目的 基于miR-34家族基因与Kruppel样因子4（Kruppel-like factor 4，KLF4）/血管内皮生长因子（vascular endothelial growth factor，VEGF）传导轴的相关性探讨活血荣络方促脑梗死后血管新生的机制。方法 将SD大鼠随机分为对照组、模型组、活血荣络方（11.7 g/kg）组和丁苯酞（60 mg/kg）组，每组10只，建立大脑中动脉闭塞/再灌注（middle cerebral artery occlusion/reperfusion，MCAO/R）模型，连续ig药物7 d。采用改良神经功能缺损评分（modified neurological severity score，mNSS）评价大鼠神经功能缺损症状；采用免疫荧光染色观察海马区CD34及皮质区VEGF表达；采用Western blotting法检测各组大鼠梗死侧脑组织中KLF4和VEGF蛋白表达情况；采用qRT-PCR法检测各组大鼠脑组织中miR-34家族基因的表达；采用Pearson相关分析探讨miR-34家族基因与mNSS、KLF4、VEGF的相关性。结果 活血荣络方能够显著改善大鼠神经功能缺损症状（P<0.01），增加海马区CD34阳性细胞及皮质区VEGF表达（P<0.01），增强脑组织中KLF4和VEGF蛋白表达（P<0.01），抑制miR-34家族基因miR-34a、miR-34b、miR-34c mRNA表达（P<0.01）；且mNSS与miR-34a、miR-34b、miR-34c呈高度正相关（P<0.05）；VEGF与miR-34a、miR-34b、miR-34c呈高度负相关（P<0.05）；KLF4与miR-34b呈高度负相关（P<0.05），与miR-34c呈高度负相关（P<0.01）。结论 活血荣络方能够抑制miR-34a、miR-34b、miR-34c mRNA表达，激活KLF4/VEGF传导轴，促进脑梗死后血管新生，进而改善神经功能缺损症状。

Objective Based on the correlation between miR-34 family genes and Kruppel-like factor 4 (KLF4)/vascular endothelial growth factor (VEGF) conduction axis to investigate the mechanism of Huoxue Rongluo Recipe (活血荣络方) on promoting angiogenesis after cerebral infarction. Methods SD rats were randomly divided into control group, model group, Huoxue Rongluo Recipe (11.7 g/kg) group and butylphthalide (60 mg/kg) group, 10 rats in each group. Middle cerebral artery occlusion/reperfusion (MCAO/R) rats model were established, and rats were ig drugs for 7 d. Modified neurological severity score (mNSS) was used to evaluate the symptoms of neurological deficit in rats; Immunofluorescence staining was used to observe the expression of CD34 in hippocampus and VEGF in cortex; Western blotting was used to detect the expression of KLF4 and VEGF in infarcted brain tissue; qRT-PCR was used to detect the expression of miR-34 family genes in brain tissue; Pearson correlation analysis was used to observe the correlation between miR-34 family genes and mNSS, KLF4 and VEGF. Results Huoxue Rongluo Recipe significantly improved the symptoms of neurological deficits in rats (P < 0.01), increased expression of CD34 positive cells in hippocampus and expression of VEGF in cortex (P < 0.01), enhanced expression of KLF4 and VEGF protein in brain tissue (P < 0.01), inhibited mRNA expressions of miR-34a, miR-34b and miR-34c (P < 0.01); mNSS was highly positively correlated with miR-34a, miR-34b and miR-34c (P < 0.05); VEGF was highly negatively correlated with miR-34a, miR-34b and miR-34c (P < 0.05); KLF4 was highly negatively correlated with miR-34b (P < 0.05), highly negatively correlated with miR-34c (P < 0.01). Conclusion Huoxue Rongluo Recipe can inhibit expressions of miR-34a, miR-34b and miR-34c, activate KLF4/VEGF conduction axis, promote angiogenesis after cerebral infarction, and then improve the symptoms of neurological deficits.

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国家自然科学基金资助项目（81874463）；湖南省科技厅科技创新平台与人才计划项目（2017SK4005）；湖南省重点研发计划项目（2020SK2092）；湖南省财政中医药项目名院工程（rsk-010-013/006-09）；湖南省教育厅一般项目（20C1405）；湖南省卫健委科研项目（202103071190）；湖南省中医药管理局资助项目（202038，202046，2021218）；湖南省研究生创新项目（CX20190587）；湖南中医药大学中西医结合一流学科开放基金项目（2019ZXYJH08，2018ZXYJH10）；湖南中医药大学研究生培养质量工程专项（2020CX62）