捻转补泻手法对应激性高血压大鼠Ach-NO-cGMP信号转导通路影响的机制研究

【作者】 刘炜；

【导师】 刘清国；

【作者基本信息】 北京中医药大学 ， 针灸推拿学， 2011， 硕士

【摘要】 目的观察针刺捻转补泻手法对应激性高血压大鼠模型Ach-NO-cGMP信号转导通路中乙酰胆碱(Ach)、一氧化氮合成酶(NOS)、环鸟苷酸(cGMP)表达的影响,揭示捻转补泻手法与效应的相关性及捻转补泻手法产生不同生物学效应的机制,以期为针刺捻转补泻手法的实用性和科学性提供依据,促进针刺补泻手法在针灸临床中的应用和发展。方法清洁级健康雄性Wistar大鼠60只,体重220～280g。按照随机数字表分为5组,每组12只,A：正常组、B：模型对照组、C：针刺对照组、D：针刺补法组、E：针刺泻法组五组。空白组不做处理,其他各组均采用电击加噪声刺激法制成应激性高血压动物模型,并在造模的前1天,造模第3、6、9、12、15天的刺激后2小时测量大鼠收缩压。15天造模成功后,每天上午分别对针刺组、补法组、泻法组大鼠的双侧“太冲”穴施以针刺不做手法、捻转补法和捻转泻法操作。捻转手法幅度为360°/次,60次/min,持续捻针1min,留针20min。治疗7天后,对实验动物进行断头处死,取血和胸主动脉,用ELISA法检测血浆和胸主动脉组织中的Ach、NOS、cGMP含量。结果1应激性刺激和不同针刺手法对各组大鼠收缩压的影响经过15天应激性造模后,各造模组大鼠收缩压显著上升,与空白组相比有显著差异(p<0.01),并且造模各组之间没有显著差异(p>0.05)；经过7天针刺处理之后,B组大鼠血压波动不明显,与A组比较具有显著差异(p<0.01)；C组、D组、E组血压均下降,E组血压下降最显著,D组和针刺组与E组比较差异显著(p<0.05)；D组、E组与B组比较,血压显著下降(p<0.01)；C组与B组比较,血压下降(p<0.05)。2针刺捻转补泻对各组大鼠血浆中Ach的影响B组大鼠血浆中Ach显著下降,与A组相比有显著差异(p<0.01)；C组、D组、E组大鼠血浆中Ach含量均上升,与B组比较具有显著差异(p<0.01)；E组大鼠血浆中Ach上升最明显,并与c组和D组比较均有显著差异(p<0.01),C组和D组大鼠血浆中Ach上升,二者比较差异不显著(p>0.05)。3针刺捻转补泻对各组大鼠主动脉组织中NOS的影响在主动脉组织NOS中,B组主动脉组织NOS含量显著下降,与A组比较差异显著(p<0.01)；C组、D组、E组中NOS含量均上升,C组NOS含量上升与B组比较有差异(p<0.05)；D组和E组上升显著,并与B组比较差异显著(p<0.01)；其中D组上升最显著,D组与C组比较有差异(p<0.05)；E组和C组比较NOS含量无明显差异(p>0.05),D组和E组NOS含量上升没有明显差异(p>0.05)。4针刺捻转补泻对各组大鼠主动脉组织中cGMP的影响在主动脉cGMP中,B组主动脉组织cGMP含量显著下降,与A组比较差异显著(p<0.01)；C组、D组、E组中cGMP含量与造模组比较均上升,以D组上升最明显,D组、E组与B组比较cGMP含量上升具有显著差异(p<0.01)；C组与E组比较cGMP含量上升具有差异(p<0.05)；C组与D组比较cGMP含量上升具有显著差异(p<0.01),D组和E组比较,cGMP含量上升不具有显著差异(p>0.05)。结论1捻转补泻手法有降低应激性高血压的作用,捻转补法和捻转泻法的降压效应存在差异；2捻转补泻手法能升高应激性高血压大鼠模型中血浆Ach、主动脉组织NOS、主动脉组织cGMP的含量；3捻转补泻手法调节血压的机制之一可能是通过Ach-NO-cGMP信号转导通路实现的。更多还原

【Abstract】 ObjectiveThe research aims at observing the effect on Ach, NOS, cGMP on Ach-NO-cGMP singal transduction pathway of stress-induced hypertension rats treated by twirling reinforcing and reducing manipulation, revealing the mechanism of relativity between twirling reinforcing and reducing manipulation and the different biology effect, providing an experimental evidence to twirling reinforcing and reducing manipulation, and promoting the popularization of manipulation in acupuncture practice.Method60 clean male Wistar rats with weight ranging from 220g to 280g were divided into A(blank group), B(model group), C(needling group), D(reinforcing group), and E(reducing group) five groups randomly. Electric shock and noise were applied to stimulate the rats into stress-induced hypertension models except group A. Each rat was treated by 1 minute manipulation and 20 minutes needle retaining. The treatment lasted for 7 days and once each day. After the treatment Ach, NOS, cGMP contents in plasma and aortic were detected by means of radio immuno assay and ELISA.Result1 Systolic blood pressure changes of stress-induced hypertension rats after electric shock and noise stimulation and acupuncture treatmentThe systolic blood pressure of each stimulated group increased significantly after 15 days lectric shock and noise stimulation. Campared with group A the difference were significant (p<0.01). The differences of each stimulated group were not obvious (p>0.05). After 7 treatment days, the systolic blood pressure of group B were stable and different to the group A (p<0.01); the the systolic blood pressure of group C, group D and group E all decreased, the group E were the most obvious, and the group D and group C were sinificient different to group E (p<0.05); group D and group E campared with group B were obviously different (p<0.01); and the differences of systolic blood pressure were significant between group C and group B (p<0.05).2 Changes of Ach in plasma of stress-induced hypertension ratsThe Ach in plasma decreased significantly in group B, which were obviously different to group A (p<0.01); the Ach of group C,D and E all increased and different to group B (p<0.01); the Ach of group E inceased most obviously, which were different to group C and group D (p<0.01); however, there were no signifcient difference in group C and D (p>0.05). 3 Changes of NOS in aortic of stress-induced hypertension ratsThe NOS in aortic decreased significantly in group B, which were obviously different to group A (p<0.01); the NOS of group C,D and E all increased, there were significient differences between group C and group B (p<0.05); campared group D and E with group B, the differences were obvious (p<0.05), in which the NOS in group D was the highest; there were different between group D and group C (p<0.05); however, there were no signifient differences between group E and C and group D and E (p>0.05).3 Changes of cGMP in aortic of stress-induced hypertension ratsThe cGMP in plasma decreased significantly in group B, which were obviously different to group A (p<0.01); the cGMP of group C,D and E all increased, in which the group D was the highest; campared group D and E with group B, the differences were obvious (p<0.01) campared group C and group E the differences were signifcient (p<0.05); campared group C and group D the differences were more signifcient (p<0.01); however, there were no signifient differences between group E and C and group D and E (p>0.05)Conclusion1 Twirling reinforcing and reducing manipulation could reduce stress-induced hypertension, the antihypertensive effect are difference between twirling reinforcing and reducing manipulation;2 Twirling reinforcing and reducing manipulation could increase the contents of Ach, NOS, cGMP in plasma and aortic of stress-induced hypertension rats;3 Ach-NO-cGMP singal transduction pathway could one of the blood pressure regulating mechanism of twirling reinforcing and reducing manipulation.更多还原