标本配穴对心肌缺血大鼠心肌沉默信息调节因子1的影响

Effects of manifestation points and root cause points combination on SIRT1 in myocardial ischemia rat model

目的:以心肌细胞沉默信息调节因子1(SIRT1)为切入点,探讨标本配穴防治心肌缺血(MI)的作用机制。方法:将50只SPF级Wistar雄性大鼠随机分为正常组、模型组、防治组(标本配穴防治组)、白藜芦醇组、尼克酰胺组,每组10只。以盐酸异丙肾上腺素制作MI模型;防治组在造模前取双内关、双足三里、关元穴接HANS-200型电针仪预处理10min,疏密波,2^(-1)00Hz,1m A;白藜芦醇组和尼克酰胺组分别在造模前灌胃给予白藜芦醇溶液和尼克酰胺溶液。结束后观察各组心电图S-T段和肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)活性的变化,采用Western blot法检测大鼠心肌细胞中SIRT1蛋白表达的水平。结果:与正常组比较,模型组心电图S-T段和血清CK-MB、LDH值极显著升高(P<0.01),心肌SIRT1蛋白表达极显著下降(P<0.01),提示SIRT1蛋白表达的减少与MI的发生密切相关。与模型组比较,防治组和白藜芦醇组心电图S-T段和血清CK-MB、LDH值均明显下降(P<0.05),心肌SIRT1蛋白表达显著上升,提示标本配穴电针和白藜芦醇均能有效防治心肌缺血损伤,并能有效抑制SIRT1表达的下降;尼克酰胺组心电图S-T段和血清CK-MB、LDH值显著升高(P<0.05),SIRT1蛋白表达明显下降(P<0.05),提示SIRT1表达的缺失会加重MI损伤。结论:标本配穴电针可能是通过上调心肌细胞中的SIRT1的表达从而有效抗MI损伤。

Objective： To research the mechanism of manifestation points and root cause points combination in treating myocardial ischemia from the point of SIRT1 in cardiac muscle cells. Methods： fifty SPF Wistar rats were randomly divided into normal group, model group, treatment group（manifestation points and root cause points combination）, resveratrol group, and nicotinamide group, with 10 rats in each group. The rat model of myocardial ischemia was established by injection of isoprenaline hydrochloride; Before modeling, the rats in treatment group was treated with electro-acupuncture therapeutic apparatus of HANS-200 type at double Neiguan（PC6）, double Zusanli（ST36）, and Guanyuan（RN4） for 10 min, dilatational wave, 2^-100 Hz and 1m A; resveratrol group and nicotinamide group were respectively intragastric administrated with resveratrol solution and nicotinamide solution. After treatment, the change of S-T segment of ECG and myocardial enzyme activity were observed, the protein level of SIRT1 in cardiac muscle cells was detected by Western blot. Results： Compared with normal group, S-T section of ECG and the activity of myocardial enzyme activity in model group were significantly increased（P〈0.01）, the elevation range of S-T section is higher than 0.1m V, which means the myocardial ischemia model is successfully established（P〈0.01）; Compared with model group, S-T section of ECG and the activity of myocardial enzyme in both treatment group and resveratrol group were all decreased（P〈0.05）, which means both manifestation points and root cause points combination and resveratrol could prevent myocardial ischemia injury effectively. Compared with model group, the protein level of SIRT1 in nicotinamide group was lower（P〈0.05）, but the protein level of SIRT1 in resveratrol group was higher（P〈0.05）, which means nicotinamideand resveratrol could respectively inhibit and raise the protein level of SIRT1; There was no significant difference between the levels of protein expression of SIRT1 in treatment group and resveratrol group, which means manifestation points and root cause points combination could obviously rise the protein level of SIRT1. Conclusion： The manifestation points and root cause points combination could prevent myocardial ischemia injury effectively by improving SIRT1 level in cardiac muscle cells.