电针镇痛的累积效应与下丘脑、海马胆碱能神经元关系的研究
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摘要
研究背景
针刺治疗疾病具有累积效应现在已是众多研究者所公认的事实。但是目前关于针刺累积效应的具体机理尚不明确,未见到系统的研究报道。已有的动物实验结果表明:反复针刺所产生的累积效应与神经系统的特别是中枢神经系统对针刺信息的储存、编码等活动密切相关;下丘脑、海马等边缘系统、中枢胆碱能系统参与针刺镇痛的过程。我们实验室前期的工作证明大鼠下丘脑、海马囊泡乙酰转移酶(VAChT)、胆碱乙酰转移酶(ChAT)等参与针刺镇痛的累积效应;去卵巢(OVX)引起的记忆功损伤减弱了针刺的效应。为了从不同的环节进一步确认下丘脑、海马胆碱能神经元在针刺累积性镇痛效应中的作用,本研究在结扎坐骨神经造成压迫性损伤(chronic constrictive injury,CCI)/及OVX大鼠记忆损害的动物模型上,从乙酰胆碱降解酶胆碱酯酶(AChE)和细胞内蛋白激酶PKA的活性的改变、以及毒蕈碱M_1受体m RNA表达的变化,探讨针刺累积效应的机制。
研究方法
Wistar雌性大鼠103只,随机分为正常对照组(n=15),慢性压迫损伤(CCI)组(n=15),CCI+EA2天(D)组(n=15),CCI+EA2周(W)组(n=15),去卵巢(OVX)+CCI(n=13),OVX+CCI+EA2D(n=15),OVX+CCI+EA2W(n=15)组。Morris水迷宫测试检查OVX+D-半乳糖皮下注射动物学习记忆能力的变化。结扎坐骨神经造成CCI慢性痛模型,电针双侧“足三里”-“阳陵泉”穴(2/15Hz,1-2mA,1次/D)。用辐射热照射大鼠双侧足底引起的缩腿反应潜伏期(PWL)的差值作为痛敏分数(HS)判断动物的痛反应。在麻醉状态下,一部分(每组8-10例)动物常规灌流取脑,冰冻切片,用酶组织化学方法来检测下丘脑弓状核(ARC)区、视上核(SON)区和室旁核(PVN)区,及海马CA1区AChE的表达;用免疫组化法检测其PKA活性。另一部分动物(每组5例)取脑组织匀浆后,用RT-PCR的方法来检测M_1受体m RNA的表达的变化。
结果
1)电针对CCI大鼠痛阈变化的影响
CCI术18天后,与正常组(0.22±0.68)比较,各组HS的绝对值都明显升高(P＜0.05)。在单纯CCI动物上,与CCI对照组(-4.22±0.33)比,CCI术后第18天CCI+EA2D(-2.76±0.32)、CCI+EA2W组(-0.42±0.68)HS的绝对值显著减小(P＜0.05);CCI+EA2W组(-0.42±0.68)HS绝对值明显低于CCI+EA2D组(-2.76±0.32,P＜0.05)。在OVX动物上,OVX+CCI+EA2W组(-2.37±0.29)和CCI+EA2D组(-3.11±0.32)HS绝对值明显低于OVX+CCI组(-4.66±0.45,P＜0.05);OVX+CCI+EA2W组HS绝对值(-2.37±0.29)明显低于CCI+EA2D组(-3.11±0.32,P＜0.05)。提示电针可以改善CCI动物的痛敏状态,并具有累加性。CCI+EA2W组HS的绝对值(-0.42±0.68)显著低于OVX+CCI+EA2W组(-2.37±0.29,P＜0.05),说明记忆损伤动物的电针累积效应明显降低。
2)电针对CCI大鼠下丘脑三核团、海马组织胆碱能神经纤维AChE的活性变化的影响:
坐骨神经结扎后,CCI组ARC(144.43±4.68)、SON(135.14±6.95)AChE的表达显著少于正常对照组ARC(126.27±4.35)、SON(122.2±7.5)(P＜0.05,灰度值越小,表达量越大)。电针2天后,CCI+EA2D组AChE的表达在ARC(133.32±8.03)、SON(111.24±6.01)显著增多(P＜0.05)。CCI+EA2W组下丘脑AChE的表达PVN(100.11±12.71)、ARC(117.25±4.12)和SON区(95.27±7.23)显著多于CCI对照组和CCI+EA2D组(P＜0.05)。复合模型组中的表达趋势与单纯CCI模型组相同,电针后AChE的表达累积性上调。OVX+CCI+EA2W组AChE的表达ARC(133.23±11.48)、和SON区(113.92±7.11)均显著少于CCI+EA2W组同一脑区(P＜0.05)。海马CA1区各组AChE的表达与下丘脑区各组的表达趋势一致。结果说明:CCI后,下丘脑和海马组织AChE的活性有所抑制,电针可使AChE的活性增强,并有累积性上调;去除卵巢,动物记忆能力受到损害后,仍有累积效应存在,但明显减弱。
3)电针对CCI大鼠下丘脑、海马组织毒蕈碱M_1受体mRNA表达的影响
CCI后,下丘脑组织M_1受体mRNA的表达量(0.39±0.25)与正常对照组(0.83±0.40)比较显著性降低(P＜0.05);CCI+EA2W组M_1受体mRNA表达量(0.88±0.16)显著高于CCI对照组(0.39±0.25)和CCI+EA2D组(0.59±0.21,P＜0.05)。记忆损伤动物中,OVX+CCI组下丘脑M_1受体mRNA的表达量(0.29±0.11)明显少于正常对照组(0.83±0.40,P＜0.05);OVX+CCI+EA2W组M_1受体mRNA的表达量(0.76±0.35)显著高于OVX+CCI(0.29±0.11)和OVX+CCI+EA2D组(0.41±0.22,P＜0.05)。这说明电针可以累积性上调M_1受体mRNA的表达。海马区M_1受体mRNA的表达趋势与下丘脑区的表达变化趋势类似:OVX+CCI+EA2W组(1.00±0.20)M_1受体mRNA的表达显著少于CCI+EA2W(1.37±0.37),提示电针上调M_1受体mRNA表达的累积作用记忆损伤后减弱。
4)电针对CCI大鼠下丘脑三核团、海马CA1区神经元PKA的表达变化的影响
CCI+EA2W组下丘脑PVN(99.28±19.33)、ARC(100.18±14.52)和SON(103.26±17.12)区PKA的表达均明显高于正常对照组PVN(133.54±6.31),ARC(127.84±18.91),SON(133.53±21.34)(P＜0.05;灰度值越小,表达量越大),也显著高于CCI+EA2D组PVN(124.35±6.3),ARC(121.77±14.66),SON(118.61±9.64,P＜0.05)。OVX+CCI+EA2W组PVN(107.48±12.39)、ARC(117.11±19.87)和SON(129.12±16.96)区PKA表达亦均明显高于OVX+CCI组PVN(145.96±8.02),ARC(134.24±18.43),SON(148.42±10.7(P＜0.05),说明EA2W可使PKA的表达显著上调,且有累积性作用。OVX+CCI+EA2W组ARC和SON区PKA的表达均显著低于CCI+EA2W组(P＜0.05),提示EA上调PKA表达的累积效应在记忆功能减退的动物上明显减弱。海马PKA的表达与SON等下丘脑核团PKA的表达趋势类似。
结论
1)电针“足三里”-“阳陵泉”可明显减轻CCI后出现的痛反应,且有累积效应;记忆损伤后减弱电针的效应;
2)电针可显著减轻CCI引起的下丘脑、海马AChE、毒蕈碱M_1受体mRNA表达的抑制作用,累积性上调AChE、PKA及M_1受体mRNA的表达;记忆损伤后电针的上述效应降低。
因此,电针“足三里”-“阳陵泉”减轻慢性痛的累积效应与其上调下丘脑和海马组织中AChE、M_1受体mRNA、PKA的表达作用、与动物神经记忆功能有密切联系。本研究结果为反复针刺治疗产生的累积效应提供了系统而详实的实验依据。
Background
It is a generally acknowledged that repeated acupuncture stimulation of acupoints has an accumulative effect in the treatment of diseases.But its underlying mechanism remains unknown,and no any systematic studies have been reported up to now.Experimental studies have indicated that the cumulative effect of repeated acupuncture has a close relationship with the nervous system,especially with the processing,storage,and coding of the central nervous system.The limbic system including hypothalamus and hippocampus,and the central cholinergic system have been shown to participate in the process of the accumulative analgesic effect of electroacupuncture(EA).Our research results have revealed that hypothalamic and hippocampal neuronal vesicles acetyltransferase (VAChT),choline acetyltransferase(CHAT) play a certain role in EA-induced accumulative analgesic effect in chronic constrictive injury (CCI) rats;and the memory impairment caused by ovariectomized(OVX) may weaken the effects of acupuncture.In order to confirm the relationship between the accumulative analgesic effect of EA and the cholinergic neurons in the hypothalamus and hippocampus from different links,the present study was designed to further explore the mechanism underlying EA-induced accumulative analgesic effect from the activity of acetylcholinesterase(ACHE,a catabolic enzyme of acetylcholine) and intracellular protein kinase A(PKA),and changes of muscarinic M_1 receptor mRNA expression and neuron-memory in rats with chronic neuropathy pain and or OVX-induced memory impairment.
Materials and Methods
A hundred and three female Wistar rats(230-250g) were randomized into normal control,CCI,CCI+EA2D,CCI+EA2W,OVX+CCI,OVX+CCI+EA2D and OVX+CCI+EA2W groups,with 15 cases in each except OVX+CCI group(n=13). Neuro-memory impairment was induced by OVX plus subcutaneous injection of D-galactose.CCI pain model was established by ligature of the right sciatic nerve with surgical suture.EA(2/15Hz,1-2 mA,30 min) was daily applied to bilateral "Zusanli"(ST36) and "Yanglingquan"(GB34) for 2 days(2D) and 2 weeks(2W) respectively.Hyperalgesic scores(HS) were detected with radiation-heat irradiation.After deeply anesthetized(25% urethane and 1.5%cholorase,0.5ml/100g),the rats(8-10/group) were perfused transcardially and the brain tissue was dissected out to be sectioned at 60μm on a freezing microtome.AChE activity and PKA activity of hypothalamic arcuate nucleus(ARC),supraoptic nucleus(SON) and paraventricular nucleus(PVN) and Hippocampal CA1 of different groups were measured respectively with the enzyme histochemistry and immunohistochemical methods.Other 35 anesthetized rats(5/group) were executed on the ice-plate and their brain tissue was taken out.Muscarinic M_1 receptor mRNA expression was detected by reversed transcription-polymerase chain reaction(RT-PCR) after homogenate.
Results:
1) Effect of EA on pain threshold in CCI rats
After CCI,the absolute values of HS of all groups increased significantly in comparison with normal control group(0.22±0.68, P＜0.05).In simple CCI rats,compared with CCI group(-4.22±0.33),the absolute values of HS of both CCI+EA2D(-2.76±0.32) and CCI+EA2W (-0.42±0.68) groups decreased markedly(P＜0.05),and that of CCI+EA2W group(-0.42±0.68) was evidently lower than that of CCI+EA2D group (-2.76±0.32,P＜0.05) on the 18th day after CCI(P＞0.05).In memory impairment rats,the absolute values of the HS of both OVX+CCI+EA2W (-2.37±0.29)and CCI+EA2D(-3.11±0.32)groups were significantly lower than that of OVX+CCI group(-4.66±0.45,P＜0.05).Compared with OVX+CCI+EA2D(-3.11±0.32) group,the absolute values of HS of OVX+CCI+EA2W(-2.37±0.29) group decreased markedly(P＜0.05),suggesting that electroacupuncture treatment can improve the animal's pain sensitivity after CCI,and has an accumulative effect.The absolute value of HS of CCI+EA2W(-0.42±0.68) was apparently lower than that of OVX+CCI+EA2W group(-2.37±0.29,P＜0.05),indicating that the memory impairment weakens the above mentioned effects of EA.
2) Effect of EA on AChE activity of cholinergic nerve fibers in three nuclei of hypothalamus and hippocampus in CCI model rats
After CCI,AChE expression of ARC(144.43±4.68) and SON(135.14±6.95) in CCI group were significantly lower than ARC(126.27±4.35),SON (122.2±7.5) in normal control group(P＜0.05,the bigger the integral grey value,the less the expression of ACHE).In comparison with normal control group,AChE expression in the ARC(133.32±8.03) and SON(111.24±6.01) in CCI+EA2D group increased significantly(P＜0.05).Compared with CCI control group and CCI+EA2D group,AChE expression in PVN(100.11±12.71),ARC(117.25±4.12) and SON(95.27±7.23) in CCI+EA2W group increased significantly(P＜0.05).AChE expression in ARC,SON and PVN in OVX+CCI,OVX+CCI+EA2D and OVX+CCI+EA2W groups presented the similar tendency in comparison with CCI,CCI+EA2D and CCI+EA2W groups after the EA treatment,and EA-induced upregulation of AChE expression has a cumulative efficacy.AChE expression of OVX+CEI+EA2W group in ARC(133.23±11.48) and SON(113.92±7.11) was significantly lower than that of CCI + EA2W group in the same brain regions(P＜0.05).AChE expression in hippocampal CA1 area showed the similar tendency.These results showed that after CCI,the activity of AChE was down-regulated in hypothalamus and hippocampus,EA of ST36-GB34 can increase the activity of ACHE,and induce a cumulative increase;OVX-induced memory impairment(estrin level-decrease) attenuates EA-induced effects,in spite of still existing in OVX rats.
3) Effect of EA on muscarinic M_1 receptor mRNA expression in hypothalamus and hippocampus in CCI rats
After CCI,M_1 receptor mRNA expression(0.39±0.25) in hypothalamus was considerably lower than that of normal control group(0.83±0.40, P＜0.05).Compared with CCI control group(0.39±0.25) and CCI+EA2D group (0.59±0.21),M_1 receptor mRNA expression of CCI+EA2W group(0.88±0.16) was increased significantly(P＜0.05).In OVX+CCI model,M_1 receptor mRNA expression of OVX+CCI control group(0.29±0.11) was markedly lower than that of normal control group(0.83±0.40,P＜0.05).Compared with OVX+CCI (0.29±0.11) and OVX+CCI+EA2D group(0.41±0.22),the expression of M_1 receptor mRNA expression(0.76±0.35) in OVX+CCI+EA2W group was markedly increased(P＜0.05),suggesting a cumulative increase of M_1 receptor mRNA expression after EA treatment.The expression of M_1 receptor mRNA in hippocampus tissue had a similar trend in comparison with hypothalamus tissue.The expression of M_1 receptor mRNA in OVX+CCI+EA2W group (1.00±0.20) was lower than that of CCI+EA2W group (1.37±0.37),suggesting an attenuation of EA-induced cumulative effect in up-regulating M_1 receptor mRNA expression after memory impairment in OVX rats.
4) Effect of EA on PKA activity in hypothalamus and hippocampus in CCI rats
Results of immunohistochemistry showed that the expressions of PKA of PVN(99.28±19.33,integral grey value),ARC(100.18±14.52) and SON (103.26±17.12) of hypothalamus in CCI+EA2W group were significantly higher than those in PVN(133.54±6.31),ARC(127.84±18.91) and SON (133.53±21.34) in normal control group;and PVN(124.35±6.3),ARC (121.77±14.66),SON(118.61±9.64) in CCI+2D group(P＜0.05).PKA expressions of PVN(107.48±12.39),ARC(117.11±19.87) and SON (129.12±16.96) in OVX+CCI+EA2W group were also markedly higher than PVN (145.96±8.02),ARC(134.24±18.43) and SON(148.42±10.7,P＜0.05) in OVX+CCI group,displaying upregulation and cumulative increase of PKA expression after EA2W.PKA expression of PKA of ARC and SON in OVX+CCI+EA2W group was markedly lower than that in CCI+EA2W group(P＜0.05),suggesting an apparent attenuation of EA-induced cumulative upregulation effect of PKA expression in OVX rats.The changing tendency of PKA expression in hippocampal CA1 region was similar to that of hypothalamic SON after repeated EA of ST36-GB34 in CCI and OVX+CCI rats.
Conclusion
1) EA of "Zusanli"(ST36)-"Yanglingquan"(GB34) is able to significantly alleviate pain sensitivity caused by CCI,and has a cumulative effect,which is weakened obviously after neuron-memory injury.
2) EA can significantly reduce CCI-induced inhibition of the expression of ACHE,M_1 receptor mRNA in hypothalamus and hippocampus,and has an accumulative upregulation effect on ACHE,PKA and M_1 receptor mRNA expression,which are closely associated with the animals' neuron-memory ability.
Therefore,there is a close relationship between the accumulative analgesic effect of EA and its resultant increase of the expression of ACHE,M_1 receptor mRNA and PKA in hypothalamus and hippocampus,and the function of neuron-memory is involved in the cumulative effect of repeated EA treatment.Results of the present study provide a set of detailed experimental evidence for repeated acupuncture-induced cumulative effect in the treatment of chronic pain.
针刺治疗疾病具有累积效应现在已是众多研究者所公认的事实。但是目前关于针刺累积效应的具体机理尚不明确,未见到系统的研究报道。已有的动物实验结果表明:反复针刺所产生的累积效应与神经系统的特别是中枢神经系统对针刺信息的储存、编码等活动密切相关;下丘脑、海马等边缘系统、中枢胆碱能系统参与针刺镇痛的过程。我们实验室前期的工作证明大鼠下丘脑、海马囊泡乙酰转移酶(VAChT)、胆碱乙酰转移酶(ChAT)等参与针刺镇痛的累积效应;去卵巢(OVX)引起的记忆功损伤减弱了针刺的效应。为了从不同的环节进一步确认下丘脑、海马胆碱能神经元在针刺累积性镇痛效应中的作用,本研究在结扎坐骨神经造成压迫性损伤(chronic constrictive injury,CCI)/及OVX大鼠记忆损害的动物模型上,从乙酰胆碱降解酶胆碱酯酶(AChE)和细胞内蛋白激酶PKA的活性的改变、以及毒蕈碱M_1受体m RNA表达的变化,探讨针刺累积效应的机制。
研究方法
Wistar雌性大鼠103只,随机分为正常对照组(n=15),慢性压迫损伤(CCI)组(n=15),CCI+EA2天(D)组(n=15),CCI+EA2周(W)组(n=15),去卵巢(OVX)+CCI(n=13),OVX+CCI+EA2D(n=15),OVX+CCI+EA2W(n=15)组。Morris水迷宫测试检查OVX+D-半乳糖皮下注射动物学习记忆能力的变化。结扎坐骨神经造成CCI慢性痛模型,电针双侧“足三里”-“阳陵泉”穴(2/15Hz,1-2mA,1次/D)。用辐射热照射大鼠双侧足底引起的缩腿反应潜伏期(PWL)的差值作为痛敏分数(HS)判断动物的痛反应。在麻醉状态下,一部分(每组8-10例)动物常规灌流取脑,冰冻切片,用酶组织化学方法来检测下丘脑弓状核(ARC)区、视上核(SON)区和室旁核(PVN)区,及海马CA1区AChE的表达;用免疫组化法检测其PKA活性。另一部分动物(每组5例)取脑组织匀浆后,用RT-PCR的方法来检测M_1受体m RNA的表达的变化。
结果
1)电针对CCI大鼠痛阈变化的影响
CCI术18天后,与正常组(0.22±0.68)比较,各组HS的绝对值都明显升高(P＜0.05)。在单纯CCI动物上,与CCI对照组(-4.22±0.33)比,CCI术后第18天CCI+EA2D(-2.76±0.32)、CCI+EA2W组(-0.42±0.68)HS的绝对值显著减小(P＜0.05);CCI+EA2W组(-0.42±0.68)HS绝对值明显低于CCI+EA2D组(-2.76±0.32,P＜0.05)。在OVX动物上,OVX+CCI+EA2W组(-2.37±0.29)和CCI+EA2D组(-3.11±0.32)HS绝对值明显低于OVX+CCI组(-4.66±0.45,P＜0.05);OVX+CCI+EA2W组HS绝对值(-2.37±0.29)明显低于CCI+EA2D组(-3.11±0.32,P＜0.05)。提示电针可以改善CCI动物的痛敏状态,并具有累加性。CCI+EA2W组HS的绝对值(-0.42±0.68)显著低于OVX+CCI+EA2W组(-2.37±0.29,P＜0.05),说明记忆损伤动物的电针累积效应明显降低。
2)电针对CCI大鼠下丘脑三核团、海马组织胆碱能神经纤维AChE的活性变化的影响:
坐骨神经结扎后,CCI组ARC(144.43±4.68)、SON(135.14±6.95)AChE的表达显著少于正常对照组ARC(126.27±4.35)、SON(122.2±7.5)(P＜0.05,灰度值越小,表达量越大)。电针2天后,CCI+EA2D组AChE的表达在ARC(133.32±8.03)、SON(111.24±6.01)显著增多(P＜0.05)。CCI+EA2W组下丘脑AChE的表达PVN(100.11±12.71)、ARC(117.25±4.12)和SON区(95.27±7.23)显著多于CCI对照组和CCI+EA2D组(P＜0.05)。复合模型组中的表达趋势与单纯CCI模型组相同,电针后AChE的表达累积性上调。OVX+CCI+EA2W组AChE的表达ARC(133.23±11.48)、和SON区(113.92±7.11)均显著少于CCI+EA2W组同一脑区(P＜0.05)。海马CA1区各组AChE的表达与下丘脑区各组的表达趋势一致。结果说明:CCI后,下丘脑和海马组织AChE的活性有所抑制,电针可使AChE的活性增强,并有累积性上调;去除卵巢,动物记忆能力受到损害后,仍有累积效应存在,但明显减弱。
3)电针对CCI大鼠下丘脑、海马组织毒蕈碱M_1受体mRNA表达的影响
CCI后,下丘脑组织M_1受体mRNA的表达量(0.39±0.25)与正常对照组(0.83±0.40)比较显著性降低(P＜0.05);CCI+EA2W组M_1受体mRNA表达量(0.88±0.16)显著高于CCI对照组(0.39±0.25)和CCI+EA2D组(0.59±0.21,P＜0.05)。记忆损伤动物中,OVX+CCI组下丘脑M_1受体mRNA的表达量(0.29±0.11)明显少于正常对照组(0.83±0.40,P＜0.05);OVX+CCI+EA2W组M_1受体mRNA的表达量(0.76±0.35)显著高于OVX+CCI(0.29±0.11)和OVX+CCI+EA2D组(0.41±0.22,P＜0.05)。这说明电针可以累积性上调M_1受体mRNA的表达。海马区M_1受体mRNA的表达趋势与下丘脑区的表达变化趋势类似:OVX+CCI+EA2W组(1.00±0.20)M_1受体mRNA的表达显著少于CCI+EA2W(1.37±0.37),提示电针上调M_1受体mRNA表达的累积作用记忆损伤后减弱。
4)电针对CCI大鼠下丘脑三核团、海马CA1区神经元PKA的表达变化的影响
CCI+EA2W组下丘脑PVN(99.28±19.33)、ARC(100.18±14.52)和SON(103.26±17.12)区PKA的表达均明显高于正常对照组PVN(133.54±6.31),ARC(127.84±18.91),SON(133.53±21.34)(P＜0.05;灰度值越小,表达量越大),也显著高于CCI+EA2D组PVN(124.35±6.3),ARC(121.77±14.66),SON(118.61±9.64,P＜0.05)。OVX+CCI+EA2W组PVN(107.48±12.39)、ARC(117.11±19.87)和SON(129.12±16.96)区PKA表达亦均明显高于OVX+CCI组PVN(145.96±8.02),ARC(134.24±18.43),SON(148.42±10.7(P＜0.05),说明EA2W可使PKA的表达显著上调,且有累积性作用。OVX+CCI+EA2W组ARC和SON区PKA的表达均显著低于CCI+EA2W组(P＜0.05),提示EA上调PKA表达的累积效应在记忆功能减退的动物上明显减弱。海马PKA的表达与SON等下丘脑核团PKA的表达趋势类似。
结论
1)电针“足三里”-“阳陵泉”可明显减轻CCI后出现的痛反应,且有累积效应;记忆损伤后减弱电针的效应;
2)电针可显著减轻CCI引起的下丘脑、海马AChE、毒蕈碱M_1受体mRNA表达的抑制作用,累积性上调AChE、PKA及M_1受体mRNA的表达;记忆损伤后电针的上述效应降低。
因此,电针“足三里”-“阳陵泉”减轻慢性痛的累积效应与其上调下丘脑和海马组织中AChE、M_1受体mRNA、PKA的表达作用、与动物神经记忆功能有密切联系。本研究结果为反复针刺治疗产生的累积效应提供了系统而详实的实验依据。
Background
It is a generally acknowledged that repeated acupuncture stimulation of acupoints has an accumulative effect in the treatment of diseases.But its underlying mechanism remains unknown,and no any systematic studies have been reported up to now.Experimental studies have indicated that the cumulative effect of repeated acupuncture has a close relationship with the nervous system,especially with the processing,storage,and coding of the central nervous system.The limbic system including hypothalamus and hippocampus,and the central cholinergic system have been shown to participate in the process of the accumulative analgesic effect of electroacupuncture(EA).Our research results have revealed that hypothalamic and hippocampal neuronal vesicles acetyltransferase (VAChT),choline acetyltransferase(CHAT) play a certain role in EA-induced accumulative analgesic effect in chronic constrictive injury (CCI) rats;and the memory impairment caused by ovariectomized(OVX) may weaken the effects of acupuncture.In order to confirm the relationship between the accumulative analgesic effect of EA and the cholinergic neurons in the hypothalamus and hippocampus from different links,the present study was designed to further explore the mechanism underlying EA-induced accumulative analgesic effect from the activity of acetylcholinesterase(ACHE,a catabolic enzyme of acetylcholine) and intracellular protein kinase A(PKA),and changes of muscarinic M_1 receptor mRNA expression and neuron-memory in rats with chronic neuropathy pain and or OVX-induced memory impairment.
Materials and Methods
A hundred and three female Wistar rats(230-250g) were randomized into normal control,CCI,CCI+EA2D,CCI+EA2W,OVX+CCI,OVX+CCI+EA2D and OVX+CCI+EA2W groups,with 15 cases in each except OVX+CCI group(n=13). Neuro-memory impairment was induced by OVX plus subcutaneous injection of D-galactose.CCI pain model was established by ligature of the right sciatic nerve with surgical suture.EA(2/15Hz,1-2 mA,30 min) was daily applied to bilateral "Zusanli"(ST36) and "Yanglingquan"(GB34) for 2 days(2D) and 2 weeks(2W) respectively.Hyperalgesic scores(HS) were detected with radiation-heat irradiation.After deeply anesthetized(25% urethane and 1.5%cholorase,0.5ml/100g),the rats(8-10/group) were perfused transcardially and the brain tissue was dissected out to be sectioned at 60μm on a freezing microtome.AChE activity and PKA activity of hypothalamic arcuate nucleus(ARC),supraoptic nucleus(SON) and paraventricular nucleus(PVN) and Hippocampal CA1 of different groups were measured respectively with the enzyme histochemistry and immunohistochemical methods.Other 35 anesthetized rats(5/group) were executed on the ice-plate and their brain tissue was taken out.Muscarinic M_1 receptor mRNA expression was detected by reversed transcription-polymerase chain reaction(RT-PCR) after homogenate.
Results:
1) Effect of EA on pain threshold in CCI rats
After CCI,the absolute values of HS of all groups increased significantly in comparison with normal control group(0.22±0.68, P＜0.05).In simple CCI rats,compared with CCI group(-4.22±0.33),the absolute values of HS of both CCI+EA2D(-2.76±0.32) and CCI+EA2W (-0.42±0.68) groups decreased markedly(P＜0.05),and that of CCI+EA2W group(-0.42±0.68) was evidently lower than that of CCI+EA2D group (-2.76±0.32,P＜0.05) on the 18th day after CCI(P＞0.05).In memory impairment rats,the absolute values of the HS of both OVX+CCI+EA2W (-2.37±0.29)and CCI+EA2D(-3.11±0.32)groups were significantly lower than that of OVX+CCI group(-4.66±0.45,P＜0.05).Compared with OVX+CCI+EA2D(-3.11±0.32) group,the absolute values of HS of OVX+CCI+EA2W(-2.37±0.29) group decreased markedly(P＜0.05),suggesting that electroacupuncture treatment can improve the animal's pain sensitivity after CCI,and has an accumulative effect.The absolute value of HS of CCI+EA2W(-0.42±0.68) was apparently lower than that of OVX+CCI+EA2W group(-2.37±0.29,P＜0.05),indicating that the memory impairment weakens the above mentioned effects of EA.
2) Effect of EA on AChE activity of cholinergic nerve fibers in three nuclei of hypothalamus and hippocampus in CCI model rats
After CCI,AChE expression of ARC(144.43±4.68) and SON(135.14±6.95) in CCI group were significantly lower than ARC(126.27±4.35),SON (122.2±7.5) in normal control group(P＜0.05,the bigger the integral grey value,the less the expression of ACHE).In comparison with normal control group,AChE expression in the ARC(133.32±8.03) and SON(111.24±6.01) in CCI+EA2D group increased significantly(P＜0.05).Compared with CCI control group and CCI+EA2D group,AChE expression in PVN(100.11±12.71),ARC(117.25±4.12) and SON(95.27±7.23) in CCI+EA2W group increased significantly(P＜0.05).AChE expression in ARC,SON and PVN in OVX+CCI,OVX+CCI+EA2D and OVX+CCI+EA2W groups presented the similar tendency in comparison with CCI,CCI+EA2D and CCI+EA2W groups after the EA treatment,and EA-induced upregulation of AChE expression has a cumulative efficacy.AChE expression of OVX+CEI+EA2W group in ARC(133.23±11.48) and SON(113.92±7.11) was significantly lower than that of CCI + EA2W group in the same brain regions(P＜0.05).AChE expression in hippocampal CA1 area showed the similar tendency.These results showed that after CCI,the activity of AChE was down-regulated in hypothalamus and hippocampus,EA of ST36-GB34 can increase the activity of ACHE,and induce a cumulative increase;OVX-induced memory impairment(estrin level-decrease) attenuates EA-induced effects,in spite of still existing in OVX rats.
3) Effect of EA on muscarinic M_1 receptor mRNA expression in hypothalamus and hippocampus in CCI rats
After CCI,M_1 receptor mRNA expression(0.39±0.25) in hypothalamus was considerably lower than that of normal control group(0.83±0.40, P＜0.05).Compared with CCI control group(0.39±0.25) and CCI+EA2D group (0.59±0.21),M_1 receptor mRNA expression of CCI+EA2W group(0.88±0.16) was increased significantly(P＜0.05).In OVX+CCI model,M_1 receptor mRNA expression of OVX+CCI control group(0.29±0.11) was markedly lower than that of normal control group(0.83±0.40,P＜0.05).Compared with OVX+CCI (0.29±0.11) and OVX+CCI+EA2D group(0.41±0.22),the expression of M_1 receptor mRNA expression(0.76±0.35) in OVX+CCI+EA2W group was markedly increased(P＜0.05),suggesting a cumulative increase of M_1 receptor mRNA expression after EA treatment.The expression of M_1 receptor mRNA in hippocampus tissue had a similar trend in comparison with hypothalamus tissue.The expression of M_1 receptor mRNA in OVX+CCI+EA2W group (1.00±0.20) was lower than that of CCI+EA2W group (1.37±0.37),suggesting an attenuation of EA-induced cumulative effect in up-regulating M_1 receptor mRNA expression after memory impairment in OVX rats.
4) Effect of EA on PKA activity in hypothalamus and hippocampus in CCI rats
Results of immunohistochemistry showed that the expressions of PKA of PVN(99.28±19.33,integral grey value),ARC(100.18±14.52) and SON (103.26±17.12) of hypothalamus in CCI+EA2W group were significantly higher than those in PVN(133.54±6.31),ARC(127.84±18.91) and SON (133.53±21.34) in normal control group;and PVN(124.35±6.3),ARC (121.77±14.66),SON(118.61±9.64) in CCI+2D group(P＜0.05).PKA expressions of PVN(107.48±12.39),ARC(117.11±19.87) and SON (129.12±16.96) in OVX+CCI+EA2W group were also markedly higher than PVN (145.96±8.02),ARC(134.24±18.43) and SON(148.42±10.7,P＜0.05) in OVX+CCI group,displaying upregulation and cumulative increase of PKA expression after EA2W.PKA expression of PKA of ARC and SON in OVX+CCI+EA2W group was markedly lower than that in CCI+EA2W group(P＜0.05),suggesting an apparent attenuation of EA-induced cumulative upregulation effect of PKA expression in OVX rats.The changing tendency of PKA expression in hippocampal CA1 region was similar to that of hypothalamic SON after repeated EA of ST36-GB34 in CCI and OVX+CCI rats.
Conclusion
1) EA of "Zusanli"(ST36)-"Yanglingquan"(GB34) is able to significantly alleviate pain sensitivity caused by CCI,and has a cumulative effect,which is weakened obviously after neuron-memory injury.
2) EA can significantly reduce CCI-induced inhibition of the expression of ACHE,M_1 receptor mRNA in hypothalamus and hippocampus,and has an accumulative upregulation effect on ACHE,PKA and M_1 receptor mRNA expression,which are closely associated with the animals' neuron-memory ability.
Therefore,there is a close relationship between the accumulative analgesic effect of EA and its resultant increase of the expression of ACHE,M_1 receptor mRNA and PKA in hypothalamus and hippocampus,and the function of neuron-memory is involved in the cumulative effect of repeated EA treatment.Results of the present study provide a set of detailed experimental evidence for repeated acupuncture-induced cumulative effect in the treatment of chronic pain.
引文
[1]蔡荣林,胡玲,周逸平等.电针对急性心肌缺血家兔心功能及心交感神经电活动的影响。针刺研究.2007,32(4):243-246.
[2]王新宇,石现,何磊.电针对急性胰腺炎患者胃肠动力的影响及其机制研究.针刺研究,2007,32(3):199-202.
[3]Megan Long.Acupuncture and medicine:promoting the body' s natural ability to heal itself.(http:/www.michiganacupuncture.org/faq/html)
[4]党文,杨介宾.针刺治疗胃癌痈的临床研究.中国针灸,1995年增刊:41-46.
[5]Capodice JL,Jin Z,Bemis DL,Samadi D,Stone BA,Kapan S,Katz AE.A pilot study on acupuncture for lower urinary tract symptoms related to chronic prostatitis/chronic pelvic pain。 Chin Med.2007,6;2:1-7.(http://www.cmjournal.org/content/2/1/1).
[6]苏静,沈素娥.针刺治疗单纯性肥胖32例.南京中医药大学学报,2005,21(4):262-263.
[7]高洁.针刺治疗肌收缩性头痛41例.辽宁中医学院学报,2005,7(5):498.
[8]杨俊生,杨素英,许武定.针刺治疗脑血管病所致假性球麻痹96例观察.针灸临床杂志,1999,15(5):9-11.
[9]夏春发,黄丽萍,刘国强.针刺背俞穴治疗老年性便秘45例.陕西中医,2006,27(1):95-96.
[10]王传年.烧山火浅刺透刺治疗周围型面神经麻痹疗效观察.四川中医。2007,25(3):104-105.
[11]孙毓,李伟,张志刚,赵素杰.电针分型预测结合面神经肌电图治疗周围性面神经麻痹80例.中国临床康复。2006,10(3):21-23.
[12]卢峻,时宇静,金智秀,图娅.不同频率电针对模型大鼠抗抑郁效应的比较研究.2003,26(6):83-85
[13]Takaoka Y,Ohta M,Ito A,Takamatsu K,Sugano A,Funakoshi K,Takaoka N,Sato N,Yokozaki H,Arizono N,Goto S,Maeda E.Electroacupuncture Suppresses Myostatin Gene Expression:Cell Proliferative Reaction in Mouse Skeletal Muscle.Physiol Genomics.2007 Jul 18;30(2):102-10.
[14]陈英辉,黄显奋.累加电针对脑缺血大鼠皮层脑源性神经营养因子表达及脑梗塞体积的影响.针刺研究,2000,25(3):165-169.
[15]Jong MS,Hwang SJ,Chen FPEffects of electro-acupuncture on serum cytokine level and peripheral blood lymphocyte subpopulation at immune-related and non-immune-related points.Acupunct Electrother Res.2006;31(1-2):45-59
[16]窦思东,刘凯,方小玲,王永中.针刺疗程长短与刺激量对针灸效应的影响.福建中医学院学报,1999,9(1):27-28.
[17]吴鎏桢,崔彩莲,韩济生.多次电针刺激抑制大鼠吗啡戒断症状的累加效应及长时程后效应.中国疼痛医学杂志,2001,7(2):105-108.
[18]朱庆丰.电针刺激对大鼠吗啡依赖的影响研究.安庆师范学院学报.2007,13(1):98-100.
[19]王宁华.穴位TENS对正常老年人热痛阈的累加抑制作用.中华物理医学与康复杂志,2003,25(9):533-536.
[20]张玉英,栾述智.口针疗法治疗坐骨神经痛45例.中国民间疗法,2001,9(4):16.
[21]聂发华.电针治疗坐骨神经痛50例.新疆中医,1999,66(2):25.
[22]王贺春,万有,姚磊,韩济生.不同频度电针治疗大鼠慢性神经源性痛的疗效比较.针刺研究,2002,27(2):112-118.
[23]Yim YK,Lee H,Hong KE,Kim YI,Lee BR,Son CG,Kim JE.Electro-acupuncture at acupoint ST36 reduces inflammation and regulates immune activity in Collagen-Induced Arthritic Mice.Evid Based Complement Alternat Med.2007Mar;4(1):51-57.
[24]杨向红,王彦青,高秀,吴根诚.电针对大鼠神经痛痛敏分数的影响.针刺研究,2002,27(1):60-63.
[25]董志强,马飞,笪翠娣,谢虹,李为民,王彦青,吴根诚.累加电针提高神经痛大鼠背根神经节GDNFmRNA的表达.上海针灸杂志,2005,24(2):33-36.
[26]张文光,董卫国,宋丹云,任君宏,林岚.针刺对大鼠坐骨神经损伤修复的影响.福建中医学院学报,2006,16(1):21-23.
[27]李为民,崔可密,吴根诚.电针治疗肠易激综合征大鼠作用观察.上海针灸杂志,2006,25(3):43-47.
[28]罗非,李楠,田津斌,等.反复电针对单发佐剂关节炎模型大鼠脊髓物质八肽胆囊收缩素及甲硫氨酸脑啡肽样免疫活性物质自发释放的影响.中国疼痛医学杂志,1996,2 (3):169-175.
[29]Han Z,Jiang YH,Wan Y,et al,Endomorphin-1 mediates 2Hz but not 100Hz electroacupuncture analgesia in the rat.[J].Neurosci Lett,1999,274(2):75-78.
[30]孙瑞卿,王贺春,王韵,罗非,韩济生.不同频率的电针对大鼠神经源性痛的治疗作用.中国应用生理学杂志,2002;18(2):128-131.
[31]Sandkuhler J,Chen JG,Cheng G,et al.Low-frequency stimulation of afferent A δ -fiber indeces long term depression at primary afferent synapses with substantia gelatinosa neurons in the rat.J Neurosic,1997,17:6483-6491.
[32]刘红香.多次100HzTENS治疗慢性炎症痛的适宜参数和可能的神经化学机制研究.生理科学进展。1999,30(1):35-37。
[33]Baffi JS,PalkovitsM.Fine topography of brain areas activated by cold stress.A fos immunohistochemical study in rats.Neuroendocrinology,2000,72:102-113.
[34]宋春杰,齐建国,章为.束缚应激大鼠下丘脑内一氧化氮合酶与c-fos蛋白的共存.神经解剖学杂志,1997,13:161-166.
[35]万顺伦,孙锦平,李召峰.应激后大鼠下丘脑磷酸化p38MAPK的变化及电针足三里穴的调节作用.中华物理医学与康复杂志,2005,27(4):202-204.
[36]Herman JP,Cullinan WE.Neurocircuitry of stress:central control of the hypothalamopituitary-adrenocortical axis.Trends Neurosci,1997,20:78-84.
[37]Herman JP,Dolgas CM,Carlson SL.Ventral subiculum regulates hypothalamo-pituitary-adrenocortical and behavioral responses to cognitive stressors.Neuroscience,1998,86:449-459.
[38]柯青,王廷华,赵有成.针刺镇痛及其机理.四川解剖学杂志,2002,10(4):225-230.
[39]杜俊辉,王妙珍,何莲芳.针刺时下丘脑弓状核区前阿黑皮原及C-fos基因的表达.针刺研究,1994,19(Z1):39.
[40]印其章,端木肇夏,俞光弟等.下丘脑弓状核、中缝背核利蓝斑在针刺镇痛中的作用及其相互关系.医学研究杂志,1991,20(6):27-28.
[41]俞昌喜,吴根诚,徐绍芬等.褪黑素减弱大鼠下丘脑弓状核β-内啡肽免疫反应的强度.Acta Physiologica SiniCa.2000,52(3):263-266.
[42]陈泽斌,熊迎春,吴吕清,袁尚朵.手针刺激大鼠“肾俞”穴对下丘脑室旁核功能状态的影响.湖北中医学院学报,1999,1(1):30-32.
[43]陈易风,汪一汗,印其章.大鼠下丘脑室旁核在针刺镇痛中的作用.针刺研究,1990,(1):32-38.
[44]杨俊,宋朝佑,林葆城,王成海,刘文彦.下丘脑室旁核参与电针镇痛.针刺研究,1989,14(1-2).111-112
[45]汉永勤,印其章.大鼠下丘脑室旁核参与针刺镇痛的电生理学观察.针刺研究,1992,3:161-165.
[46]杨俊,宋朝佑,林葆城,朱鹤年.刺激和损毁下丘脑室旁核对针刺镇痛的影响。生理学报,1992,44(5),455-460.
[47]舒志俊、杨虎川、刘静良、胡庆利、王芙蓉.家兔电针麻醉时下丘脑视上核分泌物动态变化.宁夏医学院学报,1980,3:1-6.
[48]青岛医学院针麻组,下丘脑视上核在电针镇痛中作用的初步观察.针刺研究,1978,3(1):78-79.
[49]林葆城,宋朝佑,祝元祥,刘真,吴崇仁,朱鹤年,下丘脑与针刺镇痛关系的研究,生理科学,1983,3(1):11。
[50]李玉娟,彭书崚,万朝权,曹林,李艳萍.慢性疼痛对幼鼠学习记忆行为及海马Bcl-2mRNA、脑源性生长因子mRNA的影响.中华儿科杂志,2005,43(6):444-448.
[51]彭书崚,李玉娟,万朝权,王寿平,李珏.慢性炎性疼痛对新生大鼠学习记忆功能发育及海马前脑啡肽mRNA表达的影响.中华麻醉学杂志,2005,25(8):601-603.
[52]中医研究院针灸经络研究所.针刺过程中下丘脑、海马、纹状体内单胺类神经介质及其代谢产物含量的变化。针刺研究,1978,(1):64-65.
[53]汤慈美,魏景汉,陈双双,刘善纯,林文娟.海马在电针镇痛中作用的初步探讨.针刺研究,1979.01:58-61.
[54]Lou Z,Sun W,Liu Y,Tong Z.Effect of electro-acupuncture on cortical and hippocampal EEG in adjuvantarthritisrats。Zhen Ci Yan Jiu.,1992,17(2):129-32。
[55]曾玲,卢树斌,谢惠明等.刺激体感Ⅰ区对海马痛单位放电的影响及其与电针的关系.广西医科大学学报,1994,11(1):31-34.
[56]柯青,王廷华,赵有成.针刺镇痛及其机理.四川解剖学杂志,2002,10(4):225-230.
[57]Li HL,Li XC,Ruan HZ,Liu ZZ.Relationship between the content of central substance P and the analgesic effect of electroacupuncture in rats。 Zhen Ci Yan Jiu.1989;14(3):370-4。
[58]许建刚,冯琼,赵仓焕,宋开源.电针对实验性类风湿性关节炎海马中C-fos表达的影响.中国针灸,1998,(1):47-49.
[59]王彤,于建春,邢海涛,熊会海,韩景献.“益气凋血,扶本培元”针刺法对多发梗塞性痴呆大鼠海马bcl-2、bax蛋白表达的影响.上海中医药大学学报,2006,20(3):58-60.
[60]王琴玉,孙砚辉,许能贵,靳瑞.不同时窗针刺对脑瘫幼鼠海马CA1区神经元及脑组织神经生长因子表达的影响。针刺研究,2004,29(3):174-178.
[61]刘存志,于建春,张雪竹,王彤,韩景献.多发梗塞性痴呆大鼠海马抗氧化酶活性的变化及针刺的干预作用.中华中医药杂志(原中国医药学报),2005,20(12):724-726.
[62]丁晓蓉,于建春,于涛,韩景献.利用基因芯片研究针刺对快速老化小鼠海马基因表达的影响.天津中医药,2005,22(5):394-398.
[63]罗世杰,白学斌,苗晋,邓春雷,田志丽,苏保宁,宋启劳,张卉,苗琦.益智合剂及针刺百会影响幼龄大鼠智力、海马超微结构及NO含量的实验研究.四川中医,2006,24(1):23-24.
[64]Huang ZN,Yang R,Chen G,Cheng JS.Effect of electroacupuncture and 7-NI on penicillin-induced epilepsy and their relation with intrahippocampal NO changes。Sheng Li Xue Bao.1999 0ct;51(5):508-14.
[65]王世军,崔可密,卢岩,郭继龙,韩冰.针刺对MCAO大鼠海马CA3区微血管数日及神经元死亡率的影响.山东中医药大学学报,2005,29(1):50-60.
[66]姜桂美,贾超,赖新生.针刺对阿尔茨海默病模型大鼠神经细胞凋亡的影响.上海针灸杂志,2006,25(12):33-36.
[67]原淑娟,张忐雄,邱虹等.电针对D-半乳糖所致衰老模型大鼠神经细胞凋亡的影响.中国临床康复,2005,9(25):132-134.
[683 Davies P,Maloney AJ.Selective loss of contral cholinergic neurons in Alzheimer ' s disease[J].Lancet,1976,25;2(8000):1403。
[69]Bartus RT,Dean RL,Beer B,etal.The cholinergic hypothesis of geriatric memory dysfunction[J].Science,1982,30;217(4558):408-414.
[70]Haba K,Ogawa N,Mizukawa K,et al.Time course of changes in lipid peroxidation,pre- and postSYNaptic cholinergic indices,NMDA receptor binding and neuronal death inthegerbilhippocampusfollowingtransientischemia.BrainRes,1991;540(1-2):116-122
[71]Hefti F,Dravid A,Hartikka J.Chronic intraventricular injections of nerve growth factor elevate hippocampal choline acetyltransferase activity in adult rats with partial septo-hippocampal lesions[J].Brain Res,1984,293(2):305-311.
[72]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:176.
[73]董洪涛,白英。乙酰胆碱多因素阿尔茨海默病动物模型的建立及针刺对其海马胆碱酯酶活性影响的实验研究。上海医学,2002,25(7):427-429。
[74]Muller WE,Stoll L,Schubert T,et al·Central cholinergic function-ing and aging [J]· Acta Psychiatr Scand SuPPl,1991;366:34-9 ·
[75]Bonkale WL,Fastbom J,Wiehager B,et al·Impaired G-protein-stimulated adenylyl cyclase activity in Alzheimer' s disease brain is not accompanied by reduced cyclic-AMP-dependent protein kinase Aactivity.(J) Brain Res,1996;737(1-2):155-61 ·
[76]Flynn DD,Ferrari-DiLeo G,Levey AI,et al · Differential alterations in muscarinic receptor subtypes in Alzheimer' s disease:Implications for cholinergic-based therapies[J]· Life Sci· 1995;56(11-12):869-7.
[77]Nordberg A,A1afuzoff I,Winblad B· Nicotinic and muscarinic sub-types in the human brain:Changes with aging and dementia[J]· J Neurosci Res,1992;31(1):103-10 ·
[78]Ferreira AR,FHrstenau L,Blanco C,Kornisiuk E,Sanchez G,Daroit D,Castro e Silva M,Cervenansky C,Jerusalinsky D,Quillfeldt JA.Role of hippocampal M1 and M4 muscarinic receptor subtypes in memory consolidation in the rat.Pharmacol Biochem Behav.2003,74(2):411-415.
[79]Ghelardini C,Galeotti N,Matucci R,Bellucci C,Gualtieri F,Capaccioli S,Quattrone A,Bartolini A.Antisense 'knockdowns' of M1 receptors induces transient anterograde amnesia in mice.Ueuropharmacology.1999 Mar;38(3):339-48.
[80]Miyakawa T,WamadaM,DuttaroyA,WessJ.Hyperactivityandintact hippocampus-dependent learning in mice lacking the M1 muscarinic acetylcholine receptor.J Neurosci.2001Jul 15;21(14):5239-50.
[81]黄忠致等.猕猴体感Ⅰ区皮层内注射甲基硫酸新斯的明对痛阈的影响。第十九届中国生理学大会学术论文摘要汇编。生理通讯增刊,1994:112.
[82]徐嵘,关新民,王才源.电针及辣椒素处理对脊髓后角ACHE活性和[3H]-QNB结合位点的影响.针刺研究,1996,21(2):47-51.
[83]关新民,施静,张育文.针刺痛时脊髓后和脊神经节内Ach代谢的变化.针刺研究,1990,15(1):18-23。
[84]关新民,王才源,余福.中枢乙酰胆碱与针刺镇痛关系的研究.针刺研究,1991,16(2):129-137.
[85]魏月娥,傅冠英,万选才.针刺后交感神经中枢内神经介质变化的组织化学观察.解剖学报,1983,14(2):185.
[86]徐维,阎亚生,石宏,陈正秋.乙酸胆碱参与皮层下行调制疼痛的作用.针刺研究,1991,16(3):155-156.
[87]潘玉贞,吴建群,王绍.乙酰胆碱(ACh)兴奋外侧缰核拮抗电针镇痛.长春中医学院学报,1997.13(61).58-59
[88]Lewis JW,Cannon JT,Liebeskind JC.Involvement of central muscarinic cholinergic mechanisms in opioid stress analgesia.Brain Res.1983,270(2):289-93.
[89]Katayama Yet al:Brain Res,296:263.1984.Katayama Y,Watkins LR,Becket DP,Hayes RL.Non-opiate analgesia induced by carbachol microinjection into the pontine parabrachial region of the cat.Brain Res.1984:296(2):263-83.
[90]张立新,刘祚延.微电泳乙酰胆碱和阿托品对大鼠丘脑束旁核痛敏神经元电活动的影响.生理学报,1993,45(1):55-61.
[91]孙明智,范平,张季叶等.乙酰胆碱对丘脑束旁核痛兴奋和痛抑制神经元放电的影响及其机制的探讨,针刺研究,1989,14(1-2):104-105.
[92]徐广银,等.内源性胆碱能系统在佐剂性关节炎大鼠电针镇痛中的作用。第十九届中国生理学大会学术论文摘要汇编,生理通讯增刊,1994,112.
[93]Smith MD,Yang XH,Nha JY,Buccafusco JJ.Antinociceptive effect of spinal cholinergic stimulation:interaction with substance 。 Life Sci.1989;45(14):1255-61。
[94]Hamm RJ,Knisely JS.Developmental differences in the analgesia produced by the central cholinergic system.Dev Psychobiol.1987 May;20(3):345-54.
[1]Megan Long.Acupuncture and medicine:promoting the body' s natural ability to heal itself.(http:/www.michiganacupuncture.org/faq/html)
[2]韩济生.影响针刺镇痛效果的若干因素.针刺研究,1994,19(3-4):1-4.
[3]董志强,马飞,笪翠娣,谢虹,李为民,王彦青,吴根诚.累加电针提高神经痛大鼠背根神经节GDNFmRNA的表达.上海针灸杂志,2005,24(2):33-36.
[4]孙瑞卿,王贺春,景峥,王韵,万有,韩济生.2Hz电针减轻神经源性痛大鼠的痛觉超敏利冷诱发的持续性疼痛.中国疼痛医学杂志,2003,9,(4):220-224.
[5]王贺春,万有,姚磊,韩济生.不同频度电针治疗大鼠慢性神经原性同的疗效比较.针刺研究,2002,27(2):112-118.
[6]王文靖,赵仓焕,谢步霓,盛佑祥,朱小华.不同间隔时间电针对炎症痛大鼠下丘脑脑阿片肽基因表达的影响.针刺研究,2006,31(3):131-135.
[7]罗非.反复电针对慢性痛的累加治疗作用及其机制研究.生理科学进展,1996,7(3):241-244.
[8]陈瑾,刘光谱,唐勇.中枢及外周5-HT、5-HIAA在针刺镇痛后效应中的作用.中医药学刊,2003,21(9):1446-1450.
[9]Bennett GJ,Xie YK.A peripheral mononeuropathy in the rat that produces disorders of pain sensation like those seen in man.Pain,1988,33(1):87-107.
[10]聂发华.电针治疗坐骨神经痛50例.新疆中医,1999,66(2);25.
[11]陈晓红,康妹娟,赵淳,韩济生.大鼠脊髓中三类阿片受体均参与2-15Hz电针镇痛.北京医科大学学报,1992,24(1):54.
[12]刘俊岭,陈淑萍,高永辉,孟凡颖.不同强度、不同频度电针对慢性痛大鼠镇痛作用的比较.针刺研究,2006,31(5):280-285.
[13]王文靖,赵仓焕,谢步霓,盛佑祥,朱小华.不同间隔时间电针对炎症痛大鼠下丘脑脑阿片肽基因表达的影响.针刺研究,2006,31(3):131-135.
[14]Palmon SC,Megan J.Estrogen increases cGMP inselected brain regions and in cerbral microvessels.J Cerb Blood Flow Metab,1998,18(1):1248-1252.
[15]Behl C,Lezoualc' h F.Estrogenswith an intact phenolic group prevent death of neuronal cells following glutathione depletion.Restor Neurol Neurosci,1998,12(223):127-134.
[16]Panikck KS,Guan G,King MA.17beta-estradiol attenuates CREB decline in the rat hippocampus following seizure.J Neurobiol,1997,33(7):961-967.
[17]DianeD,Murphy,NelsonB,et al.Estradiol increasesdendriticspinedensity by reducing GABA neurotransmission in hippocampal neurons.J Neurosci,1998,18(7):2 550-2 559.
[18]Henderson VW.Estrogen,cognition and a woman's risk of Alzheimer's disease [J].Am J Med,1997,103(3A):11S-18S.
[19]Asthana S,Craft S,Baker LD,etal.Cognitive and neuroendocrine response to transdemal estrogen in postmenopauseal woman with Alzheimer's disease:results of placebo controlled,double-blind,pilot study[J].Psychoneuroendocrinology,1999,24(6):657-677.
[201Dong H T,Zhang T S,Chen Y L,etal.Survey and analysis on Alzheimer's disease treated by traditional Chinese medicine[J].Shanghai J Tradit Chin Med(上海中医药杂志),2000,20(4):17-19.
[21]Zhao LX,Chen O,Diaz Brinton,et al.Neuroprotective and neurotrophic efficacy of phytoestrogens in cultured hippocampal neurons[J].Exp Biol Med(Maywood).2002,227(7):509-519.
[22]Dubal DB,Zashon ML,Pettigrew LC,et al.Estradiol protects against ischemic injury[J].J Cereb Blood Flow Metab,1998,18(11):1253-1258.
[23]Birge SJ.Is there a role for estrogen replacement therapy in the prevention and treatment of dementia ? J Am Geriatr 8oc,1996;44(7):865-870.
[24]Singh M,Meyer EM,Simpkins JW.Ovarian steroid deprivation ersults in areversible learning impairment and compromised cholinergic function in female Sprague-Dawley rats.Brain Res,1994;644(2):305-12.
[25]NovelliG,Reichardt JKV.Molecularbasis ofdisorderofhuman galactose m etabolism:past,present,and future[J].Molecular Genetics and Metabolism,2000,71(1):62-65.
[26]Forcier NJ,Mizisin AP,Rmimer M A,et al.Cellular pa-thology of the nerve microenvironment in galactose in-toxication[J].J Neuropathol Exp Neuro,1991,50(3):236-255.
[27]陈勤.抗衰老研究实验方法[M].北京:中国医药科技出版社,1996:494-496.6.
[28]王月华,杜冠华.L-肉毒碱对D-半乳糖衰老模型小鼠的作用.中国老年学杂志, 2003,23(1):49-51.
[29]Hua X,Lei M,Zhang Y,Ding J,tlan Q,Hu G,Xiao M.Long-term D-galactose injection combined with ovariectomy serves as a new rodent model for Alzheimer' s disease.Life Sci.2007;80(20):1897-905.
[30]曲淑君.疼痛的治疗。中国临床康复,2003,6(12):1790-1.
[31]严相默。神经阻滞技术与周围神经痛.中国临床康复,2003,7(6):881-5.
[32]Zheng LF,Wang R,Xu YZ,Yi XN,Zhang JW,Zeng ZC.Calcitonin gene-related peptide dynamics in rat dorsal root ganglia and spinal cord following different sciatic nerve injuries.Brain Res 2008 2;1187:20-32.
[33]宋学军.神经病理性疼痛的动物模型和分子机制.江苏临床医学杂志,2002,6(5):360-399.
[34]赵峰,戴红,郑泓溶,等.中枢性疼痛机制的研究进展.中华物理医学与康复杂志,2003,25(5):310-320.
[35]Bennett GJ,Xie YK.A peripheral mononeuropathy in rat produce disorders of pain sensation like those seen in man.PAIN,1988;33(1):87-107.
[36]Nuytten D,Kupers R,Lamme S,et al.Further evidence for myelinated as well as unmyelinated fiber damage in a rat model of neuropathic pain.Exp Brain Res,1992:91(1):73-8.
[37]Maves TJ,Pechman PS,Gebhart GF,et al.Possible chemical contribution from chromic gut sutures produces disorders of pain sensation like those seen in man.Pain,1993:54(1):57-69.
[38]Attal N,Jazat F,Kayser V,et al.Further evidence for'pain-ralated'behaviour in a model of unilateral peripheral mononeuropathy.Pain,1990;41(2):235-51.
[39]杨向红,王彦青,高秀,吴根诚,电针对大鼠神经痛痛敏分数的影响.针刺研究,2002,27(1):60-63.
[40]董志强,马飞,笪翠娣,谢虹,李为民,王彦青,吴根诚.累加电针提高神经痛大鼠背根神经节GDNFmRNA的表达.上海针灸杂志,2005,24(2):33-36。
[41]李忠仁主编,实验针灸学,中国中医药出版社,2003年1月第一版:174.
[42]Giancar P,Giacomo S.Nootropic drugs and brain cholinergic mechanism[J].Pro Neuro Psychopharmacol and Biol Psychiat,1989,13:77.
[43]孟凡颖,屯针“足三里”-“阳陵泉”穴区治疗慢性痛累积效应机制的分析,2007,硕士论文文库。
[44]刘洁生,谢丽玲,洪岸,等.神经元烟碱乙酰胆碱受体受阻滞所致老年痴呆动物模型的建立.卫生研究,1999,28(4):32-35.
[45]Mesulam MM,Van Hoesen GW.Acetylcholinesterase-rich projections from the basal forebrain of the rhesus monkey to neocortex.Brain Res.1976 Jun 4;109(1):152-7.
[46]张艳,熊希凯.骨骼肌运动终板和神经纤维的酶组织化学及镀银双重显示.中国组织化学与细胞化学杂志,1998,(7):364.
[47]Mesulam M-M,Volicer L,Marquis JK,et al.Systematic regional differentces in the cholinergic innervation of primate cerebral cortex:distribution of enzyme activities and some behavioral implications.Ann Neurol,1986,19:144-151.
[48]Geula C,Mesulam MM.Cortical cholinergic fibers in aging and Alzheimer' s disease:a morphometric study[J].Neuroscience,1989,33:469-481.
[49]李质馨,田洪艳,鲁质博,等.胎脑提取液对坐骨神经损伤大鼠脊髓运动神经元酶活性的影响.解剖学杂志,2003,26(5):443-446.
[50]薛景凤,杨振军,李健.坐骨神经局部缺血后相关神经元胞体乙酰胆碱酯酶变化的定量分析.解剖学杂志,2007,30(1):54-56.
[51]杨军,王静,姜文,等.赤勺总甙对D-半乳糖衰老小鼠学习记忆及代谢产物的影响.中国药理学通讯,2001,17(6):697.
[52]Rodriguez-Puertas R,Pascual J,Vilaro T,et al · Autoradiographic distribution of M1,M2,M3,and M4 muscarinic receptor subtypes in Alzheimer' s disease [J]· Synapse,1997;26(4):341-50 ·
[53]Pauly JR·Nicotinic cholinergic receptor deficits in Alzheimer' s disease:Where' s the smoke[J]J Alzheimers Dis,1999:1(4,5):221-30·
[54]王才源,等.电针镇痛对太鼠脑内ACh,ChAc及AChE活性的影响.针刺研究,1987,12(2):60.
[55]关新民,施静,张育文.针刺痛时脊髓后和脊神经节内Ach代谢的变化.针刺研究,1990,15(1):18-23。
[56]林真.试析乙酰胆碱酯酶活性传变与针灸作用及经络现象的关系.中国针灸,1996,43(1):43-45.
[57]王少锦,李爱英,张雪静.针刺督脉穴对拟AD大鼠海马区Ache、ChAT活性的影响.南京中医药大学学报,2004,20(5):289-291。
[58]柏志全,郑辉.姚平.电针对大鼠脑缺血再灌注后脑组织内乙酰胆碱酯酶活性的影响.中国临床康复2004,5(4):672-673.
[59]Naguib M,Yaksh TL.Characterization of muscarinic receptor subtypes that mediate antinociception in the rat spinal cord.。 Anesth Analg.1997:85(4):847-53.
[60]Shiokawa M,Yamaguchi T,Narita M,Okutsu D,Nagumo Y,Miyoshi K,Suzuki M,Inoue T,Suzuki T.Effects of fasudil on neuropathic pain-like state in mice。 Nihon Shinkei Seishin Yakurigaku Zasshi.2007;27(4):153-9.
[61]Malmberg AB,Brandon EP,Idzerda RL,Liu H,McKnight GS,Basbaum AI.Diminished inflammation and nociceptive pain with preservation of neuropathic pain in mice with a targeted mutation of the type Ⅰ regulatory subunit of cAMP-dependent protein kinase.J Neurosci.1997,17(19):7462-70.
[62]Yajima Y,Narita M,Shimamura M,Narita M,Kubota C,Suzuki T.Differential involvement of spinal protein kinase C and protein kinase A in neuropathic and inflammatory pain in mice.Brain Res.2003,992(2):288-93.
[63]Miletic G,Pankratz MT,Miletic V.Increases in the phosphorylation of cyclic AMP response element binding protein(CREB) and decreases in the content of calcineurin accompany thermal hyperalgesia following chronic constriction injury in rats.Pain.2002,99(3):493-500.
[64]王哲,随瑜,德亮,炜熙.性应激对大鼠海马神经元PKA和P-CREB蛋白表达的影响及抗抑郁剂的拮抗作用.中南大学学报,2006,31(5):767-771.
[65]洪小平,刘能保,张敏海,等.慢性复合应激对大鼠学习和记忆功能及海马内PKA-C _β表达的影响.华中科技大学学报,2005,34(3):257-261.
[66]Liang Z,Liu F,6rundke-Iqbal I,Iqbal K,Gong CX.Down-reguiation of cAMP-dependent protein kinase by over-activated calpain in Alzheimer disease brain.J Neurochem.2007,103(6):2462-70.
[67]Tanaka Y,Takase M,6amo S.Relationship between general anesthesia and memory in Drosophila involving the cAMP/PKA pathways and adhesion-related molecules.Curr Med Chem.2007;14(13):1479-88.
[68]Takeo S,Niimura M,Miyake-Takagi K,et al.A possible mechanism for improvement by a cognition-enhancer nefiracetam of spatial memory function and c AMP - mediated signal transduction system in sustained cerebral ischaemia in rats.Br J Pharmacol 2003;138(4):642-54.
[69]Bernabeu R,Bevilaqua L,Ardenghi,et al.Involvement of hippocapal c AMP/c AMP-dependent protein kinase signaling pathways in a late memory consolidation phase of aversively motivated learning in rats.Proc Natl Acad Sci U S A 1997,94(13):7041-6.
[70]Waltereit R,Weller M.Signaling from c AMP/PKA to MAPK and synaptic plasticity.Mol Neurobiol 2003,27(1):99-106.
[71]Nagakura A,Niimura M,Takeo S.Effects of a phosphodiesterase Ⅳ inhibitor rolipram on microsphere embolism-induced defects in memory function and cerebral cyclic AMP signal transduction system in rats.Br J Pharmacol 2002,135(7):1783-93.
[72]常铉,舒斯云,包新民,江刚.PKA-CREB信号通路在大鼠纹状体边缘区及海马空间学习记忆过程中的作用.中医临床康复,2005,9(1):39-41.
[73]Smith JA,Davis CL,Burgess GM.Prostaglandin E2--induced sensitization of bradykinin--evoked responses in rat dorsal root ganglion neurons is medlated by cAMP --dependent protein kinase A.Eur J Neuorsci.2000:12(9):3250-3258.
[74]Malmber AB,Brandon EP,Idzerda RL,et al.Diminished inflammation and nociceptive pain with preservation of neuropathic pain in mice with a targeted mutation of the type Ⅰ regulatory subunit of c AMP-dependent protein kinase.J Neurosci 1997;17(19):7462-70.
[751Codere TJ.YashPal K.Intracellular messengers contributing to Persistent nociception and hyperalgesia induced by-Lglutamate and substance P in the rat formalin pain model.Eur J Neuorsci.1994;6(8):1328--1334.
[76]姚永兴,张励才,宋学军,曾冈明.鞘内注射H-89对神经病理痛大鼠脊髓背角磷酸化cAMP 反应元件结合蛋白表达的影响.中华麻醉学杂志,2005,25(5):371-374.
[77]Shiokawa M,Yamaguchi T,Narita M,Okutsu D,Nagumo Y,Miyoshi K,Suzuki M,Inoue T,Suzuki TEffects of fasudil on neuropathic pain-like state in mice.Nihon Shinkei Seishin Yakurigaku Zasshi.2007 Aug;27(4):153-9.
[78]陆汎、朱红梅、谢俊杰,周海虹,陈艳玲,胡纪原。屯针对抑郁症模型大鼠行为学、血浆COR及海马PKA、PKC表达的影响。中国针灸,2008,28(3):214-8.
[79]张敏,图娅.电针对慢性应激抑郁模型大鼠学习记忆行为的影响.中医研究,2008,21(3):15-18。
[80]付弋,梁繁荣,陶巧林.不同针法对佐刑性关节炎大鼠下丘脑镇痛效应比较.上海针灸杂志,2005,24(5):38-41.
[2]王新宇,石现,何磊.电针对急性胰腺炎患者胃肠动力的影响及其机制研究.针刺研究,2007,32(3):199-202.
[3]Megan Long.Acupuncture and medicine:promoting the body' s natural ability to heal itself.(http:/www.michiganacupuncture.org/faq/html)
[4]党文,杨介宾.针刺治疗胃癌痈的临床研究.中国针灸,1995年增刊:41-46.
[5]Capodice JL,Jin Z,Bemis DL,Samadi D,Stone BA,Kapan S,Katz AE.A pilot study on acupuncture for lower urinary tract symptoms related to chronic prostatitis/chronic pelvic pain。 Chin Med.2007,6;2:1-7.(http://www.cmjournal.org/content/2/1/1).
[6]苏静,沈素娥.针刺治疗单纯性肥胖32例.南京中医药大学学报,2005,21(4):262-263.
[7]高洁.针刺治疗肌收缩性头痛41例.辽宁中医学院学报,2005,7(5):498.
[8]杨俊生,杨素英,许武定.针刺治疗脑血管病所致假性球麻痹96例观察.针灸临床杂志,1999,15(5):9-11.
[9]夏春发,黄丽萍,刘国强.针刺背俞穴治疗老年性便秘45例.陕西中医,2006,27(1):95-96.
[10]王传年.烧山火浅刺透刺治疗周围型面神经麻痹疗效观察.四川中医。2007,25(3):104-105.
[11]孙毓,李伟,张志刚,赵素杰.电针分型预测结合面神经肌电图治疗周围性面神经麻痹80例.中国临床康复。2006,10(3):21-23.
[12]卢峻,时宇静,金智秀,图娅.不同频率电针对模型大鼠抗抑郁效应的比较研究.2003,26(6):83-85
[13]Takaoka Y,Ohta M,Ito A,Takamatsu K,Sugano A,Funakoshi K,Takaoka N,Sato N,Yokozaki H,Arizono N,Goto S,Maeda E.Electroacupuncture Suppresses Myostatin Gene Expression:Cell Proliferative Reaction in Mouse Skeletal Muscle.Physiol Genomics.2007 Jul 18;30(2):102-10.
[14]陈英辉,黄显奋.累加电针对脑缺血大鼠皮层脑源性神经营养因子表达及脑梗塞体积的影响.针刺研究,2000,25(3):165-169.
[15]Jong MS,Hwang SJ,Chen FPEffects of electro-acupuncture on serum cytokine level and peripheral blood lymphocyte subpopulation at immune-related and non-immune-related points.Acupunct Electrother Res.2006;31(1-2):45-59
[16]窦思东,刘凯,方小玲,王永中.针刺疗程长短与刺激量对针灸效应的影响.福建中医学院学报,1999,9(1):27-28.
[17]吴鎏桢,崔彩莲,韩济生.多次电针刺激抑制大鼠吗啡戒断症状的累加效应及长时程后效应.中国疼痛医学杂志,2001,7(2):105-108.
[18]朱庆丰.电针刺激对大鼠吗啡依赖的影响研究.安庆师范学院学报.2007,13(1):98-100.
[19]王宁华.穴位TENS对正常老年人热痛阈的累加抑制作用.中华物理医学与康复杂志,2003,25(9):533-536.
[20]张玉英,栾述智.口针疗法治疗坐骨神经痛45例.中国民间疗法,2001,9(4):16.
[21]聂发华.电针治疗坐骨神经痛50例.新疆中医,1999,66(2):25.
[22]王贺春,万有,姚磊,韩济生.不同频度电针治疗大鼠慢性神经源性痛的疗效比较.针刺研究,2002,27(2):112-118.
[23]Yim YK,Lee H,Hong KE,Kim YI,Lee BR,Son CG,Kim JE.Electro-acupuncture at acupoint ST36 reduces inflammation and regulates immune activity in Collagen-Induced Arthritic Mice.Evid Based Complement Alternat Med.2007Mar;4(1):51-57.
[24]杨向红,王彦青,高秀,吴根诚.电针对大鼠神经痛痛敏分数的影响.针刺研究,2002,27(1):60-63.
[25]董志强,马飞,笪翠娣,谢虹,李为民,王彦青,吴根诚.累加电针提高神经痛大鼠背根神经节GDNFmRNA的表达.上海针灸杂志,2005,24(2):33-36.
[26]张文光,董卫国,宋丹云,任君宏,林岚.针刺对大鼠坐骨神经损伤修复的影响.福建中医学院学报,2006,16(1):21-23.
[27]李为民,崔可密,吴根诚.电针治疗肠易激综合征大鼠作用观察.上海针灸杂志,2006,25(3):43-47.
[28]罗非,李楠,田津斌,等.反复电针对单发佐剂关节炎模型大鼠脊髓物质八肽胆囊收缩素及甲硫氨酸脑啡肽样免疫活性物质自发释放的影响.中国疼痛医学杂志,1996,2 (3):169-175.
[29]Han Z,Jiang YH,Wan Y,et al,Endomorphin-1 mediates 2Hz but not 100Hz electroacupuncture analgesia in the rat.[J].Neurosci Lett,1999,274(2):75-78.
[30]孙瑞卿,王贺春,王韵,罗非,韩济生.不同频率的电针对大鼠神经源性痛的治疗作用.中国应用生理学杂志,2002;18(2):128-131.
[31]Sandkuhler J,Chen JG,Cheng G,et al.Low-frequency stimulation of afferent A δ -fiber indeces long term depression at primary afferent synapses with substantia gelatinosa neurons in the rat.J Neurosic,1997,17:6483-6491.
[32]刘红香.多次100HzTENS治疗慢性炎症痛的适宜参数和可能的神经化学机制研究.生理科学进展。1999,30(1):35-37。
[33]Baffi JS,PalkovitsM.Fine topography of brain areas activated by cold stress.A fos immunohistochemical study in rats.Neuroendocrinology,2000,72:102-113.
[34]宋春杰,齐建国,章为.束缚应激大鼠下丘脑内一氧化氮合酶与c-fos蛋白的共存.神经解剖学杂志,1997,13:161-166.
[35]万顺伦,孙锦平,李召峰.应激后大鼠下丘脑磷酸化p38MAPK的变化及电针足三里穴的调节作用.中华物理医学与康复杂志,2005,27(4):202-204.
[36]Herman JP,Cullinan WE.Neurocircuitry of stress:central control of the hypothalamopituitary-adrenocortical axis.Trends Neurosci,1997,20:78-84.
[37]Herman JP,Dolgas CM,Carlson SL.Ventral subiculum regulates hypothalamo-pituitary-adrenocortical and behavioral responses to cognitive stressors.Neuroscience,1998,86:449-459.
[38]柯青,王廷华,赵有成.针刺镇痛及其机理.四川解剖学杂志,2002,10(4):225-230.
[39]杜俊辉,王妙珍,何莲芳.针刺时下丘脑弓状核区前阿黑皮原及C-fos基因的表达.针刺研究,1994,19(Z1):39.
[40]印其章,端木肇夏,俞光弟等.下丘脑弓状核、中缝背核利蓝斑在针刺镇痛中的作用及其相互关系.医学研究杂志,1991,20(6):27-28.
[41]俞昌喜,吴根诚,徐绍芬等.褪黑素减弱大鼠下丘脑弓状核β-内啡肽免疫反应的强度.Acta Physiologica SiniCa.2000,52(3):263-266.
[42]陈泽斌,熊迎春,吴吕清,袁尚朵.手针刺激大鼠“肾俞”穴对下丘脑室旁核功能状态的影响.湖北中医学院学报,1999,1(1):30-32.
[43]陈易风,汪一汗,印其章.大鼠下丘脑室旁核在针刺镇痛中的作用.针刺研究,1990,(1):32-38.
[44]杨俊,宋朝佑,林葆城,王成海,刘文彦.下丘脑室旁核参与电针镇痛.针刺研究,1989,14(1-2).111-112
[45]汉永勤,印其章.大鼠下丘脑室旁核参与针刺镇痛的电生理学观察.针刺研究,1992,3:161-165.
[46]杨俊,宋朝佑,林葆城,朱鹤年.刺激和损毁下丘脑室旁核对针刺镇痛的影响。生理学报,1992,44(5),455-460.
[47]舒志俊、杨虎川、刘静良、胡庆利、王芙蓉.家兔电针麻醉时下丘脑视上核分泌物动态变化.宁夏医学院学报,1980,3:1-6.
[48]青岛医学院针麻组,下丘脑视上核在电针镇痛中作用的初步观察.针刺研究,1978,3(1):78-79.
[49]林葆城,宋朝佑,祝元祥,刘真,吴崇仁,朱鹤年,下丘脑与针刺镇痛关系的研究,生理科学,1983,3(1):11。
[50]李玉娟,彭书崚,万朝权,曹林,李艳萍.慢性疼痛对幼鼠学习记忆行为及海马Bcl-2mRNA、脑源性生长因子mRNA的影响.中华儿科杂志,2005,43(6):444-448.
[51]彭书崚,李玉娟,万朝权,王寿平,李珏.慢性炎性疼痛对新生大鼠学习记忆功能发育及海马前脑啡肽mRNA表达的影响.中华麻醉学杂志,2005,25(8):601-603.
[52]中医研究院针灸经络研究所.针刺过程中下丘脑、海马、纹状体内单胺类神经介质及其代谢产物含量的变化。针刺研究,1978,(1):64-65.
[53]汤慈美,魏景汉,陈双双,刘善纯,林文娟.海马在电针镇痛中作用的初步探讨.针刺研究,1979.01:58-61.
[54]Lou Z,Sun W,Liu Y,Tong Z.Effect of electro-acupuncture on cortical and hippocampal EEG in adjuvantarthritisrats。Zhen Ci Yan Jiu.,1992,17(2):129-32。
[55]曾玲,卢树斌,谢惠明等.刺激体感Ⅰ区对海马痛单位放电的影响及其与电针的关系.广西医科大学学报,1994,11(1):31-34.
[56]柯青,王廷华,赵有成.针刺镇痛及其机理.四川解剖学杂志,2002,10(4):225-230.
[57]Li HL,Li XC,Ruan HZ,Liu ZZ.Relationship between the content of central substance P and the analgesic effect of electroacupuncture in rats。 Zhen Ci Yan Jiu.1989;14(3):370-4。
[58]许建刚,冯琼,赵仓焕,宋开源.电针对实验性类风湿性关节炎海马中C-fos表达的影响.中国针灸,1998,(1):47-49.
[59]王彤,于建春,邢海涛,熊会海,韩景献.“益气凋血,扶本培元”针刺法对多发梗塞性痴呆大鼠海马bcl-2、bax蛋白表达的影响.上海中医药大学学报,2006,20(3):58-60.
[60]王琴玉,孙砚辉,许能贵,靳瑞.不同时窗针刺对脑瘫幼鼠海马CA1区神经元及脑组织神经生长因子表达的影响。针刺研究,2004,29(3):174-178.
[61]刘存志,于建春,张雪竹,王彤,韩景献.多发梗塞性痴呆大鼠海马抗氧化酶活性的变化及针刺的干预作用.中华中医药杂志(原中国医药学报),2005,20(12):724-726.
[62]丁晓蓉,于建春,于涛,韩景献.利用基因芯片研究针刺对快速老化小鼠海马基因表达的影响.天津中医药,2005,22(5):394-398.
[63]罗世杰,白学斌,苗晋,邓春雷,田志丽,苏保宁,宋启劳,张卉,苗琦.益智合剂及针刺百会影响幼龄大鼠智力、海马超微结构及NO含量的实验研究.四川中医,2006,24(1):23-24.
[64]Huang ZN,Yang R,Chen G,Cheng JS.Effect of electroacupuncture and 7-NI on penicillin-induced epilepsy and their relation with intrahippocampal NO changes。Sheng Li Xue Bao.1999 0ct;51(5):508-14.
[65]王世军,崔可密,卢岩,郭继龙,韩冰.针刺对MCAO大鼠海马CA3区微血管数日及神经元死亡率的影响.山东中医药大学学报,2005,29(1):50-60.
[66]姜桂美,贾超,赖新生.针刺对阿尔茨海默病模型大鼠神经细胞凋亡的影响.上海针灸杂志,2006,25(12):33-36.
[67]原淑娟,张忐雄,邱虹等.电针对D-半乳糖所致衰老模型大鼠神经细胞凋亡的影响.中国临床康复,2005,9(25):132-134.
[683 Davies P,Maloney AJ.Selective loss of contral cholinergic neurons in Alzheimer ' s disease[J].Lancet,1976,25;2(8000):1403。
[69]Bartus RT,Dean RL,Beer B,etal.The cholinergic hypothesis of geriatric memory dysfunction[J].Science,1982,30;217(4558):408-414.
[70]Haba K,Ogawa N,Mizukawa K,et al.Time course of changes in lipid peroxidation,pre- and postSYNaptic cholinergic indices,NMDA receptor binding and neuronal death inthegerbilhippocampusfollowingtransientischemia.BrainRes,1991;540(1-2):116-122
[71]Hefti F,Dravid A,Hartikka J.Chronic intraventricular injections of nerve growth factor elevate hippocampal choline acetyltransferase activity in adult rats with partial septo-hippocampal lesions[J].Brain Res,1984,293(2):305-311.
[72]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:176.
[73]董洪涛,白英。乙酰胆碱多因素阿尔茨海默病动物模型的建立及针刺对其海马胆碱酯酶活性影响的实验研究。上海医学,2002,25(7):427-429。
[74]Muller WE,Stoll L,Schubert T,et al·Central cholinergic function-ing and aging [J]· Acta Psychiatr Scand SuPPl,1991;366:34-9 ·
[75]Bonkale WL,Fastbom J,Wiehager B,et al·Impaired G-protein-stimulated adenylyl cyclase activity in Alzheimer' s disease brain is not accompanied by reduced cyclic-AMP-dependent protein kinase Aactivity.(J) Brain Res,1996;737(1-2):155-61 ·
[76]Flynn DD,Ferrari-DiLeo G,Levey AI,et al · Differential alterations in muscarinic receptor subtypes in Alzheimer' s disease:Implications for cholinergic-based therapies[J]· Life Sci· 1995;56(11-12):869-7.
[77]Nordberg A,A1afuzoff I,Winblad B· Nicotinic and muscarinic sub-types in the human brain:Changes with aging and dementia[J]· J Neurosci Res,1992;31(1):103-10 ·
[78]Ferreira AR,FHrstenau L,Blanco C,Kornisiuk E,Sanchez G,Daroit D,Castro e Silva M,Cervenansky C,Jerusalinsky D,Quillfeldt JA.Role of hippocampal M1 and M4 muscarinic receptor subtypes in memory consolidation in the rat.Pharmacol Biochem Behav.2003,74(2):411-415.
[79]Ghelardini C,Galeotti N,Matucci R,Bellucci C,Gualtieri F,Capaccioli S,Quattrone A,Bartolini A.Antisense 'knockdowns' of M1 receptors induces transient anterograde amnesia in mice.Ueuropharmacology.1999 Mar;38(3):339-48.
[80]Miyakawa T,WamadaM,DuttaroyA,WessJ.Hyperactivityandintact hippocampus-dependent learning in mice lacking the M1 muscarinic acetylcholine receptor.J Neurosci.2001Jul 15;21(14):5239-50.
[81]黄忠致等.猕猴体感Ⅰ区皮层内注射甲基硫酸新斯的明对痛阈的影响。第十九届中国生理学大会学术论文摘要汇编。生理通讯增刊,1994:112.
[82]徐嵘,关新民,王才源.电针及辣椒素处理对脊髓后角ACHE活性和[3H]-QNB结合位点的影响.针刺研究,1996,21(2):47-51.
[83]关新民,施静,张育文.针刺痛时脊髓后和脊神经节内Ach代谢的变化.针刺研究,1990,15(1):18-23。
[84]关新民,王才源,余福.中枢乙酰胆碱与针刺镇痛关系的研究.针刺研究,1991,16(2):129-137.
[85]魏月娥,傅冠英,万选才.针刺后交感神经中枢内神经介质变化的组织化学观察.解剖学报,1983,14(2):185.
[86]徐维,阎亚生,石宏,陈正秋.乙酸胆碱参与皮层下行调制疼痛的作用.针刺研究,1991,16(3):155-156.
[87]潘玉贞,吴建群,王绍.乙酰胆碱(ACh)兴奋外侧缰核拮抗电针镇痛.长春中医学院学报,1997.13(61).58-59
[88]Lewis JW,Cannon JT,Liebeskind JC.Involvement of central muscarinic cholinergic mechanisms in opioid stress analgesia.Brain Res.1983,270(2):289-93.
[89]Katayama Yet al:Brain Res,296:263.1984.Katayama Y,Watkins LR,Becket DP,Hayes RL.Non-opiate analgesia induced by carbachol microinjection into the pontine parabrachial region of the cat.Brain Res.1984:296(2):263-83.
[90]张立新,刘祚延.微电泳乙酰胆碱和阿托品对大鼠丘脑束旁核痛敏神经元电活动的影响.生理学报,1993,45(1):55-61.
[91]孙明智,范平,张季叶等.乙酰胆碱对丘脑束旁核痛兴奋和痛抑制神经元放电的影响及其机制的探讨,针刺研究,1989,14(1-2):104-105.
[92]徐广银,等.内源性胆碱能系统在佐剂性关节炎大鼠电针镇痛中的作用。第十九届中国生理学大会学术论文摘要汇编,生理通讯增刊,1994,112.
[93]Smith MD,Yang XH,Nha JY,Buccafusco JJ.Antinociceptive effect of spinal cholinergic stimulation:interaction with substance 。 Life Sci.1989;45(14):1255-61。
[94]Hamm RJ,Knisely JS.Developmental differences in the analgesia produced by the central cholinergic system.Dev Psychobiol.1987 May;20(3):345-54.
[1]Megan Long.Acupuncture and medicine:promoting the body' s natural ability to heal itself.(http:/www.michiganacupuncture.org/faq/html)
[2]韩济生.影响针刺镇痛效果的若干因素.针刺研究,1994,19(3-4):1-4.
[3]董志强,马飞,笪翠娣,谢虹,李为民,王彦青,吴根诚.累加电针提高神经痛大鼠背根神经节GDNFmRNA的表达.上海针灸杂志,2005,24(2):33-36.
[4]孙瑞卿,王贺春,景峥,王韵,万有,韩济生.2Hz电针减轻神经源性痛大鼠的痛觉超敏利冷诱发的持续性疼痛.中国疼痛医学杂志,2003,9,(4):220-224.
[5]王贺春,万有,姚磊,韩济生.不同频度电针治疗大鼠慢性神经原性同的疗效比较.针刺研究,2002,27(2):112-118.
[6]王文靖,赵仓焕,谢步霓,盛佑祥,朱小华.不同间隔时间电针对炎症痛大鼠下丘脑脑阿片肽基因表达的影响.针刺研究,2006,31(3):131-135.
[7]罗非.反复电针对慢性痛的累加治疗作用及其机制研究.生理科学进展,1996,7(3):241-244.
[8]陈瑾,刘光谱,唐勇.中枢及外周5-HT、5-HIAA在针刺镇痛后效应中的作用.中医药学刊,2003,21(9):1446-1450.
[9]Bennett GJ,Xie YK.A peripheral mononeuropathy in the rat that produces disorders of pain sensation like those seen in man.Pain,1988,33(1):87-107.
[10]聂发华.电针治疗坐骨神经痛50例.新疆中医,1999,66(2);25.
[11]陈晓红,康妹娟,赵淳,韩济生.大鼠脊髓中三类阿片受体均参与2-15Hz电针镇痛.北京医科大学学报,1992,24(1):54.
[12]刘俊岭,陈淑萍,高永辉,孟凡颖.不同强度、不同频度电针对慢性痛大鼠镇痛作用的比较.针刺研究,2006,31(5):280-285.
[13]王文靖,赵仓焕,谢步霓,盛佑祥,朱小华.不同间隔时间电针对炎症痛大鼠下丘脑脑阿片肽基因表达的影响.针刺研究,2006,31(3):131-135.
[14]Palmon SC,Megan J.Estrogen increases cGMP inselected brain regions and in cerbral microvessels.J Cerb Blood Flow Metab,1998,18(1):1248-1252.
[15]Behl C,Lezoualc' h F.Estrogenswith an intact phenolic group prevent death of neuronal cells following glutathione depletion.Restor Neurol Neurosci,1998,12(223):127-134.
[16]Panikck KS,Guan G,King MA.17beta-estradiol attenuates CREB decline in the rat hippocampus following seizure.J Neurobiol,1997,33(7):961-967.
[17]DianeD,Murphy,NelsonB,et al.Estradiol increasesdendriticspinedensity by reducing GABA neurotransmission in hippocampal neurons.J Neurosci,1998,18(7):2 550-2 559.
[18]Henderson VW.Estrogen,cognition and a woman's risk of Alzheimer's disease [J].Am J Med,1997,103(3A):11S-18S.
[19]Asthana S,Craft S,Baker LD,etal.Cognitive and neuroendocrine response to transdemal estrogen in postmenopauseal woman with Alzheimer's disease:results of placebo controlled,double-blind,pilot study[J].Psychoneuroendocrinology,1999,24(6):657-677.
[201Dong H T,Zhang T S,Chen Y L,etal.Survey and analysis on Alzheimer's disease treated by traditional Chinese medicine[J].Shanghai J Tradit Chin Med(上海中医药杂志),2000,20(4):17-19.
[21]Zhao LX,Chen O,Diaz Brinton,et al.Neuroprotective and neurotrophic efficacy of phytoestrogens in cultured hippocampal neurons[J].Exp Biol Med(Maywood).2002,227(7):509-519.
[22]Dubal DB,Zashon ML,Pettigrew LC,et al.Estradiol protects against ischemic injury[J].J Cereb Blood Flow Metab,1998,18(11):1253-1258.
[23]Birge SJ.Is there a role for estrogen replacement therapy in the prevention and treatment of dementia ? J Am Geriatr 8oc,1996;44(7):865-870.
[24]Singh M,Meyer EM,Simpkins JW.Ovarian steroid deprivation ersults in areversible learning impairment and compromised cholinergic function in female Sprague-Dawley rats.Brain Res,1994;644(2):305-12.
[25]NovelliG,Reichardt JKV.Molecularbasis ofdisorderofhuman galactose m etabolism:past,present,and future[J].Molecular Genetics and Metabolism,2000,71(1):62-65.
[26]Forcier NJ,Mizisin AP,Rmimer M A,et al.Cellular pa-thology of the nerve microenvironment in galactose in-toxication[J].J Neuropathol Exp Neuro,1991,50(3):236-255.
[27]陈勤.抗衰老研究实验方法[M].北京:中国医药科技出版社,1996:494-496.6.
[28]王月华,杜冠华.L-肉毒碱对D-半乳糖衰老模型小鼠的作用.中国老年学杂志, 2003,23(1):49-51.
[29]Hua X,Lei M,Zhang Y,Ding J,tlan Q,Hu G,Xiao M.Long-term D-galactose injection combined with ovariectomy serves as a new rodent model for Alzheimer' s disease.Life Sci.2007;80(20):1897-905.
[30]曲淑君.疼痛的治疗。中国临床康复,2003,6(12):1790-1.
[31]严相默。神经阻滞技术与周围神经痛.中国临床康复,2003,7(6):881-5.
[32]Zheng LF,Wang R,Xu YZ,Yi XN,Zhang JW,Zeng ZC.Calcitonin gene-related peptide dynamics in rat dorsal root ganglia and spinal cord following different sciatic nerve injuries.Brain Res 2008 2;1187:20-32.
[33]宋学军.神经病理性疼痛的动物模型和分子机制.江苏临床医学杂志,2002,6(5):360-399.
[34]赵峰,戴红,郑泓溶,等.中枢性疼痛机制的研究进展.中华物理医学与康复杂志,2003,25(5):310-320.
[35]Bennett GJ,Xie YK.A peripheral mononeuropathy in rat produce disorders of pain sensation like those seen in man.PAIN,1988;33(1):87-107.
[36]Nuytten D,Kupers R,Lamme S,et al.Further evidence for myelinated as well as unmyelinated fiber damage in a rat model of neuropathic pain.Exp Brain Res,1992:91(1):73-8.
[37]Maves TJ,Pechman PS,Gebhart GF,et al.Possible chemical contribution from chromic gut sutures produces disorders of pain sensation like those seen in man.Pain,1993:54(1):57-69.
[38]Attal N,Jazat F,Kayser V,et al.Further evidence for'pain-ralated'behaviour in a model of unilateral peripheral mononeuropathy.Pain,1990;41(2):235-51.
[39]杨向红,王彦青,高秀,吴根诚,电针对大鼠神经痛痛敏分数的影响.针刺研究,2002,27(1):60-63.
[40]董志强,马飞,笪翠娣,谢虹,李为民,王彦青,吴根诚.累加电针提高神经痛大鼠背根神经节GDNFmRNA的表达.上海针灸杂志,2005,24(2):33-36。
[41]李忠仁主编,实验针灸学,中国中医药出版社,2003年1月第一版:174.
[42]Giancar P,Giacomo S.Nootropic drugs and brain cholinergic mechanism[J].Pro Neuro Psychopharmacol and Biol Psychiat,1989,13:77.
[43]孟凡颖,屯针“足三里”-“阳陵泉”穴区治疗慢性痛累积效应机制的分析,2007,硕士论文文库。
[44]刘洁生,谢丽玲,洪岸,等.神经元烟碱乙酰胆碱受体受阻滞所致老年痴呆动物模型的建立.卫生研究,1999,28(4):32-35.
[45]Mesulam MM,Van Hoesen GW.Acetylcholinesterase-rich projections from the basal forebrain of the rhesus monkey to neocortex.Brain Res.1976 Jun 4;109(1):152-7.
[46]张艳,熊希凯.骨骼肌运动终板和神经纤维的酶组织化学及镀银双重显示.中国组织化学与细胞化学杂志,1998,(7):364.
[47]Mesulam M-M,Volicer L,Marquis JK,et al.Systematic regional differentces in the cholinergic innervation of primate cerebral cortex:distribution of enzyme activities and some behavioral implications.Ann Neurol,1986,19:144-151.
[48]Geula C,Mesulam MM.Cortical cholinergic fibers in aging and Alzheimer' s disease:a morphometric study[J].Neuroscience,1989,33:469-481.
[49]李质馨,田洪艳,鲁质博,等.胎脑提取液对坐骨神经损伤大鼠脊髓运动神经元酶活性的影响.解剖学杂志,2003,26(5):443-446.
[50]薛景凤,杨振军,李健.坐骨神经局部缺血后相关神经元胞体乙酰胆碱酯酶变化的定量分析.解剖学杂志,2007,30(1):54-56.
[51]杨军,王静,姜文,等.赤勺总甙对D-半乳糖衰老小鼠学习记忆及代谢产物的影响.中国药理学通讯,2001,17(6):697.
[52]Rodriguez-Puertas R,Pascual J,Vilaro T,et al · Autoradiographic distribution of M1,M2,M3,and M4 muscarinic receptor subtypes in Alzheimer' s disease [J]· Synapse,1997;26(4):341-50 ·
[53]Pauly JR·Nicotinic cholinergic receptor deficits in Alzheimer' s disease:Where' s the smoke[J]J Alzheimers Dis,1999:1(4,5):221-30·
[54]王才源,等.电针镇痛对太鼠脑内ACh,ChAc及AChE活性的影响.针刺研究,1987,12(2):60.
[55]关新民,施静,张育文.针刺痛时脊髓后和脊神经节内Ach代谢的变化.针刺研究,1990,15(1):18-23。
[56]林真.试析乙酰胆碱酯酶活性传变与针灸作用及经络现象的关系.中国针灸,1996,43(1):43-45.
[57]王少锦,李爱英,张雪静.针刺督脉穴对拟AD大鼠海马区Ache、ChAT活性的影响.南京中医药大学学报,2004,20(5):289-291。
[58]柏志全,郑辉.姚平.电针对大鼠脑缺血再灌注后脑组织内乙酰胆碱酯酶活性的影响.中国临床康复2004,5(4):672-673.
[59]Naguib M,Yaksh TL.Characterization of muscarinic receptor subtypes that mediate antinociception in the rat spinal cord.。 Anesth Analg.1997:85(4):847-53.
[60]Shiokawa M,Yamaguchi T,Narita M,Okutsu D,Nagumo Y,Miyoshi K,Suzuki M,Inoue T,Suzuki T.Effects of fasudil on neuropathic pain-like state in mice。 Nihon Shinkei Seishin Yakurigaku Zasshi.2007;27(4):153-9.
[61]Malmberg AB,Brandon EP,Idzerda RL,Liu H,McKnight GS,Basbaum AI.Diminished inflammation and nociceptive pain with preservation of neuropathic pain in mice with a targeted mutation of the type Ⅰ regulatory subunit of cAMP-dependent protein kinase.J Neurosci.1997,17(19):7462-70.
[62]Yajima Y,Narita M,Shimamura M,Narita M,Kubota C,Suzuki T.Differential involvement of spinal protein kinase C and protein kinase A in neuropathic and inflammatory pain in mice.Brain Res.2003,992(2):288-93.
[63]Miletic G,Pankratz MT,Miletic V.Increases in the phosphorylation of cyclic AMP response element binding protein(CREB) and decreases in the content of calcineurin accompany thermal hyperalgesia following chronic constriction injury in rats.Pain.2002,99(3):493-500.
[64]王哲,随瑜,德亮,炜熙.性应激对大鼠海马神经元PKA和P-CREB蛋白表达的影响及抗抑郁剂的拮抗作用.中南大学学报,2006,31(5):767-771.
[65]洪小平,刘能保,张敏海,等.慢性复合应激对大鼠学习和记忆功能及海马内PKA-C _β表达的影响.华中科技大学学报,2005,34(3):257-261.
[66]Liang Z,Liu F,6rundke-Iqbal I,Iqbal K,Gong CX.Down-reguiation of cAMP-dependent protein kinase by over-activated calpain in Alzheimer disease brain.J Neurochem.2007,103(6):2462-70.
[67]Tanaka Y,Takase M,6amo S.Relationship between general anesthesia and memory in Drosophila involving the cAMP/PKA pathways and adhesion-related molecules.Curr Med Chem.2007;14(13):1479-88.
[68]Takeo S,Niimura M,Miyake-Takagi K,et al.A possible mechanism for improvement by a cognition-enhancer nefiracetam of spatial memory function and c AMP - mediated signal transduction system in sustained cerebral ischaemia in rats.Br J Pharmacol 2003;138(4):642-54.
[69]Bernabeu R,Bevilaqua L,Ardenghi,et al.Involvement of hippocapal c AMP/c AMP-dependent protein kinase signaling pathways in a late memory consolidation phase of aversively motivated learning in rats.Proc Natl Acad Sci U S A 1997,94(13):7041-6.
[70]Waltereit R,Weller M.Signaling from c AMP/PKA to MAPK and synaptic plasticity.Mol Neurobiol 2003,27(1):99-106.
[71]Nagakura A,Niimura M,Takeo S.Effects of a phosphodiesterase Ⅳ inhibitor rolipram on microsphere embolism-induced defects in memory function and cerebral cyclic AMP signal transduction system in rats.Br J Pharmacol 2002,135(7):1783-93.
[72]常铉,舒斯云,包新民,江刚.PKA-CREB信号通路在大鼠纹状体边缘区及海马空间学习记忆过程中的作用.中医临床康复,2005,9(1):39-41.
[73]Smith JA,Davis CL,Burgess GM.Prostaglandin E2--induced sensitization of bradykinin--evoked responses in rat dorsal root ganglion neurons is medlated by cAMP --dependent protein kinase A.Eur J Neuorsci.2000:12(9):3250-3258.
[74]Malmber AB,Brandon EP,Idzerda RL,et al.Diminished inflammation and nociceptive pain with preservation of neuropathic pain in mice with a targeted mutation of the type Ⅰ regulatory subunit of c AMP-dependent protein kinase.J Neurosci 1997;17(19):7462-70.
[751Codere TJ.YashPal K.Intracellular messengers contributing to Persistent nociception and hyperalgesia induced by-Lglutamate and substance P in the rat formalin pain model.Eur J Neuorsci.1994;6(8):1328--1334.
[76]姚永兴,张励才,宋学军,曾冈明.鞘内注射H-89对神经病理痛大鼠脊髓背角磷酸化cAMP 反应元件结合蛋白表达的影响.中华麻醉学杂志,2005,25(5):371-374.
[77]Shiokawa M,Yamaguchi T,Narita M,Okutsu D,Nagumo Y,Miyoshi K,Suzuki M,Inoue T,Suzuki TEffects of fasudil on neuropathic pain-like state in mice.Nihon Shinkei Seishin Yakurigaku Zasshi.2007 Aug;27(4):153-9.
[78]陆汎、朱红梅、谢俊杰,周海虹,陈艳玲,胡纪原。屯针对抑郁症模型大鼠行为学、血浆COR及海马PKA、PKC表达的影响。中国针灸,2008,28(3):214-8.
[79]张敏,图娅.电针对慢性应激抑郁模型大鼠学习记忆行为的影响.中医研究,2008,21(3):15-18。
[80]付弋,梁繁荣,陶巧林.不同针法对佐刑性关节炎大鼠下丘脑镇痛效应比较.上海针灸杂志,2005,24(5):38-41.