扎冲十三味丸调控Hippo信号通路对脑缺血大鼠的影响  

Effects of Zhachong Shisanwei Pills on Rats with Cerebral Ischemia by Regulating Hippo Signaling Pathway

作　　者：理扎·沙布尔江 张晓璐 尚津锋 王璟邑 闫明雪 宋琪 文胤琏 黄贵金凤 陈文彬 白梅荣[2] 刘欣[1] Lizha Shabuerjiang;ZHANG Xiaolu;SHANG Jinfeng;WANG Jingyi;YAN Mingxue;SONG Qi;WEN Yinlian;HUANG Guijinfeng;CHEN Wenbin;BAI Meirong;LIU Xin(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Key Laboratory of Mongolian Medicine Research and Development Engineering,Ministry of Education,Inner Mongolia Minzu University,Tongliao 028000,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]内蒙古民族大学蒙医药研发工程教育部重点实验室,内蒙古通辽028000

出　　处：《中国中医药信息杂志》2024年第11期96-103,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：蒙医药研发工程教育部重点实验室开放课题(MDK2023062)。

摘　　要：目的观察扎冲十三味丸对脑缺血大鼠的影响及作用机制。方法将75只大鼠随机分为假手术组、模型组、阳性药组(金纳多,21.6 mg/kg)和扎冲十三味丸低、中、高剂量组(81、162、324 mg/kg),各给药组分别予相应药物灌胃5 d,第6日线栓法制备脑缺血大鼠模型,造模后24 h继续以相同方式给药2 d。对大鼠进行神经功能评分、横木行走评分、抓握力测试,TTC染色检测脑梗死率,HE染色、尼氏染色观察脑组织形态,TUNEL染色检测脑组织细胞凋亡率,通过转录组学筛选扎冲十三味丸治疗脑缺血差异基因,RT-qPCR、免疫组化和Western blot检测差异基因mRNA和蛋白表达。结果与假手术组比较,模型组大鼠神经功能评分、横木行走评分、抓握力降低,脑梗死率升高,神经元核固缩、空泡样改变,细胞间隙增宽,尼氏体数量减少,细胞凋亡率增加(P<0.01,P<0.001);与模型组比较,扎冲十三味丸中剂量组大鼠神经功能评分、横木行走评分、抓握力升高,脑梗死率降低,神经元损伤程度较低,尼氏体数量增加,细胞凋亡率降低(P<0.05,P<0.01)。转录组学和生物信息学分析筛选出扎冲十三味丸抗脑缺血的相关通路Hippo信号通路。该通路关键基因哺乳动物不育系20样激酶(MST1)1、Yes相关蛋白(YAP)1、大肿瘤抑制激酶(LATS)1、TEA域家族成员(TEAD)1检测结果显示,模型大鼠脑组织MST1 mRNA和蛋白表达显著升高,YAP1、LATS1、TEAD1 mRNA和蛋白表达显著降低;扎冲十三味丸可下调模型大鼠脑组织MST1表达,上调YAP1、LATS1、TEAD1表达。结论扎冲十三味丸可能通过Hippo信号通路发挥抗脑缺血作用。Objective To investigate the effects and mechanism of Zhachong Shisanwei Pills on rats with cerebral ischemia.Methods Totally 75 rats were randomly divided into sham-operation group,model group,positive drug group(Ginaton,21.6 mg/kg),and Zhachong Shisanwei Pills low-,medium-,and high-dosage groups(81,162,324 mg/kg).Each treatment group was given the corresponding drug by gavage for 5 days.On the 6th day,a cerebral ischemia rat model was prepared by suture method.After 24 hours of modeling,the drugs were given in the same manner for 2 days.Neurological function scoring,horizontal beam walking scoring,and grip strength testing were performed on rats.TTC staining was used to detect the cerebral infarction rate,HE staining and Nissl staining were used to observe the morphology of brain tissue.TUNEL staining was used to detect the apoptosis rate of brain tissue cells.Differential genes in the treatment of cerebral ischemia using Zhachong Shisanwei Pills were screened by transcriptomics,and RT-qPCR,immunohistochemistry and Western blot were used to detect differential gene mRNA and protein expression.Results Compared with the sham-operation group,the model group rats showed a decrease in neurological function scores,horizontal beam walking scores,grip strength,an increase in cerebral infarction rate,neuronal nucleus condensation,vacuolar changes,widened intercellular spaces,the number of Nissl bodies reduced,and the apoptosis rate increased(P<0.01,P<0.001);compared with the model group,the Zhachong Shisanwei Pills medium-dosage group showed an increase in neurological function score,horizontal beam walking score,and grip strength in rats,a decrease in cerebral infarction rate,a lower degree of neuronal damage,an increase in the number of Nissl bodies,and a decrease in cell apoptosis rate(P<0.05,P<0.01).Transcriptome and bioinformatics analysis screened the Hippo signaling pathway related to the anti-cerebral ischemia effect of Zhachong Shisanwei Pills.The key genes of this pathway,mammalian sterile line 20 like kinase(

关 键 词：脑缺血 扎冲十三味丸 转录组学 Hippo信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]