针刺对出血性中风大鼠脑组织自噬相关蛋白表达的影响

Penetrative needling improves neurological function by up-regulating expression of autophagy related protein LC3 in rats with hemorrhagic stroke

目的:观察针刺'百会'透'曲鬓'穴对出血性中风大鼠自噬相关蛋白的影响,探讨针刺促进脑出血后大鼠神经功能恢复的机制。方法:120只SD大鼠随机分成假手术组、模型组、非穴位组、针刺组、雷帕霉素组,每组按脑出血后不同时间点(造模后第3、7天)分2个亚组。将50μL自体血注入到大鼠尾状核制备出血性中风模型。各干预组分别给予针刺'百会'透患侧'曲鬓'、非穴位针刺、侧脑室注射雷帕霉素进行干预。于各时间点对大鼠进行神经功能行为学评分(悬线测试、平衡行走测试)。评分结束后取各组大鼠出血半暗带区脑组织,免疫组织化学法检测微管相关蛋白1轻链3(LC3)蛋白的阳性表达,Western blot法检测LC3-Ⅱ/Ⅰ的变化。结果:与假手术组比较,模型组造模后第3、7天的神经功能行为学评分(悬线测试、平衡行走测试)显著降低(P<0. 05);与模型组比较,针刺组、雷帕霉素组大鼠神经功能行为学评分明显升高(P<0. 05),且针刺组高于雷帕霉素组(P<0. 05)。与同时间点假手术组比较,造模后第3、7天模型组大鼠脑组织LC3蛋白阳性表达、LC3-Ⅱ/Ⅰ明显升高(P<0. 05);与同时间点的模型组比较,针刺组大鼠脑组织LC3蛋白阳性表达、LC3-Ⅱ/Ⅰ明显升高(P<0. 05);与针刺组比较,雷帕霉素组大鼠脑组织LC3蛋白阳性表达、LC3-Ⅱ/Ⅰ明显升高(P<0. 05)。结论:针刺能上调出血性中风大鼠脑组织LC3-Ⅱ/Ⅰ,良性激活自噬水平,进而改善大鼠神经功能缺损。

ObjectiveTo observe the effect of penetrative needling from'Baihui'(GV20)to'Qubin'(GB7)on neurological function and expression of autophagy related protein microtubule-associated protein,light chain 3(LC3)in rats with hemorrhagic stroke,so as to explore its mechanism underlying improvement of hemorrhagic stroke.MethodsA total of 120 male SD rats were randomly divided into sham operation,model,non-acupoint,acupuncture and medication(Rapamycin)groups which were further divided into two time-point subgroups(3,7 days after modeling,n=12/subgroup).The intracerebral hemorrhage model was established by injection of the rat’s auto-blood(50μL)into the putaman region.Penetrative needling from GV20 to GB7 or sham acupoints(about 1 cm beside GV20 and GB7)was conducted for 30 min,twice daily for 7 days.For rats of the medication group,Rapamycin solution(7μmol/L)was injected into the right lateral ventricle.The neurological functions(locomotor and balance deficits)were evaluated according to suspended wire test(0-6 points)and horizontal board walking test(0-6 points).Immunohistochemistry and Western blot were used to detect the expression of total LC3 and expression of LC3-Ⅰand LC3-Ⅱproteins in the ischemic penumbra region of brain tissue,respectively.ResultsAfter modeling,the neurological function scores were significantly decreased on day 3 and 7,the levels of LC3-Ⅱ/Ⅰand LC3 protein on day 3 and 7 were significantly increased in the model group relevant to the sham operation group(P<0.05).Following the interventions,the neurological function scores as well as LC3-Ⅱ/Ⅰand LC3 protein expression were significantly increased in both acupuncture and medication groups compared with the model group(P<0.05).The effect of Rapamycin was obviously stronger than that of penetrative needling in up-regulating the expression of LC3-Ⅱ/Ⅰand LC3 protein(P<0.05).ConclusionPenetrative needling can improve neurologic function in hemorrhagic stroke rats,which is related with its effect in up-regulating the expression of autophagy-related protein LC3.