益气活血化浊解毒方调节脂多糖和TLR4/MyD88/MAPK信号通路减轻大鼠脑缺血再灌注损伤研究  

Study of Yiqi Huoxue Huazhuo Jiedu Formula on reducing cerebral ischemia reperfusion injury in rats by regulating lipolysaccharides and TLR4/MyD88/MAPK signalingpathway

作　　者：孙亚萍 霍瑞卿 赵敏菡 石瑞 孙玲玲 谢占维 田军彪[2] SUN Yaping;HUO Ruiqing;ZHAO Minhan;SHI Rui;SUN Lingling;XIE Zhanwei;TIAN Junbiao(Hebei University of Chinese Medicine,Shijiazhuang 050091,China;Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China)

机构地区：[1]河北中医药大学,石家庄050091 [2]河北省中医院,石家庄050011

出　　处：《中华中医药杂志》2023年第12期5997-6002,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河北省重点研发计划项目(No.20377710D),河北省中医药管理局科研计划重点项目(No.Z2022008);河北中医学院科研能力提升重点项目(No.KTZ2019028);河北中医学院研究生创新资助项目(No.XCXZZBS2022008)。

摘　　要：目的:观察益气活血化浊解毒方对脑缺血再灌注损伤大鼠脂多糖(LPS)及TLR4/MyD88/MAPK信号通路的影响,并初步探讨其作用机制。方法:将60只SPF级雄性SD大鼠随机分为假手术组(SHAM组),模型组(MCAO组),模型+LPS组(MCAO+LPS组),益气活血化浊解毒方低、中、高剂量组(简称低、中、高剂量组)。采用线栓法制备大鼠大脑中动脉闭塞再灌注模型,3d后观察大鼠神经功能缺损评分和脑梗死面积,HE染色观察脑和结肠组织病理改变,ELISA检测脑和结肠组织LPS含量,RT-PCR检测脑组织TLR4、MyD88mRNA表达,WesternBlot检测脑组织TLR4、MyD88、P-p38MAPK、p38MAPK、p-ERK、ERK、p-JNK、JNK、COX-2、iNOS蛋白表达。结果:与SHAM组比较,MCAO组与MCAO+LPS组神经功能缺损严重,脑梗死面积明显增加,脑和结肠组织LPS含量显著增加,脑组织TLR4、MyD88、P-p38MAPK/p38MAPK、p-ERK/ERK、p-JNK/JNK、COX-2、iNOS表达显著增加(P<0.05)。与MCAO组比较,各剂量组神经功能缺损症状减轻,脑梗死面积显著减少(P<0.05);中、高剂量组脑和结肠组织LPS含量显著降低(P<0.05),脑组织TLR4、MyD88、COX-2,iNOS、P-p38MAPK/p38MAPK、p-JNK/JNK表达显著降低(P<0.05);高剂量组脑组织p-ERK/ERK表达显著降低(P<0.05)。结论:益气活血化浊解毒方对脑缺血再灌注损伤大鼠具有保护作用,其机制可能与调节LPS含量,抑制TLR4/MyD88/MAPK信号通路的激活,减轻炎症反应密切相关.Objective:To observe the effect of Yiqi Huoxue Huazhuo Jiedu Formula(YHHJF)on expression of lipolysaccharides(LPS)and TLR4/MyD88/MAPK signaling pathway in rats with cerebral ischemia reperfusion injury(CIRI),and preliminarily explore the mechanism.Methods:A total of 60 male SD rats of SPF grade were randomly divided into sham-operation group(SHAM group),model group(MCAO group),model+lipolysaccharides group(MCAO+LPS group),YHHJF low/medium/high dose group(TCM-L,TCM-M,TCM-H group).The middle cerebral artery occlusion model was prepared by suture method.The neurological function defects and infarct size of rats were detected 3 days after administration.The pathological changes in brain and colon tissue were observed by HE,the content of LPS in brain and colon tissue were detected by ELISA.The mRNA expression of TLR4,MyD88 in brain tissue were studied by RT-PCR,the protein expression of TLR4,MyD88,p-p38MAPK,p38MAPK,p-ERK,ERK,p-JNK,JNK,COX-2 and iNOS in brain tissue were studied by Western Blot.Results:Compared with SHAM group,the neural function impairment was aggravated in model and model+LPS groups,the infarct size was markedly increased,the content of LPS in brain and colon tissue were increased,the expression of TLR4,MyD88,P-p38MAPK/p38MAPK,p-ERK/ERK,p-JNK/JNK,COX-2,iNOS were increased significantly(P<0.05).Compared with MCAO group,the neurological function impairment and infarct size were markedly decreased in TCM-L/TCM-M/TCM-H group(P<0.05),the content of LPS in brain and colon tissue were decreased in TCM-M/TCM-H group(P<0.05),the expression of TLR4,MyD88,P-p38MAPK/p38MAPK,p-JNK/JNK,COX-2,iNOS were decreased significantly in TCM-M/TCM-H group(P<0.05),the expression of p-ERK/ERK was decreased significantly in TCM-H group(P<0.05).Conclusion:YHHJF exerts a strong protective effect on CIRI rats by regulating the content of LPS,inhibiting the activation of TLR4/MyD88/MAPK signaling pathway and alleviating inflammatory response.

关 键 词：益气活血化浊解毒方 脑缺血再灌注损伤 TLR4/MyD88/MAPK信号通路 脂多糖 炎症 

分 类 号：R285.5[医药卫生—中药学]