电针对心肌缺血大鼠细胞凋亡指数及miRNAs表达影响的研究

Effect of Electroacupuncture on mi RNAs Expression and Apoptosis in Myocardial Ischemia Rats with Myocardial Ischemia

目的:探讨电针内关穴(电针组)对心肌缺血大鼠细胞凋亡相关微小核糖核酸(miRNA)表达的影响,以期探讨其作用机制。方法:30只Wistar雄性大鼠随机分为正常组、模型组和电针组,每组10只。模型组和电针组以盐酸异丙肾上腺素2 mg/(kg·d)背部皮下注射14 d的方法制备心肌缺血大鼠模型;正常组注射等量生理盐水。电针组在造模后选取内关穴进行电针,每天1次,共21 d。运用酶联免疫吸附试验(ELISA)检测各组大鼠血清CK、CK-MB及VCAM-1的表达量;TUNEL法检测心肌细胞凋亡指数(AI)值;Real time PCR法检测心肌细胞miRNA-133a、miRNA-133b、miRNA-208、miRNA-499的表达量。结果:与正常组比较,模型组大鼠血清CK、CK-MB、VCAM-1表达升高;心肌细胞凋亡指数增多;心肌细胞miRNA-133a、miRNA-133b表达降低,miRNA-208、miRNA-499表达升高,差异均有统计学意义(P<0.01)。电针组上述指标均明显低于模型组,差异有统计学意义(P<0.01)。结论:电针内关穴可有效防治心肌缺血,其保护作用可能与上调心肌miRNA-133a、miRNA-133b的表达,抑制miRNA-208、miRNA-499的表达有关。

Objective： To investigate the effect of electroacupuncture（ EA） at Neiguan（ PC6） on Micro RNA（ miRNA）expression and apoptosis in rats with myocardial ischemia so as to explore its mechanism. Methods： Thirty female Wistar rats were randomly divided into three groups,i. e.,the normal group,the model group and the EA group,10 in each group. Isoprenaline hydrochloride（ ISO） injection with the daily dose 2 mg / kg was subcutaneously injected for 14 days to establish the myocardial ischemia model. Rats in normal group were subcutaneously injected with the equal volume of normal saline. EA at Neiguan（ PC6） was applied for rats in the EA group while modeling,once a day,for 21 successive days. The contents of CK,CK- MB and VCAM- 1 in serum were detected by enzyme- linked immunosorbent assay（ ELISA）. Apoptosis index（ AI） of myocardial cells was detected by TUNEL method. The expressions of miRNA-133 b,miRNA- 208,miRNA- 133 a and miRNA- 499 were detected by Real time PCR method. Results： Compared with normal group,the serum levels of CK,CK- MB and VCAM- 1 were significantly higher in model group and the apoptosis index significantly increased. The expressions of miRNA- 133 a and miRNA- 133 b were decreased but miRNA- 208 and miRNA- 499 significantly increased and the difference was statistically significant（ P〈0. 01）. The above indexes were significantly lower in the EA group than those in the model group with statistical difference（ P〈0. 01）. Conclusions： EA at Neiguan（ PC6） could prevent and treat myocardial ischemia. The protective effect is possibly correlated with up- regulating the expressions of miRNA- 133 a and miRNA- 133 b and inhibiting its expressions of miRNA- 499 and miRNA- 208.