泽泻汤通过调控NLRP3/Caspase-1/GSDMD焦亡途径减缓大鼠非酒精性脂肪肝的机制研究

Study on Mechanism of Slowing Down Non-alcoholic Fatty Liver Disease in Rats by Regulating NLRP3/Caspase-1/GSDMD Pyrosis Pathway by Zexie Decoction(泽泻汤)

目的 探究泽泻汤减缓大鼠非酒精性脂肪肝(non-alcoholic fatty liver disease, NAFLD)的机制,揭示(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)/半胱天冬酶1(cysteinyl aspartate specific proteinase-1,Caspase-1)/消皮素-D(gasdermin-D,GSDMD)焦亡途径在其过程中的调控作用。方法 采用高脂饲料喂养复制NAFLD大鼠模型,将SD大鼠随机分为对照组、模型组、泽泻汤正常剂量组、泽泻汤高剂量组,造模成功后,分别给予对应剂量泽泻汤及等体积生理盐水灌胃给药30 d,全自动生物化学分析仪检测各组大鼠血清甘油三酯(triglyceride, TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol, LDL-C)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol, HDL-C)、丙氨酸氨基转移酶(alanine transaminase, ALT)、天冬氨酸氨基转移酶(aspartate transaminase, AST),HE和油红O染色观察大鼠肝脏形态学变化,RT-qPCR和Wes全自动蛋白质印迹定量分析系统检测大鼠肝脏NLRP3、GSDMD、Caspase-1、白介素-1 β(interleukin-1β,IL-1 β)基因mRNA及蛋白表达。结果 与对照组相比,模型组大鼠血清TG、TC、LDL、ALT、AST表达水平显著升高(P<0.01),HDL-C表达水平显著降低(P<0.01),肝脏结构显著异常,大鼠肝脏NLRP3、GSDMD、Caspase-1、IL-1β基因mRNA及蛋白表达显著升高(P<0.05或P<0.01),给予不同剂量的泽泻汤干预后,NAFLD大鼠血脂水平明显改善,ALT、AST表达水平显著降低(P<0.05或P<0.01),肝脏结构异常得以改善,其中泽泻汤高剂量为实验最佳治疗剂量,泽泻汤高剂量治疗后,NAFLD大鼠肝脏GSDMD、IL-1β基因mRNA及蛋白表达显著降低(P<0.05或P<0.01),NLRP3、Caspase-1基因mRNA表达显著降低(P<0.05或P<0.01)。结论 泽泻汤可能通过调控NLRP3/Caspase-1/GSDMD焦亡途径减缓大鼠非酒精性脂肪肝进程。

Objective To investigate the mechanism of Zexie Decoction(泽泻汤)in slowing down nonalcoholic fatty liver dis-ease(NAFLD)in rats and reveal the regulatory role of Nucleotide binding oligomerization domain like receptor protein 3(NLRP3)/Caspase-1/Gasdermin-D(GSDMD)pyroptosis pathway in this process.s Methods The SD rats were randomly divid-ed into control group,model group,normal dose group and high dose group of Zexie Decoction by feeding high-fat feed.After successful modeling,the corresponding dose of Zexie Decoction and the equal volume of normal saline were given for 30 days.The automatic biochemical analyzer was used to detect the serum levels of triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in each group.The morphological changes of the liver of rats were observed by HE and oil red O stai-ning,and RT-qPCR and Wes automatic western blot quantitative analysis system were used to detect the expressions of NLRP3,GSDMD,Caspase-1,interleukin-1β(IL-1β)gene mRNA and protein in rat liver.s Results Compared with those of the control group,the serum levels of of TG,TC,LDL,ALT and AST in the model group increased significantly(P<0.01)and HDL-C ex-pression levels decreased significantly(P<0.01).The liver structure was significantly abnormal and the expression of gene mR-NA and protein of NLRP3,GSDMD,Caspase-1 and IL-1βin the liver of the rats were significantly increased(P<0.05 or P<0.01).After different doses of Zexie Decoction intervention,the blood lipid level of NAFLD rats was significantly improved,and the expression levels of ALT and AST were significantly reduced(P<0.05 or P<0.01).The structural abnormalities of the liver were improved,and the high dose of Zexie Decoction was the best therapeutic dose in this experiment.After high-dose treat-ment,the expressions of GSDMD,IL-1βgene mRNA and protein in NAFLD rats was significantly reduced(P<0.05 or P<0.01),and the expressions of mRNA in NLRP3 and Caspase-1 genes were significantly reduced(P<0.05 or P<0.01).Con-clusion Con-clusion Zexie Decoction may slow down the process of NAFLD rats by regulating the NLRP3/Caspase-1/GSDMD pyrosis path-way.