针刺“百会”透“曲鬓”通过miR-34a-5p/KLF4信号通路治疗脑出血大鼠的机制研究  

Mechanism of Needling DU20 through GB7 in Treatment of Cerebral Hemorrhage in Rats Based on miR-34a-5p/KLF4 Signaling Pathway

作　　者：李丹 朱曦[3] 邓晓丰 李龙 LI Dan;ZHU Xi;DENG Xiaofeng;LI Long(Heilongjiang Mental Hospital,Harbin 150036,China;Fudan University,Shanghai 200433,China;The Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]黑龙江神志医院,黑龙江哈尔滨150036 [2]复旦大学,上海200433 [3]北京中医药大学第三附属医院,北京100029

出　　处：《针灸临床杂志》2022年第12期65-71,共7页Journal of Clinical Acupuncture and Moxibustion

基　　金：国家自然科学基金,编号:82174514;中国博士后科学基金,编号:2020M670940;黑龙江省博士后面上资助项目,编号:LBH-Z19034。

摘　　要：目的:探讨针刺“百会”透“曲鬓”通过miR-34a-5p/KLF4信号通路治疗脑出血大鼠的机制。方法:采用自体血注入尾壳核制取脑出血大鼠模型,将造模成功的60只大鼠随机平均分为假手术组(Sham组)、模型组(ICH组)、模型+针刺治疗组(ICH+acupuncture组)、模型+针刺治疗+agomir阴性对照组(ICH+acupuncture+agomir-NC组)与模型+针刺治疗+miR-34a-5p agomir组(ICH+acupuncture+miR-34a-5p agomir组),每组随机分1 d和7 d两个时间点。使用Longa评分评定大鼠神经功能缺损情况,应用Real-time PCR检测各组miR-34a-5p和KLF4表达,应用蛋白质印迹分析各组KLF4表达水平。结果:1 d和7 d,与ICH组比较,ICH+acupuncture组脑出血大鼠血肿周围miR-34a-5p表达降低,差异具有统计学意义(P<0.01),KLF4 mRNA表达、KLF4蛋白水平升高,差异具有统计学意义(P<0.01);与ICH+acupuncture组比,ICH+acupuncture+miR-34a-5p agomir组miR-34a-5p表达升高,差异具有统计学意义(P<0.01),KLF4 mRNA表达、KLF4蛋白水平降低,差异具有统计学意义(P<0.01);7 d时,与ICH组比较,ICH+acupuncture组脑出血大鼠Longa评分降低,差异具有统计学意义(P<0.01);与ICH+acupuncture组比,ICH+acupuncture+miR-34a-5p agomir组Longa评分升高,差异具有统计学意义(P<0.05)。结论:针刺“百会”透“曲鬓”可降低脑出血大鼠Longa评分分数,改善大鼠神经功能缺损的机制为通过miR-34a-5p/KLF4信号通路抑制miR-34a-5p表达、促进KLF4 mRNA表达和提高KLF4蛋白表达水平,减轻脑出血损害实现。Objective:To explore the mechanism of needling Baihui(DU20)through Qubin(GB7)in treatment of cerebral hemorrhage(ICH)in rats based on miR-34a-5p/KLF4 signaling pathway.Methods:ICH model was established by autologous blood injection into caudate putamen.60 successful modeling rats were randomly divided into the sham operation group,the ICH group,the ICH plus acupuncture group,the ICH and acupuncture plus agomir-NC group,as well as the ICH and acupuncture plus miR-34a-5p agomir group.All groups were sub divided into groups of 1 d and 7 d.Neurological defect was assessed by Longa method.mRNA expressions of miR-34a-5p and KLF4 were detected by Real-time PCR,and protein expression of KLF4 was analyzed by Western Blot in each group.Results:Compared with those in the ICH group,the expression of miR-34a-5p was decreased(P<0.01),and mRNA and protein expressions of KLF4 were increased in the ICH plus acupuncture group at the time points of 1 d and 7 d(P<0.01).Compared with those in the ICH plus acupuncture group,the expression of miR-34a-5p was increased(P<0.01),the mRNA and protein expressions of KLF4 were decreased in the ICH and acupuncture plus miR-34a-5p agomir group(P<0.01).Longa score of the ICH plus acupuncture group was lower than that of the ICH group at 7 d(P<0.01).Longa score of the ICH and acupuncture plus miR-34a-5p agomir group was higher than that of the ICH plus acupuncture group(P<0.05).Conclusion:Needling DU20 through GB7 can reduce Longa score and improve neurological defect in rats with cerebral hemorrhage by inhibiting the expression of miR-34a-5p,promoting the mRNA and protein expressions of KLF4,thus to reduce cerebral hemorrhage damage.

关 键 词：脑出血 针刺 miR-34a-5p KLF4 

分 类 号：R246.6[医药卫生—针灸推拿学]