基于NLRP3/Caspase-1/GSDMD焦亡通路探讨涤浊化瘀方对非酒精性脂肪性肝炎小鼠的影响  

Mechanism of Dizhuo Huayu Decoction on nonalcoholic steatohepatitis mouse based on NLRP3/Caspase-1/GSDMD pyroptosis pathway

作　　者：尚东方[1,2] 赵文霞[2] 赵晨露 刘鸣昊[2] 刘晓彦[2] 王思颖 马素平[2] 朱平生[1] SHANG Dongfang;ZHAO Wenxia;ZHAO Chenlu;LIU Minghao;LIU Xiaoyan;WANG Siying;MA Suping;ZHU Pingsheng(Henan University of Chinese Medicine,Zhengzhou 450000,China;The First Affiliated Hospital of Henan University of CM,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学,郑州450000 [2]河南中医药大学第一附属医院,郑州450000

出　　处：《中华中医药杂志》2024年第11期6061-6065,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81904154);河南省中医学“双一流”创建科学研究专项(No.HSRP-DFCTCM-2023-3-13,No.HSRP-DFCTCM-2023-4-02);河南省科技攻关计划(No.242102311285);全国名中医传承工作室建设项目(No.国中医药办人教函〔2022〕245号)。

摘　　要：目的:观察涤浊化瘀方对非酒精性脂肪性肝炎(NASH)小鼠的干预效果,及对焦亡通路的影响。方法:将60只C57BL/6J小鼠随机分为6组,空白组,模型组,高、中、低剂量组和多烯组。空白组给予普通饲料,其余各组给予高脂胆碱缺乏饲料,造模的同时灌胃给药,时间12周。全自动生化分析仪检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)和总胆固醇(CHO)、甘油三酯(TG)水平;HE、油红O和Masson染色观察肝脏病理形态;RT-PCR和Western Blot检测肝组织NLRP3、Caspase-1、GSDMD mRNA和蛋白表达;ELISA检测肝组织白介素(IL)-18和IL-1β的表达。结果:与模型组比较,涤浊化瘀方可以改善肝脏脂质沉积、炎症细胞浸润和纤维化,涤浊化瘀方各剂量组均可以降低血清中ALT、AST、ALP和CHO的表达(P<0.01),高、中剂量组可以降低血清中TG的表达(P<0.01),各剂量组均可降低肝组织中NLRP3、GSDMD mRNA表达(P<0.05),高剂量组降低肝组织中Caspase-1 mRNA表达(P<0.05),各剂量组均可降低肝组织中NLRP3、Caspase-1、GSDMD蛋白的表达,高、中剂量组和多烯组可显著降低肝组织中IL-18和IL-1β表达(P<0.01)。结论:涤浊化瘀方可能通过抑制焦亡通路,减少炎症反应,发挥治疗NASH的作用。Objective:To observe the intervention effect of Dizhuo Huayu Decoction on non-alcoholic steatohepatitis(NASH)model in mice and its effect on pyroptosis pathway.Methods:Sixty C57BL/6J mice were randomly divided into six groups:blank group,model group,high,medium and low dose groups of the Dizhuo Huayu Decoction,and polyene phosphatidylcholine(PP)group.The blank group was given ordinary chow,and the other groups were given high-fat cholinedeficient chow.At the same time of modeling,the rats were given intragastric administration for 12 weeks.The levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total cholesterol(CHO)and triglyceride(TG)were detected by automatic biochemical analyzer.HE,oil red O and Masson staining were used to observe the pathological morphology of the liver.The mRNA and protein expressions of NLRP3,Caspase-1 and GSDMD in liver tissue were detected by RT-PCR and Western Blot.The expression of interleukin(IL)-18 and IL-1βin liver tissue was detected by ELISA.Results:Compared with the model group,Dizhuo Huayu Decoction could reduce liver lipid deposition,inflammatory cell infiltration and fibrosis,each dose group of Dizhuo Huayu Decoction could reduce the expression of ALT,AST,ALP and CHO in serum(P<0.01),the high and medium dose groups could reduce the expression of TG in serum(P<0.01),each dose group could reduce the expression of NLRP3 and GSDMD mRNA in liver tissue(P<0.05),the high dose group could reduce Caspase-1 mRNA expression in liver tissue(P<0.05),each dose group could reduce the protein expression of NLRP3,Caspase-1 and GSDMD in liver tissue and the high and medium dose groups and PP group could significantly reduce the expression of IL-18 and IL-1βin liver tissue(P<0.01).Conclusion:The Dizhuo Huayu Decoction may play a role in the treatment of NASH by inhibiting the pyroptosis pathway and reducing the inflammatory response.

关 键 词：非酒精性脂肪性肝炎 涤浊化瘀方 焦亡 NLRP3 CASPASE-1 GSDMD 

分 类 号：R285.5[医药卫生—中药学]