解毒化瘀颗粒调控ROS-NLRP3信号通路缓解肝衰竭大鼠肝脏炎症应激的实验研究  

Experimental study on Jiedu Huayu Granules regulating ROS-NLRP3 signaling pathway to alleviate liver inflammatory stress in rats with liver failure

作　　者：吴智鹏 张荣臻[2] 王明刚[2] 张钰琴 王秀峰[2] WU Zhi-peng;ZHANG Rong-zhen;WANG Ming-gang;ZHANG Yu-qin;WANG Xiu-feng(Graduate School of Guangxi University of Chinese Medicine,Nanning,530200,China;First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Department of Cell Biology and Genetics,School of Pre-clinical Medicine,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西中医药大学研究生院,广西南宁530200 [2]广西中医药大学第一附属医院,广西南宁530023 [3]广西医科大学基础医学院细胞生物学与遗传学教研室,广西南宁530021

出　　处：《时珍国医国药》2024年第5期1025-1029,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(82160881);广西自然科学基金(2018GXNSFGA281002);广西中医药大学博士启动基金(2020BS026);广西中医药大学桂派中医药传承创新团队项目(2022A001);广西中医药大学第一附属医院博士启动基金(2020BS003)。

摘　　要：目的分析解毒化瘀颗粒(JDHY)通过调控ROS-NLRP3信号通路改善肝衰竭大鼠肝脏炎症反应的相关分子机制。方法将36只雄性Wistar大鼠随机分为空白组(Control)、模型组(Model)、解毒化瘀颗粒组(JDHY),每组各12只。采用D-氨基半乳糖(D-GalN)联合脂多糖(LPS)一次性腹腔注射建立急性肝衰竭大鼠模型。JDHY组在造模前给予JDHY(57.55 g·kg^(-1)·d^(-1)),连续灌胃72h并延续至造模后24h。造模后24h处死各组大鼠,收集血清及组织样本,采用全自动生化分析仪测定血清ALT、AST和TBIL水平,ELISA测定血清IL-1β和IL-18,评估ROS生成,免疫组织化学分析肝组织NLRP3和GSDMD表达,蛋白质印迹分析肝组织NLRP3、GSDMD、GSDMD-N、ASC和Caspase-1表达。结果JDHY能有效改善肝衰竭大鼠肝功能(ALT、AST、TBIL)(P<0.05),减少ROS过度产生(P<0.05)和炎症因子(IL-1β、IL-18)释放。此外,与模型组比较,JDHY降低了肝衰竭大鼠肝组织中NLRP3、GSDMD、GSDMD-N、ASC和Caspase-1的表达(P<0.05)。结论JDHY能抑制ROS-NLRP3信号通路过度激活、减少促炎因子释放,缓解肝脏炎症应激水平从而拮抗肝衰竭。Objective To analyze the molecular mechanism of Jiedu Huayu Granules(JDHY)in improving liver inflammation in rats with liver failure by regulating ROS-NLRP3 signaling pathway.Methods Thirty-six male Wistar rats were randomly divided into control group,model group and JDHY group,with 12 rats in each group.A rat model of acute liver failure was established by intraperitoneal injection of D-galactosamine(D-GalN)combined with lipopolysaccharide(LPS).JDHY group was given JDHY(57.55 g·kg^(-1)·d^(-1))before modeling,continuous intragastric administration for 72 h and continued to 24 h after modeling.Rats in each group were sacrificed 24 h after modeling.Serum and tissue samples were collected.Serum ALT,AST and TBIL levels were measured by automatic biochemical analyzer.Serum IL-1βand IL-18 were measured by ELISA.ROS production was evaluated.The expression of NLRP3 and GSDMD in liver tissue was analyzed by immunohistochemistry.The expression of NLRP3,GSDMD,GSDMD-N,ASC and Caspase-1 in liver tissue was analyzed by Western blotting.Results JDHY could effectively improve liver function(ALT,AST,TBIL)(P<0.05),reduce the excessive production of ROS(P<0.05)and the release of inflammatory factors(IL-1β,IL-18)in rats with liver failure.In addition,compared with the model group,JDHY reduced the expression of NLRP3,GSDMD,GSDMD-N,ASC and Caspase-1 in liver tissue of rats with liver failure(P<0.05).Conclusion JDHY can inhibit the excessive activation of ROS-NLRP3 signaling pathway,reduce the release of pro-inflammatory factors,alleviate the level of liver inflammatory stress and antagonize liver failure.

关 键 词：解毒化瘀颗粒 ROS-NLRP3炎症小体信号通路 肝衰竭 

分 类 号：R285.5[医药卫生—中药学]