化痰通络汤对脑缺血/再灌注大鼠“肠-脑”轴的干预作用  

Effect of Huatan Tongluo Decoction on Gut-Brain Axis in Rats with Cerebral Ischemia/Reperfusion

作　　者：唐三 杨筱倩[1] 朱秋燕 丁煌[2] 刘晓丹[2] 曾鑫冰 陆展辉 杨芙蓉 王宪庆[4] 黄小平[1,2] TANG San;YANG Xiao-qian;ZHU Qiu-yan;DING Huang;LIU Xiao-dan;ZENG Xin-bing;LU Zhan-hui;YANG Fu-rong;WANG Xian-qing;HUANG Xiao-ping(Laboratory of Vascular Biology,Hunan University of Chinese Medicine,Changsha 410208,China;Key Laboratory of Cardio-Brain Disease Prevention and Treatment of Integrated Traditional Chinese and Western Medicine,Hunan Provincial Key Laboratory of Cell Biology and Molecular Technology,Hunan University of Chinese Medicine,Changsha 410208,China;College of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Changde Vocational and Technical College,Changde 415000,China)

机构地区：[1]湖南中医药大学血管生物学实验室,长沙410208 [2]湖南中医药大学中西医结合心脑疾病防治湖南省重点实验室,细胞生物学与分子技术湖南省高校重点实验室,长沙410208 [3]湖南中医药大学中西医结合学院,长沙410208 [4]常德职业技术学院,常德415000

出　　处：《中国实验方剂学杂志》2021年第18期32-41,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81573875);湖南省高校创新平台开放基金项目(18K067);湖南中医药大学“十三五”一级学科基础医学建设项目(06);湖南省研究生优秀教学团队(基础医学研究生教学团队)项目([2019]370-118)。

摘　　要：目的:基于"肠-脑"轴探讨化痰通络汤(HTTLD)对大鼠脑缺血再灌注后脑组织和肠道形态和功能的影响。方法:将60只SPF级雄性大鼠随机分为假手术组,模型组,HTTLD高、中、低剂量组(28.66,14.33,7.16 g·kg^(-1)),依达拉奉(4 g·kg^(-1))+丁酸梭菌(5.0×10^(8)cfu·mL^(-1))组,采用大鼠局灶性脑缺血再灌注模型,灌胃给药HTTLD,神经功能缺损评分和2,3,5-氯化三苯四氮唑(TTC)染色法测定脑组织损伤,小肠推进率观察脑缺血再灌注对肠道动力的影响,十二指肠黏膜的病理形态学检测评价肠黏膜细胞损伤,酶联免疫吸附测定法(ELISA)测定血清D-乳酸(D-LAC),二胺氧化酶(DAO),细菌内毒素(LPS)含量,蛋白免疫印迹法(Western blot)检测十二指肠的闭合蛋白(Occludin),紧密连接蛋白-5(Claudin-5),闭锁小带蛋白-1(ZO-1)蛋白的表达。结果:脑缺血再灌注后,大鼠出现神经功能缺损症状,与假手术组比较,模型组神经功能缺损评分显著升高(P<0.01);与模型组比较,HTTLD中、高剂量组能显著减少神经功能缺损症状,降低神经功能缺损评分(P<0.01)。与假手术组比较,模型组大鼠脑组织出现明显梗死灶,脑梗死率显著升高(P<0.01);与模型组比较,各给药组脑梗死体积明显减少(P<0.01),以HTTLD高剂量组的效应最佳,且效应具有剂量依赖性。小肠推进率结果显示,与假手术组比较,模型组小肠推进率显著降低(P<0.01);与模型组比较,各给药组能显著升高小肠推进率(P<0.01),以HTTLD高剂量效应最佳,且效应具有剂量依赖性。十二指肠黏膜组织病理学检测显示,脑缺血再灌注后十二指肠黏膜可见明显损伤,各给药组黏膜损伤情况明显减轻。与假手术组比较,模型组ZO-1,Occludin,Claudin-5蛋白表达均显著降低(P<0.01);与模型组比较,HTTLD低、中、高剂量组能不同程度上调ZO-1,Occludin,Claudin-5蛋白表达(P<0.05,P<0.01),以高剂量组的效应最为显著。与假手术组比较,模型组血清DLAC,DAObjective: To explore the effects of Huatan Tongluo Decoction(HTTLD) on the morphology and function of brain tissues and intestine in rats with cerebral ischemia/reperfusion based on the gutbrain axis. Method: Sixty SPF male rats were randomly divided into a sham operation group,a model group,high-(28.66 g·kg^(-1)),medium-(14.33 g·kg^(-1)),and low-dose(7.16 g·kg^(-1))HTTLD groups,and an edaravone(4 g·kg^(-1))+Clostridium butyricum(5.0×10^(8) cfu·mL^(-1))group. The model was established by focal cerebral ischemia/reperfusion in rats. The drugs were administered by gavage. The brain tissue injury was determined by neurological deficit score and 2,3,5-triphenyl tetrazolium chloride(TTC)staining. The effect of cerebral ischemia/reperfusion on intestinal motility was assessed by the propulsion rate of small intestine. The intestinal mucosal cell damage was evaluated by the pathomorphological examination of the duodenal mucosa. Enzymelinked immunosorbent assay(ELISA)was used to determine the content of D-lactate(D-LAC),diamine oxidase(DAO),and bacterial endotoxin(lipopolysaccharide,LPS)in serum. Western blot was used to detect the expression of Occludin,Claudin-5,and zonula occludens 1(ZO-1)in the duodenum. Result: After cerebral ischemia/reperfusion,rats developed neurological deficit symptoms. The neurological deficit score in the model group was higher than that in the sham operation group(P<0.01). Compared with the model group,the high-and medium-dose HTTLD groups could relieve the symptoms of neurological deficits and lower neurological deficit scores(P<0.01). The results of TTC staining showed that the model group presented obvious infarcts in brain tissues compared with the sham operation group(P<0.01). The cerebral infarction volumes of HTTLD groups were reduced compared with that in the model group(P<0.01),especially the highdose HTTLD group,and the effect was dose-dependent. Furthermore,the propulsion rate of small intestine in the model group was significantly reduced compared with that in the sham ope

关 键 词：“肠-脑”轴 脑缺血再灌注 化痰通络汤 肠道动力 肠道屏障 

分 类 号：R2-0[医药卫生—中医学] R22