电针人中穴对脑梗死大鼠血管紧张素Ⅱ及其受体基因及蛋白表达的影响

Effect of Acupuncture at Renzhong(DU26) on AngiotensinⅡ Receptor Gene and Protein Expression in Rats with Cerebral Infarction

目的：本研究以MCAO大鼠为研究对象,通过观察脑血流、Ang II及受体的基因和蛋白表达的变化以探讨电针对脑梗死大鼠血管舒缩的干预机制。方法：将90只Wister大鼠随机分为空白组、模型组、电针组,模型组和电针组按术后3、6、12、24 h,3、7、12 d各分为7个时相组,每时相各6只。各组大鼠进行脑血流、实时荧光定量PCR及免疫蛋白印迹的检测。结果：脑血流检测显示模型组与电针组趋势相同,但电针组在1 h、2 h、3 h、12 h显著高于模型组（P〈0.01）。AGT基因表达在梗死后呈增长的趋势,至9 h后显著高于空白组（P〈0.05,P〈0.01）,电针组均低于模型组,且在12 h、15 h有显著性差异（P〈0.05）,至18 h后明显高于空白组（P〈0.01）。脑梗死后AngⅡ蛋白的表达显著增加（P〈0.05,P〈0.01）,但是针刺在梗死后3 h以内及18 h和24 h可显著降低其表达（P〈0.05,P〈0.01）。AT1R基因表达在梗死后3 h内显著增加（P〈0.01）,此后在6 h降至谷底后再次明显增加（P〈0.05,P〈0.01）,电针组表达均低于模型组,且在梗死后3 h内、15 h到24 h时相均有显著性差异（P〈0.05,P〈0.01）,AT1R蛋白表达在造模后显著增加（P〈0.01）,而针刺可下调其表达,除6 h、9 h时相外均有显著性差异（P〈0.05,P〈0.01）。AT2R基因自脑梗死后2 h显著增加（P〈0.01）,在脑梗死后18 h内针刺显著上调了其表达（P〈0.05,P〈0.01）,模型组AT2R蛋白在脑梗死后呈上升趋势,自9h后显著高于空白组（P〈0.05）,电针可提高各时相AT2R的表达,且在1 h、2 h、9 h、21 h、24 h有显著性差异（P〈0.05,P〈0.01）。结论：电针对MCAO大鼠Ang II及受体的基因及蛋白表达有良性调整作用,可促进缺血半暗带的血管舒张。

Objective： In this study,MCAO rats were as the research object. By observing the cerebral blood flow,changes in gene and protein expression of Ang II receptors were observed to explore the intervention mechanism for electrical cerebral vasomotor rats. Methods： Ninety Wister rats were randomly divided into three groups,namely blank group,model group,EA group. The model group was divided into phase groups： 3,6,12,24 hours after operation and EA group into 3,7 and 12 days after operation,6 in each phase group. Each group tested brain blood flow and carried out real-time PCR and Western blot. Results： Cerebral blood flow measurement showed that the model group＇s was the same as that of the EA group. But at 1 h,2 h,3 h and 12 h,EA groups＇ cerebral blood flow was significantly higher than those in the model group（P〈 0. 01）. AGT gene expression had a growing trend which was significantly higher than that of the blank group 9 h later（P〈 0. 05,P〈 0. 01）. EA group each phase group＇s was lower than that of the model group,and at 12 h and 15 h there were significant differences（P〈 0. 05）. At 18 h,it was significantly higher than that of the blank group（P〈 0. 01）. After cerebral infarction,the expression of AngⅡ protein was significantly increased（P〈 0. 05,P〈 0. 01）,but acupuncture after infarction within 3 h,18 h and 24 h can significantly reduce the expression（P〈 0. 05,P〈 0. 01）. AT1 R gene expression was significantly increased after infarction within 3 h（P〈 0. 01）,dropped to the bottom at 6 h and then again increased significantly（P〈 0. 05,P〈 0. 01）. The expression of EA group was lower than that of the model group at each time,and within 3 h,15 h and 24 h after infarction,there were significant differences（P〈 0. 05,P 0. 01）. After modeling,AT1 R protein was significantly increased（P〈 0. 01） and acupuncture can lower its expression,in addition to 6 h and 9 h phase,there were significant differences（P〈 0. 05,P〈 0. 01）. There was a significant increase in AT2 R gene with 2 h after cerebral infarction after（P〈 0. 01） and acupuncture significantly raised its expression within 18 h after cerebral infarction（P〈 0. 05,P〈 0. 01）. The model group＇s AT2 R protein after cerebral infarction was rising,and significantly higher than that of the blank group after 9 h（P〈 0. 05）,EA can improve each phase＇s AT2 R expressions,and at 1 h,2 h,9 h,21 h and 24 h there was a significant difference（P〈 0. 05,P〈 0. 01）. Conclusion： EA has a healthy adjustment for acute cerebral ischemia rats＇ Ang II receptor gene and protein expression and can promote ischemic penumbra vasodilation.