimental Brain Research,2006,175(2):.363-371.
[56]王家有,唐纯志,赖新生,等.太冲穴中等强度针刺可降低高血压大鼠血压及血浆内皮素含量.针刺研究,2010,待刊.
[57]Tam K C, Yiu H H. The effect of acupuncture on essential hypertension. Am J Chin Med,1975,3(4):369-379.
[58]吴远华,朱广旗,林兴友,等.针刺曲池、太冲对高血压病患者血中ET和ACE的影响及疗效探讨.中国中西医结合杂志,2004,24(12)：1080-1083.
[59]Flachskampf, F A, Joachim Gallasch, Olaf Gefeller, et al. Randomized trial of acupuncture to lower blood pressure. Circulation,2007,115(24):3121-3129.
[60]Eric A. Macklin, Peter M.Wayne, Leslie A, et al. Stop Hypertension with the Acupuncture Research Program (SHARP):results of a randomized, controlled clinical trial[J]. Hypertension,2006.48(5):834-845.
[61]陈邦国,钱春艳,张静宁,等.针刺风池穴对高血压病降压疗效临床观察.上海针灸杂志,2006,25(3)：15-18.
[62]王侠,吴焕林,李晓庆.针刺太冲穴对动态血压的降压疗效观察.新中医,2007,39(11)：21-22.
[63]吴根诚,曹小定.针刺疗法走向世界的历史与影响因素即几点思考.中西医结合学报,2003,1(4)：247-251.
[64]Yin C, Seo B, Park HJ, et al. Acupuncture, a promising adjunctive therapy for essential hypertension:a double-blind, randomized, controlled trial. Neurol Res,2007,29 Suppl 1:98-104.
[65]李吉梅.针刺为主治疗顽固性高血压25例.河南中医,2005,25(1)：65-67.
[66]黄凡,姚国新,黄小丽,等.针刺治疗痰淤阻络型高血压病临床观察.中国针灸,2007,27(6)：403-406.
[67]王葳,李坤成,单保慈,等.针刺太冲穴的脑功能MRI研究.中华放射学杂志,2006,40(1)：29-35.
[68]Berman B M, Lao L, Langen berg P, et al. Effectiveness of acupuncture as adjunctive therapy in osteoarthritis of the knee:a randomized, controlled trial. Ann Intern Med,2004,141(12):901-910.
[69]Witt C,Brinkhaus B, Jena S, et al. Acupuncture in patients with ostearthritis of the knee:a randomised trial. Lancet,2005,366(9480): 136-143.
[70]艾林,戴建平,赵百孝,等.针刺镇痛机制的功能磁共振成像研究.中国医学影像技术,2004,20(8)：1197-1200.
[71]田小平,李瑛,梁繁荣,等.经穴与非经穴效应比较研究的现状与思考.中国针灸,2008,28(12)：907-909.
[72]胡卡明,王承平,谢慧君,等针刺穴位与非穴位激活不同脑区的效应观察.中国针灸,2006,26(3)：205-207.
[73]Maioli C, Falciati L, Marangon M, et al. Short-and long-term modulation of upper limb motor-evoked potentials induced by acupuncure. Eur J Neurosci,2006,23(1):1931-1938.
[74]李瑛,梁繁荣,郑晖,等.针刺经穴与非经非穴治疗偏头痛急性发作期的随机对照研究.世界传统医药国际科技大会论文集(针灸部分),2009：114-119
[75]Zhang W, LI JL, Hosaka M, et al. Cyclosporine A-induced hypertension involves synapsin in renal sensory nerve endings. Proc Natl Acad Sci USA,2000,97(17):9765-9770
[76]Koji Ito, Yoshitaka Hirooka, Nobuaki Hori, et al. Inhibition of Rho-Kinase in the nucleus tractus solitarius enhances glutamate sensitivity in rats. Hypertension online,2005, DOI:10.1161/01.
[77]张明亮,严志强,齐颖新,等.Rho蛋白解离抑制因子α在高血压大鼠胸主动脉的表达.解剖学报,2009,40(4)：625-629
[78]Wiedemann B, Franke W. Identification and localization of synaptophysin, an integral membrane glycoprotein of Mr 38,000 characteristic of presynapticvesicles.Cell,1985,41 (3):1017-1029
[79]Petra K, Heinrich B, Hubert R. Calcium-dependent serine phosphorylation of synaptophysin. Journal of Neur.2006,47(4):1302-1304.
[80]赵合庆,刘晶,余锋,等.经颅磁刺激对短暂性大脑中动脉闭塞大鼠突触素表达和超微结构改变的影响.中华脑血管并杂志(电子版),2008,2(1)：17-22
[81]沈丽,买鸿宴,南燕,等.自发性高血压大鼠中缝背核的超微结构.高血压杂志,1998,6(2)：209-213
[82]Evergren E, Benfenati F, Shupliakov O.The synapsin cycle:A view frm the synaptic endocytic zone. Neuroscience Reseach,2007,85(12):2648-2656.
[83]Lonart G, Simsek-Duran F. Deletion of synapsins I and II genes alters the size of vesicular pools and rabphilin phosphorylation. Bran Res,2006 1107:42-51
[84]Di Fraia R, Wilquet V, Ciardiello M A, et al.NADP+ dependent glutamate dehydrogenase in the antarctic psychrotolerant bacterium psychrobacter sp. Tadl. characterization, protein and DNA sequence and relationship to other glutamate dehydrogenases. Biochem,2000,267(1):121-131
[85]付海英,张纪周,杨宇丹,等.海洛因对大鼠谷氨酸脱氢酶基因表达的影响.中国药物依赖性杂志,2006,15(6)：440-444
[86]Cansein R O, Bonagamba L G, Machado B H. Bradycardic and hypotensive responses to microinjection of L-glutamate into the lateral aspect of the commissural NTS are blocked by an NMDA receptor antagonist. Bran Res,2000,852(1):68-75
[87]孟培燕,孙国杰,刘胜洪,等.电针预处理对血管性痴呆大鼠神经细胞谷氨酸-NMDA受体转导通路的影响.针刺研究,2008,33(2)：236-239
[88]崔毅军,汪石右.针刺对膀胱机能亢进大鼠蓝斑谷氨酸影响的动态研究.上海中医药大学博士学位论文,2005：2
[89]Rhian M. Touyz. Reactive Oxygen species, vascular oxidative stress,and redox signaling in hypertion. Hypertension,2004,44 (2):248-256
[90]Lie G, Wang W Z, Wang W, et al. Effects of angiotensin type 2 receptor overexpression in the rostral ventrolateral medulla on blood pressure and urine excretion in normal rats. Hypertension,2008,51 (2):521-527
[91]Leenen F H. Brain mechanisms contributing to sympathetic hyperactivity and heart failure. Circulation Res,2007,101 (3):221-223
[92]Ying W.NAD+/NADH and NADP+/NADPH in cellular functions and cell death:regulation and biological consequences. Antioxid Redox Signal, 2007,10 (2):179-206
[93]Infanger D W, Sharma R V, Davisson RL. NADPH oxidases of the brain:distribution, regulation and function. Axtioxid Redox Signal, 2006,8(2):1583-1596
[94]Freya Q. Schafer, Garry R, et al. Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radical Biology and Medicine,2001,30(11):1191-1212.
[95]周四桂,潘雪刁,唐富天,等.依那普利对肾性与自发性高血压大鼠左心室Enol及GSTM2表达的影响.中国药理学通报,2008,24(6)：736-740
[96]Iwai N, Tago N, Yasui N, et al. Analysis of 22 candidate genes for hypertension in the Japanese population. Hypertension,2004,22 (6):1119-1126.
[97]Custodis F, Eberl M, Kilter H, et al. Association of RhoGDIa with Racl GTPase mediates free radial production during myocardial hypertrophy. Cardiovasc Res,2006,71(2):342-351
[98]Pacaud P, Sauzeau V, Loirand G. Rho proteins and vascular diseases.Arch Mal Coeur Vaiss,2005,98(3):249-254
[99]Riha G M, Lin P H, Lumsden L B, et al. Roles of hemodynamic forces in vascular cell differentiation. Ann Biomech Eng,2005,33(6):772-779
[100]Pockley A G, Wu R, Lemne C, et al. Circulating heat shock protein 60 is associated with early cardiovascular disease. Hypertension,2000,36 (1): 303-307
[101]Amamoto K, Okamura T, Tamaki S, et al. Epidemiologic study of the association of low-Kmmitochondrial acetaldehyde dehydrogenase genotypes with blood pressure level and the prevalence of hypertension in a general population. Hypertension Res,2002,25(6):857-864
[102]Boggs J M. Myelin basic protein:a multifunctional protein. Cell Mol Life Sci,2006,63(1):1945-1961
[103]Maria Carolina Gongora, David G Harrison. Sad Heart from no SOD. Hypertension,2008,51:28-32
[104]Usmani S Z, Bona R, Li Z.17 AAG for HSP90 inhibition in cancer-from bench to bedside.Curr Mol Med,2009,9(5):654-664
[105]Moser C, Lang S A, Stoeltzing 0. Heat shock protein 90(Hsp90)as a molecular target for therapy of gastrointestinal cancer. Anticancer Res,2009,29(6):2031-2042
[106]Reikvam H, Ersvaer E, Bruserud 0. Heat shock protein 90 a potential target in the treatment of human acute myelogenous leukemia. Curr Cancer Drug Targets,2009,9(6):761-776
[107]Tsutsumi S, Beebe K, Neckers L. Impact of heat-shock protein 90 on cancer metastasis. Future Oncol,2009,5 (5):679-688
[108]Moleda L, Jurzik L, Froh M, et al.Role of HSP-90 for increased nNOS-mediated vasodilation in mesenteric arteries in portal hypertension. World J Gastroenterol,2010,16(15):1837-1844
[109]Moiseeva 0 M, Novoselov S S, Liasnikova E A, et al.Heat shock protein 70 as a marker of lesions to target organs in hypertension.Vestn Ross Akad Med Nauk,2004,1(1):6-9
[110]Wright B H, Corton J M, E1-Nahas A M, et al. Elevated levels of circulating heat shock protein 70 in peripheral and renal vascular disease. Heart Vessels,2000,15(1):18-22
[111]Pockley A G, De Faire U, Kiessling R, et al. Circulating heat shock protein and heat shock protein antibody levels in established hypertension. Hypertension,2002,20 (1):1815-1829
[112]Moon K H, Kim B J, Song B J.Inhibition of mitochondrial aldehyde dehydro-genase by nitric oxide-mediated S-nitrosylation. Febs Lett,2005,579(27):6115-6120
[113]Daiber A, Wenzel P, Oelze M, et al. Mitochondrial aldehyde dehydrogenase (ALDH-2)-marker of and marker for nitrate tolerance in response to nitroglycer in treatment. Chem Biol Int,2009,178(1-3):40-47
[114]Chen Z Q, Stamler J S. Bioactivaction of nitroglycerin by the mitochondrial aldehyde dehydrogenase. Trends Cardiovasc Med,2006,16(8):259-265
[115]Sun A J, Wang K Q, Ge J B, et al. Proteomic analysis of myocardial mitochondria in rats with heart failure. Cardiovasc Drugs Ther,2004,18(1):21-29
[116]Hui P, Nakayama T, Morita A, et al. Common single nucleotide polymorphisms in Japanese patients with essential hypertension:aldehyde dehydrogenase 2 gene as a risk factor independent of alcohol consumption. Hypertension Res,2007,30(7):585-592
[117]韩济生.神经科学原理[M].第2版,北京：北京医科大学出版社,1999：1050-1059
[118]乔松,冯加纯,杨晶,等.高血压大鼠脑内小血管改变及其继发损伤的研究.中风与神经疾病杂志,2006,23(3)：290-294
[119]宿宝贵,潘三强,,王家有,等.慢性肾性高血压大鼠脑组织磁共振成像观察和超微结构的研究.解剖学报,2005,35(6)：636-639
[120]Constantinescu C S,Goodman D B,Hilliard B, ea al.Murine macrophages stimulated with central and peripheral nervous system myelin or purified myelin proteins release inflammatory products. Neurosci Lett.2000,287 (3):171-174
[121]刘红岩,杨贵贞.髓磷脂碱性蛋白促进单个核细胞对内皮细胞的粘附效应.中国免疫学杂志,2000,16(8)：428-431
[122]Hidefumi Waki,Beihui Liu, Massao Miyake,et al. Junctional adhesion molecule-1 is upregulated in spontaneously hypertensive rats. Hypertension, 2007,49:1321-1327
[1]严季澜.建国40年来高血压病的针灸推拿治疗概况[J].天津中医,1990,(6)：37.
[2]佘延芬,于文,夏茜,等.近10年针灸治疗高血压病的研究进展[J].河北中医药学报,2000,15(3)：36.
[3]刘锦,乔嘉斌.针刺治疗高血压的研究进展[J].吉林中医药,2008,28(2)：152.
[4]Tam K C, Yiu H H. The effect of acupuncture on essential hypertension[J]. Am J Chin Med,1975,3(4):369.
[5]陈邦国,钱春艳,张静宁,等.针刺风池穴对高血压病降压疗效临床观察[J].上海针灸杂志,2006,25(3)：15.
[6]卫彦,孙忠人,寇吉友,等.针刺人迎穴治疗高血压病120例临床观察[J].针灸临床杂志,2006,22(2)：4.
[7]万文俊,马朝阳,熊修安,等.电针曲池穴治疗原发性高血压病疗效观察及机制探讨.中国针灸,2009,29(5)：349.
[8]Yin C, Seo B, Park HJ,et al. Acupuncture, a promising adjunctive therapy for essential hypertension:a double-blind, randomized, controlled trial[J]. Neurol Res,2007,29 Suppl 1:98.
[9]李吉梅.针刺为主治疗顽固性高血压25例[J].河南中医,2005,25(1)：65.
[10]黄凡,姚国新,黄小丽,等.针刺治疗痰淤阻络型高血压病临床观察.中国针灸,2007,27(6)：403.
[11]吴远华,朱广旗,林兴友,等.针刺曲池、太冲对高血压病患者血中ET和ACE的影响及疗效探讨.中国中西医结合杂志,2004,24(12)：1080.
[12]Flachskampf, F A, Joachim Gallasch, Olaf Gefeller, et al. Randomized trial of acupuncture to lower blood pressure[J]. Circulation,2007,115(24):3121.
[13]郭玉红.针刺治疗高血压病胰岛素抵抗的临床观察[J].中医药学报,2007,35(6)：51.
[14]Sugioka, K, Mao W, Woods J, et al. An unsuccessful attempt to treat hypertension with acupuncture[J]. Am J Chin Med (Gard City N Y),1977,5(1):39.
[15]Eric A. Macklin, Peter M.Wayne, Leslie A, et al. Stop Hypertension with the Acupuncture Research Program (SHARP):results of a randomized, controlled clinical trial[J]. Hypertension,2006.48(5):838.
[16]Lee H, Kim S Y, Park J, et al. Acupuncture for lowering blood pressure: systematic review and meta-analysis[J]. Am J Hypertens,2009,22:122.
[17]吴焕林.邓铁涛教授治疗高血压病临床经验辑要[J].河南中医,2005,25(5)：16.
[18]王侠,邹旭,李琼,等.针刺太冲穴治疗高血压的临床研究[J].实用医学杂志,2003,19(5)：565.
[19]朱广旗,吴远华,吴邦启,等.针刺曲池和太冲对高血压病不同证型的疗效[J].浙江中西医结合杂志,2006,16(1)：4.
[20]穆晓红,李巧霞.针刺太冲和足三里治疗高血压病临床观察[J].上海针灸杂志,2009,28(6)：328.
[21]张为.针刺悬钟穴治疗高血压病40例[J].河北中医,2006,28(2)：154.
[22]曹玲,侯广云,范文双.针刺颈部夹脊穴治疗颈源性高血压的疗效分析[J].针刺研究,2007,32(3)：195.
[23]吴焕林,李晓庆,王侠.针刺太冲穴对65例肝阳上亢型高血压病患者的即时降压效应[J].中医杂志,2008,49(7)：622.
[24]冯国湘,吴清明.针刺开“四关”加百会穴温针灸治疗原发性高血压的临床研究[J].中国针灸,2003,23(4)：193.
[25]刘建永,杨佃会,白田雨.针灸治疗原发性高血压临床用穴频次分析[J].山东中医药大学学报,2009,33(3)：215.
[26]杨路,赖新生.针刺十二原穴对高、低血压大鼠模型血压影响的经穴效应规律研究[D].广州中医药大学博士论文,2008：68
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