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
吴智鹏 1张荣臻 2王明刚 2张钰琴 3王秀峰2
作者信息
- 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
- 折叠
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,EUSA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、1L-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.
Abstract
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 divid-ed 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 model-ing.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-1β and IL-18 were measured by ELISA.ROS pro-duction was evaluated.The expression of NLRP3 and GSDMD in liver tissue was analyzed by immunohistochemistry.The expres-sion 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Key words
Jiedu Huayu Granules/ROS-NLRP3 inflammasome signaling pathway/Liver failure引用本文复制引用
基金项目
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)
广西中医药大学第一附属医院博士启动基金(2020BS003)
广西中医药大学桂派中医药传承创新团队项目(2022A001)
出版年
2024
NETL
NSTL
万方数据