电针“曲池”穴对中枢血管紧张素Ⅱ诱发的动脉血压升高大鼠异常交感神经活性的影响
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摘要
目的:观察电针"曲池"穴对侧脑室慢性灌流血管紧张素Ⅱ(AngⅡ)高血压大鼠平均动脉压、交感神经活性、压力感受敏感性,以及延髓头端腹外侧区(RVML)内脑源性神经生长因子(BDNF)和还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)酶亚单位p 47phox表达的影响。方法:SD大鼠随机分为假手术组、aCSF组、模型组、曲池组、肩髃组,每组9只。其中模型组、曲池组、肩髃组大鼠使用Alzet微渗透泵分别经侧脑室慢性灌流AngⅡ(200μg·kg-1·d-1),而aCSF组给予人工脑脊液(aCSF)灌流。曲池组给予电针"曲池"穴治疗,"肩髃"组给予电针"肩髃"穴治疗,均每日1次,每次20min,共治疗2周。使用无创系统测量清醒大鼠的平均动脉压(MAP)、心率(HR)、24h尿去甲肾上腺素(NE)水平和压力感受敏感性;Real-time PCR和Western blot方法检测RVML内BDNF和p 47phox mRNA和蛋白的表达。结果:与aCSF组比较,模型组大鼠的MAP、HR和24h尿NE水平均显著增加(P<0.01),压力反射敏感性显著降低(P<0.01)。与模型组比较,曲池组大鼠的MAP、HR和24h尿NE水平显著降低(P<0.05),压力反射敏感性显著增加(P<0.05)。曲池组的RVML内BDNF mRNA和蛋白水平均较模型组显著升高(P<0.05);而曲池组RVML内p 47phox mRNA和蛋白的表达与模型组比较显著下降(P<0.05)。结论:电针"曲池"穴可抑制中枢AngⅡ诱发的动脉血压升高和交感神经活性增加,并提高压力感受敏感性。该效应可能是通过上调RVML内BDNF,并抑制脑内病理性血管紧张素Ⅱ引发的异常交感神经活性而达到的。
Objective To observe the effect of electroacupuncture(EA)stimulation of"Quchi"(LI 11)on arterial blood pressure(BP),sympathetic nerve activity,barorefiex sensitivity(BRS)and expression of brain derived neurotrophic factor(BDNF)and nicotinamide adenine dinucleotide phosphate(NADPH)enzyme subunit p 47 phox in the rostral ventrolateral medulla(RVML)in hypertension rats.Methods A total of 45 male SD rats were randomly divided into sham operation,artificial cerebro-spinal fluid(aCSF),model,"Quchi"(LI 11),and Jianyu(LI 15)groups(n=9in each group).The hypertension model was established by microinjection of AngⅡ(200μg·kg-1·day-1)into the lateral ventricle(A-C:1.0mm,L:1.4mm,H:4.5mm)for 2weeks(aCSF for aCSF group).After 1week's AngⅡ perfusion,EA(2Hz/15 Hz,1mA)was applied to bilateral"Quchi"(LI 11)or"Jianyu"(LI 15)for 20 min,once daily for two weeks.The mean arterial pressure(MAP)and heart rate(HR)were detected by using a Non-Invasive Blood Pressure System(tail cuff method).The BRS(BP-to-HR transfer function)was determined by calculating the ratio of HR fluctuation(HR∶HR,between post-and pre-intravenous injection of phenylephrine)/MAP fluctuation(MAP∶MAP,between post-and pre-phenylephrine injection),the urinary norepinephrine(NE)level in 24 hwas assayed by ELISA.Real-time PCR and Western blot were used to detect the mRNA and protein expression of BDNF and p 47 phox in the RVML region tissue.Results Following modeling,the MAP,HR and 24h-urinary NE levels and p 47 phox mRNA and protein expression levels in the RVML were significantly increased(P<0.01),and the BRS was decreased significantly(P<0.01).After EA intervention,the MAP,HR and 24h-urinary NE levels and p 47 phox mRNA and protein expression levels in the RVML were considerably lower in the LI 11 group than in the model group(P<0.05),while the BRS level was markedly increased in the LI 11 group compared with the model group(P<0.05).The expression levels of BDNF mRNA and protein in the RVLM region were obviously up-regulated in the LI 11 group compared with the model group(P<0.05).No significant differences were found between the LI 15 group and the model group in the MAP,HR,24h-urinary NE,BRS,BDNF mRNA and protein and p 47 phox mRNA and protein expression levels(P>0.05).Conclusion EA stimulation of"Quchi"(LI 11)can down-regulate arterial blood pressure,sympathetic nerve activity,and increase the baroreflex sensitivity in hypertension rats,which may be related to its effects in down-regulating p 47 phox mRNA and protein expression in the RVML.
Objective To observe the effect of electroacupuncture(EA)stimulation of"Quchi"(LI 11)on arterial blood pressure(BP),sympathetic nerve activity,barorefiex sensitivity(BRS)and expression of brain derived neurotrophic factor(BDNF)and nicotinamide adenine dinucleotide phosphate(NADPH)enzyme subunit p 47 phox in the rostral ventrolateral medulla(RVML)in hypertension rats.Methods A total of 45 male SD rats were randomly divided into sham operation,artificial cerebro-spinal fluid(aCSF),model,"Quchi"(LI 11),and Jianyu(LI 15)groups(n=9in each group).The hypertension model was established by microinjection of AngⅡ(200μg·kg-1·day-1)into the lateral ventricle(A-C:1.0mm,L:1.4mm,H:4.5mm)for 2weeks(aCSF for aCSF group).After 1week's AngⅡ perfusion,EA(2Hz/15 Hz,1mA)was applied to bilateral"Quchi"(LI 11)or"Jianyu"(LI 15)for 20 min,once daily for two weeks.The mean arterial pressure(MAP)and heart rate(HR)were detected by using a Non-Invasive Blood Pressure System(tail cuff method).The BRS(BP-to-HR transfer function)was determined by calculating the ratio of HR fluctuation(HR∶HR,between post-and pre-intravenous injection of phenylephrine)/MAP fluctuation(MAP∶MAP,between post-and pre-phenylephrine injection),the urinary norepinephrine(NE)level in 24 hwas assayed by ELISA.Real-time PCR and Western blot were used to detect the mRNA and protein expression of BDNF and p 47 phox in the RVML region tissue.Results Following modeling,the MAP,HR and 24h-urinary NE levels and p 47 phox mRNA and protein expression levels in the RVML were significantly increased(P<0.01),and the BRS was decreased significantly(P<0.01).After EA intervention,the MAP,HR and 24h-urinary NE levels and p 47 phox mRNA and protein expression levels in the RVML were considerably lower in the LI 11 group than in the model group(P<0.05),while the BRS level was markedly increased in the LI 11 group compared with the model group(P<0.05).The expression levels of BDNF mRNA and protein in the RVLM region were obviously up-regulated in the LI 11 group compared with the model group(P<0.05).No significant differences were found between the LI 15 group and the model group in the MAP,HR,24h-urinary NE,BRS,BDNF mRNA and protein and p 47 phox mRNA and protein expression levels(P>0.05).Conclusion EA stimulation of"Quchi"(LI 11)can down-regulate arterial blood pressure,sympathetic nerve activity,and increase the baroreflex sensitivity in hypertension rats,which may be related to its effects in down-regulating p 47 phox mRNA and protein expression in the RVML.
引文
[1]张琪,谭颖颖.针刺降压的交感神经机制研究进展[J].陕西中医学院学报,2012,35(5):98-101.
[2]杨佃会,周亮,贾真,等.曲池穴治疗高血压病探讨[J].山东中医药大学学报,2004,28(5):338-339.
[3]Gabor A,Leenen F H.Central neuromodulatory pathways regulating sympathetic activity in hypertension[J].J Appl Physiol,2012,113(8):1294-1303.
[4]Arnold A C,Gallagher P E,Diz D I.Brain renin-angiotensin system in the nexus of hypertension and aging[J].Hypertens Res,2013,36(1):5-13.
[5]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[J].中国针灸,2009,29(5):349-352.
[6]邹艳,吕小笑,罗志平.曲池穴捻转补泻法治疗原发性高血压疗效观察[J].上海针灸杂志,2015,34(9):847-849.
[7]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[J].中国中西医结合杂志,2002,22(12):1080-1083.
[8]刘婉宁,金侣位,韩东伟,等.电针“曲池”“足三里”“神门”对自发性高血压大鼠血压及血管紧张素Ⅱ、醛固酮及心钠素的影响[J].针刺研究,2009,34(6):393-398.
[9]马朝阳,王艳富,万文俊,等.电针曲池对原发性高血压患者血浆NPY与NT浓度的影响[J].新中医,2011,43(4):89-91.
[10]钱春艳,陈邦国,谢霞,等.电针曲池穴对二肾一夹高血压大鼠血压和颈总动脉血管重构的影响[J].陕西中医,2008,29(8):1079-1080.
[11]宿中笑,肖达,雎久红,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[J].上海针灸杂志,2003,22(5):20-22.
[12]Zhang Q,Yao F,Raizada M K,et al.Apelin gene transfer into the rostral ventrolateral medulla induces chronic blood pressure elevation in normotensive rats[J].Circ Res,2009,104(12):1421-1428.
[13]张琪,谭颖颖.脑内活性氧抑制对醋酸去氧皮质酮-盐型高血压大鼠交感神经活动的影响[J].南方医科大学学报,2014,34(11):1632-1636.
[14]Lob H E,Schultz D,Marvar P J,et al.Role of the NADPH oxidases in the subfornical organ in angiotensinⅡ-induced hypertension[J].Hypertension,2013,61(2):382-387.
[15]Chan S H,Chan J Y.Angiotensin-generated reactive oxygen species in brain and pathogenesis of cardiovascular diseases[J].Antioxid Redox Signal.2013,19(10):1074-1084.
[16]Zhang Q,Yao F,O’Rourke S T,et al.AngiotensinⅡenhances GABA(B)receptor-mediated responses and expression in nucleus tractus solitarⅡof rats[J].Am J Physiol Heart Circ Physiol.2009,297(5):H 1837-1844.
[17]张琪,谭颖颖.侧脑室慢性灌流(pyr 1)apelin-13对大鼠心血管功能的影响[J].中国药理学通报,2014,30(1):144-145.
[18]林文注,王佩.实验针灸学[M].上海:上海科学技术出版社,1999:123-124.
[19]张琪,谭颖颖.血管紧张素1-7减弱醋酸去氧皮质酮盐型高血压大鼠的左心室重构[J].中国药理学通报,2014,30(7):994-999.
[20]Bali A,Jaggi A S.Angiotensin as stress mediator:role of its receptor and interrelationships among other stress mediators and receptors[J].Pharmacol Res,2013,76:49-57.
[21]Marc Y,Llorens-Cortes C.The role of the brain renin-angiotensin system in hypertension:implications for new treatment[J].Prog Neurobiol,2011,95(2):89-103.
[22]Paton J F,Wang S,Polson J W,et al.Signalling across the blood brain barrier by angiotensinⅡ:novel implications for neurogenic hypertension[J].J Mol Med(Berl),2008,86(6):705-710.
[23]Chan S H,Wu C W,Chang A Y,et al.Transcriptional upregulation of brain-derived neurotrophic factor in rostral ventrolateral medulla by angiotensinⅡ:significance in superoxide homeostasis and neural regulation of arterial pressure[J].Circ Res,2010,107(9):1127-1139.
[2]杨佃会,周亮,贾真,等.曲池穴治疗高血压病探讨[J].山东中医药大学学报,2004,28(5):338-339.
[3]Gabor A,Leenen F H.Central neuromodulatory pathways regulating sympathetic activity in hypertension[J].J Appl Physiol,2012,113(8):1294-1303.
[4]Arnold A C,Gallagher P E,Diz D I.Brain renin-angiotensin system in the nexus of hypertension and aging[J].Hypertens Res,2013,36(1):5-13.
[5]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[J].中国针灸,2009,29(5):349-352.
[6]邹艳,吕小笑,罗志平.曲池穴捻转补泻法治疗原发性高血压疗效观察[J].上海针灸杂志,2015,34(9):847-849.
[7]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[J].中国中西医结合杂志,2002,22(12):1080-1083.
[8]刘婉宁,金侣位,韩东伟,等.电针“曲池”“足三里”“神门”对自发性高血压大鼠血压及血管紧张素Ⅱ、醛固酮及心钠素的影响[J].针刺研究,2009,34(6):393-398.
[9]马朝阳,王艳富,万文俊,等.电针曲池对原发性高血压患者血浆NPY与NT浓度的影响[J].新中医,2011,43(4):89-91.
[10]钱春艳,陈邦国,谢霞,等.电针曲池穴对二肾一夹高血压大鼠血压和颈总动脉血管重构的影响[J].陕西中医,2008,29(8):1079-1080.
[11]宿中笑,肖达,雎久红,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[J].上海针灸杂志,2003,22(5):20-22.
[12]Zhang Q,Yao F,Raizada M K,et al.Apelin gene transfer into the rostral ventrolateral medulla induces chronic blood pressure elevation in normotensive rats[J].Circ Res,2009,104(12):1421-1428.
[13]张琪,谭颖颖.脑内活性氧抑制对醋酸去氧皮质酮-盐型高血压大鼠交感神经活动的影响[J].南方医科大学学报,2014,34(11):1632-1636.
[14]Lob H E,Schultz D,Marvar P J,et al.Role of the NADPH oxidases in the subfornical organ in angiotensinⅡ-induced hypertension[J].Hypertension,2013,61(2):382-387.
[15]Chan S H,Chan J Y.Angiotensin-generated reactive oxygen species in brain and pathogenesis of cardiovascular diseases[J].Antioxid Redox Signal.2013,19(10):1074-1084.
[16]Zhang Q,Yao F,O’Rourke S T,et al.AngiotensinⅡenhances GABA(B)receptor-mediated responses and expression in nucleus tractus solitarⅡof rats[J].Am J Physiol Heart Circ Physiol.2009,297(5):H 1837-1844.
[17]张琪,谭颖颖.侧脑室慢性灌流(pyr 1)apelin-13对大鼠心血管功能的影响[J].中国药理学通报,2014,30(1):144-145.
[18]林文注,王佩.实验针灸学[M].上海:上海科学技术出版社,1999:123-124.
[19]张琪,谭颖颖.血管紧张素1-7减弱醋酸去氧皮质酮盐型高血压大鼠的左心室重构[J].中国药理学通报,2014,30(7):994-999.
[20]Bali A,Jaggi A S.Angiotensin as stress mediator:role of its receptor and interrelationships among other stress mediators and receptors[J].Pharmacol Res,2013,76:49-57.
[21]Marc Y,Llorens-Cortes C.The role of the brain renin-angiotensin system in hypertension:implications for new treatment[J].Prog Neurobiol,2011,95(2):89-103.
[22]Paton J F,Wang S,Polson J W,et al.Signalling across the blood brain barrier by angiotensinⅡ:novel implications for neurogenic hypertension[J].J Mol Med(Berl),2008,86(6):705-710.
[23]Chan S H,Wu C W,Chang A Y,et al.Transcriptional upregulation of brain-derived neurotrophic factor in rostral ventrolateral medulla by angiotensinⅡ:significance in superoxide homeostasis and neural regulation of arterial pressure[J].Circ Res,2010,107(9):1127-1139.