基于NLRP3/Caspase-1信号通路研究三叶香茶菜抗小鼠CCL4急性肝损伤机制  

Isodon Ternifolius(D.Don)Kudo Improves Acute Liver Injury Induced by CCL_(4) in Mice Through Inhibiting NLRP3/Caspase-1 Signaling Pathway

作　　者：周至品 农汝楠 覃乐 王竟静 李秋甫 吴妍 刘代华 周蓓 Zhou Zhipin;Nong Runan;Qin Le;Wang Jingjing;Li Qiufu;Wu Yan;Liu Daihua;Zhou Bei(Liuzhou People9 s Hospital of Guangxi Medical University,Liuzhou 545006,China;School of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530001,China)

机构地区：[1]广西医科大学附属柳州市人民医院,柳州545006 [2]广西中医药大学药学院,南宁530001

出　　处：《世界科学技术-中医药现代化》2021年第12期4678-4685,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会地区科学基金项目(81760751):基于TLR4/NF-κB信号通路研究三叶香茶菜抗肝纤维化作用机制,负责人:周至品;广西壮族自治区中医药管理局自筹经费课题(GZZC2019110):基于NLRP3炎症小体研究三叶香茶菜抗小鼠CCL4急性肝损伤作用机制,负责人:周至品;柳州市科学技术局科技计划项目(2020NBAB0815):三叶香茶菜介导miRNA-214-5p靶向调控TGF-β1/Smads信号通路抗肝纤维化机制研究,负责人:周至品。

摘　　要：目的研究三叶香茶菜对四氯化碳(carbon tetrachloride,CCL4)急性肝损伤小鼠NLRP3/casepase-1信号通路的影响。方法采用CCL_(4)小鼠急性肝损伤动物模型,随机分为模型组,正常组,联苯双脂组,三叶香茶菜组分别为高(150 g·kg^(-1))、中(75 g·kg^(-1))、低(37.5 g·kg^(-1))剂量组(按生药量计)。各组干预7天后,于第7天晚上注射CCL4油溶液诱导急性肝损小鼠动物模型(除正常组外)。12 h后,称量各组肝、脾重量,并计算肝、脾指数,生化法检测各组小鼠血清中谷丙转氨酶(ALT)活力、谷草转氨酶(AST)活力,ELISA法检测小鼠血清半胱肽氨酸蛋白酶-1(Caspase-1)、白细胞介素-1β(interleukin-1β,IL-1β)水平及肝组织NLR家族含有Pyrin结构域的蛋白质3(NLR Family,Pyrin Domain Containing Protein 3,NLRP3)含量。同时,采用免疫组化检测肝组织中NLRP3含量。取肝组织制成病理切片,苏木精-伊红染色法(hematoxylin-eosin staining,HE)染色后观察肝组织细胞形态学变化,并评分。结果(1)各组肝指数与模型组比较无明显变化(P<0.05);与模型组比较,正常组、联苯双脂组、三叶香茶菜低剂量组脾指数均显著下降(P<0.05)。(2)与正常组对比,模型组小鼠血清ALT、AST、Caspase-1、IL-1β以及小鼠肝组织NLRP3显著升高(P<0.05或P<0.01);与模型组比较,各组小鼠血清ALT、AST、Caspase-1、IL-1β以及小鼠肝组织NLRP3显著下降(P<0.05或P<0.01);但模型组与三叶香茶菜低剂量组对比,小鼠血清ALT、Caspase-1、IL-1β无显著性差异(P<0.05)。(3)光镜下观察发现,与模型组相比,三叶香茶菜高、中、低剂量组小鼠肝组织损伤明显减少,肝组织细胞坏死细胞减轻,有少量炎性浸润,各组病理评分差异具显著性差异(P<0.05)。结论三叶香茶菜对CCL_(4)诱导急性肝损伤小鼠具有保护作用,其护肝作用机制可能是是通过抑制NLRP3/Caspase-1信号通路的活化,减少炎症介质释放,从而抑制肝组织炎症损害。Objective To investigate that Isodon ternifolius(D.Don)Kudo protects mice from acute liver injury induced by carbon tetrachloride(CCL_(4))by affecting the NLRP3/Caspase;signaling pathway.Methods In this study,we used CCL_(4)to induce acute liver injury in mice and randomly divided them into model group,normal group,bifendate group,and Isodon ternifolius(D.Don)Kudo group,which were set as high(150 g·kg^(-1)),medium(75 g·kg^(-1)),and low(37.5 g·kg^(-1))dose groups(the dose of raw drug).After 7 days of our experimental intervention,each group(except the normal group)was injected with CCL_(4)oil solution to induce acute liver injury in mice.After 12 hours,we measured the weight of liver and spleen in each group,and calculated the index of liver and spleen.The activity of alanine aminotransferase(ALT),aspartate aminotransferase(AST)was tested by biochemical method.Caspase-1,interleukin-1beta(IL-1β)and the content of NLR Family,Pyrin Domain Containing Protein 3(NLRP3)was tested by ELISA method.Meanwhile,NLRP3 was detected by immunohistochemistry.The liver tissues were taken and made into pathological sections.After hematoxylin-eosin staining(HE),the morphological changes of liver cells were observed and scored.Results(1)There was no significant change in liver index between each group and model group(P<0.05).Compared with the model group,the spleen index of the normal group,the bifendate group,and the low dose group of Isodon ternifolius(D.Don)Kudo decreased significantly(P<0.05).(2)Compared with the normal group,serum ALT,AST,Caspase-1,IL-1βand liver NLRP3 in the model group were significantly increased(P<0.05 or P<0.01).Compared with the model group,serum ALT,AST,Caspase-1,IL-1beta and liver NLRP3 of mice in each intervention group were significantly decreased(P<0.05 or P<0.01).However,there was no significant difference in serum ALT,Caspase-1 and IL-1beta between the model group and the low-dose group(P<0.05).(3)Light microscopic observation showed that compared with the model group,the mice in the high,medium

关 键 词：三叶香茶菜 NLRP3/Caspase-1信号通路 急性肝损伤 

分 类 号：R285.5[医药卫生—中药学]