“双固一通”针法对急性脑缺血大鼠脑组织自噬相关基因表达的影响

The effect of“Shuanggu Yitong”acupuncture therapy on the expression of autophagy-related genes in brain tissue of rats with acute cerebral ischemia

目的观察"双固一通"针法对急性脑缺血大鼠自噬相关基因的影响,探讨其对急性脑缺血损伤的保护机制。方法将50只SD大鼠随机分为正常组、模型组、假手术组、穴位组、非穴位组,每组10只。采用开颅法电凝大脑中动脉制备急性脑缺血模型。穴位组针刺"足三里"、"肾俞"、"百会"、"水沟"穴,非穴位组取上述穴位旁开5mm非穴位处针刺。神经功能缺损评分(m NSS)测定大鼠神经功能损伤情况,2,3,5-三苯基氯化四氮唑(TTC)染色法检测脑梗死面积,干湿重法检测脑组织含水量,实时荧光定量PCR法检测Ⅱ型微管相关蛋白轻链3(LC3II)mRNA相对表达水平并计算LC3II/LC3I比值。结果模型组mNSS评分、脑梗死面积以及脑组织含水量明显高于正常组和假手术组(P<0.05);穴位组明显低于非穴位组和模型组(P<0.05)。模型组脑组织LC3II mRNA相对表达水平和LC3II/LC3I比值明显高于正常组和假手术(P<0.05);穴位组明显高于非穴位组和模型组(P<0.05);而假手术组与正常组之间各指标差异无统计学意义(P>0.05);非穴位组与模型组之间各指标差异无统计学意义(P>0.05)。结论"双固一通"针法能够显著改善急性脑缺血大鼠神经功能缺损,减少脑梗死面积,降低脑组织含水量,对急性脑缺血大鼠具有神经保护作用,其机制可能与"双固一通"针法激活细胞自噬有关。

Objective To observe the effect of"Shuanggu Yitong"acupuncture therapy on autophagy-related genes in rats with acute cerebral ischemia,and investigate the protective mechanism of"Shuanggu Yitong"acupuncture therapy on acute cerebral ischemic injury. Methods A total of 50 SD rats were randomized into a normal group,a sham model group,a model group,a sham acupuncture group,and an acupuncture group,with 10 rats in each group. The model of acute cerebral ischemia was established with craniotomy electrocoagulation to block the middle cerebral artery. Acupuncture was administered at the acupoints Zusanli(ST-36)and Shenshu(BL-23)Baihui(GV-20),Shuigou(GV-26) in acupuncture group. In SA group,acupuncture was performed at sham acupoints 5 mm away from ST-36 and BL-23,GV-20,GV-26 and other operations were the same as acupuncture group. The neural function deficit was detected by modified neurological severity score(m NSS). The brain infarct volume was detected by 2,3,5-triphenyltetrazoliumchloride(TTC)staining. The degree of brain edema was analyzed through dry/wet weight measurement. The relative expression ofmicrotubule-associated protein light chain 3-Ⅱ(LC3Ⅱ)mRNA was assessed by Real-Time Quantitative PCR,and calculate the ratio of LC3 II/LC3 I. Results In the aspect of neural function deficit,brain infarct volume and degree of brain edema,model group was significantly higher than normal group and sham model group(P<0.05);acupuncture group was significantly lower than sham acupuncture group and model group(P<0.05);in the aspect of the relative expression of LC3 II m RNA and the ratio of LC3 II/LC3,model group was significantly higher than normal group and sham model group(P<0.05);acupuncture group was significantly higher than sham acupuncture group and model group(P<0.05);the differences were not significant statistically in the comparison between normal group and sham model group(P>0.05);there was no statistically significant difference between sham acupuncture group and acupuncture group(P>0.05). Conclusion "Shuanggu Yitong"acupuncture therapy can significantly improve the neural function deficit,reduce the size of cerebral infarction and the degree of brain edema. Acupuncture has neuroprotective effect on acute cerebral ischemia in rats,which may be related to the activation of cell autophagy by"Shuanggu Yitong"acupuncture therapy.