柔肝通络汤对MCAO大鼠自噬蛋白Beclin1及LC3表达的影响  

Effect of Rougan Tongluo Decoction on Beclin1 and LC3 expression of autophagy protein in MCAO rats

作　　者：龙华君[1,3] 杨颖 李映辰 胡国恒[4] 柏正平[3] 伍大华[3] LONG Hua-jun;YANG Ying;LI Ying-cheng;HU Guo-heng;Bai Zheng-ping;WU Da-hua(Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Loudi Hospital of Traditional Chinese Medicine,Loudi Hunan 417000,China;Affiliated Hospital of Hunan Research Institute of Tradition-al Chinese,Changsha 410006,China;First Hospital of Hunan University of Chinese Medicine,Changsha,410007,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南省娄底市中医医院,湖南娄底417000 [3]湖南省中医药研究院附属医院,湖南长沙410006 [4]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《时珍国医国药》2023年第8期1845-1849,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81904173);湖南中医药大学中西医结合一流学科开放基金(2020ZXYJH02);湖南省中医药科研计划项目重点课题(2021019);湖南省卫生健康委科研计划课题(C202303078160);赣南医学院心脑血管疾病防治教育部重点实验室开放基金(XN202011)。

摘　　要：目的从细胞自噬的角度探讨柔肝通络汤治疗脑缺血再灌注损伤的作用机制。方法将120只SD大鼠随机分为空白组、假手术组、模型组及柔肝通络汤组。空白组不予处理,假手术组及模型组予蒸馏水灌胃,柔肝通络汤组柔肝通络汤灌胃,均预处理1周。末次灌胃30min后,建立脑缺血再灌注模型。按再灌注3、6、12、24h时间点进行大鼠神经功能缺损评分并取脑,进行神经功能评分、TTC染色、HE染色、免疫组化法检测Beclin1、LC3蛋白的表达。结果与空白组及假手术组相比,模型组大鼠神经功能缺损评分升高(P<0.01),模型组与柔肝通络汤组均可见面积不等的梗死灶(P<0.01);模型组大鼠梗死侧脑组织细胞形态变性、坏死,出现噬神经细胞现象,且随时间增长而加重,Beclin1及LC3表达升高(P<0.01)。与模型组比较,柔肝通络汤组神经功能缺损评分下降(P<0.05),梗死面积有所减小,差异有统计学意义(P<0.05)。且柔肝通络汤组各亚组较模型组细胞形态损伤明显减轻,Beclin1及LC3表达均降低(P<0.01)。结论柔肝通络汤能改善大鼠神经功能缺损,缩小脑梗死灶,减轻神经细胞损伤。下调缺血再灌注损伤后自噬蛋白Beclin1、LC3的表达,减轻脑缺血再灌注损伤。Objective To explore the effect of Rougan Tongluo Decoction on cerebral ischemia reperfusion injury from the per⁃spective of autophagy mechanism.Methods 120 SD rats were randomly divided into 4 groups:blank group,sham operation group,model group and Rougan Tongluo Decoction group.The blank group was not treated,the sham operation group and the mod⁃el group were given distilled water gavage,and the Rougan Tongluo Decoction group was gavaged with high-dose Rougan Tongluo decoction,all of which were pretreated for one week.30 minutes after the last gavage,a cerebral ischemia-reperfusion model was established.Neurological impairment scores were performed at 3,6,12,and 24h time points of reperfusion and brains were taken to detect Beclin1 and LC3 protein expressions by nerve function scores,TTC staining,HE staining,and immunohistochemistry.Results Compared with blank group and sham group,the neurological deficit score of rats in model group was increased(P<0.01),and infarcts with different sizes were observed in both model group and Rougan Tongluo Decoction group(P<0.01).In the model group,the cerebral tissue of the infarct side of rats was morphologically degenerated and necrotic,and the phenomenon of nerve macrophage was observed,and the expression of Beclin1 and LC3 was increased with the increase of time(P<0.01).Compared with the model group,the neurological deficit score and infarct size of the Rougan Tongluo Decoction group were de⁃creased(P<0.05),and the difference was statistically significant(P<0.05).In addition,compared with model group,the morphological damage of cells in Rougan Tongluo Decoction group was significantly reduced,and the expressions of Beclin1 and LC3 were decreased(P<0.01).Conclusion Ruogan Tongluo Decoction can improve nerve function defect,narrow cerebral in⁃farction and reduce nerve cell damage in rats.The expressions of Beclin1 and LC3 were down-regulated after ischemia reperfusion injury to reduce cerebral ischemia reperfusion injury.

关 键 词：脑缺血再灌注损伤 柔肝通络汤 自噬 BECLIN1 LC3 

分 类 号：R285.5[医药卫生—中药学]