去脂软肝方对NASH大鼠NLRP3炎症小体及相关因子表达的影响

Effects of Quzhi Ruangan Formula on the expression of NLRP3 infammasome and related factors in rats with non-alcoholic steatohepatitis

目的:观察去脂软肝方对非酒精性脂肪性肝炎(NASH)的防治作用及对NLRP3炎症小体的影响。方法:36只大鼠随机分为正常组,模型组,去脂软肝方低、中、高剂量组和辛伐他汀组,每组6只。正常组用普通饲料饲养,其余各组给予高脂饲料饲养14周复制NASH大鼠模型,造模的同时予相应的药物进行干预。全自动生化仪检测大鼠血清中ALT、AST、TC、TG、LDL-C、HDL-C含量,GPO-PAP法检测肝脏TC、TG含量,HE染色及油红O染色镜下观察大鼠肝组织病理改变,实时荧光定量PCR和免疫组化二步法分别检测NLRP3、Caspase-1 mRNA和蛋白表达水平,ELISA检测肝脏中IL-1β、IL-18含量。结果:与模型组比较,去脂软肝方干预后大鼠肝组织的脂质沉积和炎症细胞浸润程度减轻,中、高剂量组血清ALT、AST、TC、TG、LDL-C及肝脏TC、TG含量均显著下降(P<0.01,P<0.05),各剂量组HDL-C含量显著升高(P<0.01),中、高剂量组NLRP3、Caspase-1 mRNA及蛋白表达水平均显著下降(P<0.01,P<0.05),中、高剂量组肝脏IL-1β和IL-18表达水平均显著降低(P<0.01)。结论:去脂软肝方可下调NASH模型大鼠NLRP3炎症小体及下游炎症因子的表达,减轻肝脏脂肪变性和炎症反应。

Objective:To explore the effects of Quzhi Ruangan Formula(QZRGF)on non-alcoholic steatohepatitis(NASH)and the effects of QZRGF on the expression of NLRP3.Methods:Thirty-six SD rats were randomly divided into the control group,the model group,the QZRGF low-dose group,the QZRGF medium-dose group,the QZRGF high-dose group,and the simvastatin group,6 rats in each group.The rats in control group was fed with normal diet,and the other groups were fed with high-fat diet for 14 weeks to replicate the NASH rat model.During the model replication,rats in each group were intervened with related drugs.Serum levels of liver function indexes(ALT,AST),blood lipid indexes(TC,TG,LDL-C,HDL-C)and liver lipid indexes(TC,TG)were detected in all groups by automatic biochemical analyzer and GPO-PAP method.HE staining and oil red O staining were used to observe pathological changes of liver.RT-qPCR was used to detect the expression levels of NLRP3 mRNA and Caspase-1 mRNA.Immunohistochemistry technique was used to detect the protein expression of NLRP3 and Caspase-1.ELISA was used to detect the concentrations of IL-1βand IL-18 in the liver tissue.Results:Compared with the model group,QZRGF treatment alleviated the degree of fat vacuoles,loose cytoplasm and inflammatory cell infiltration in rat liver,the mediumdose and high-dose groups decreased the contents of ALT,AST,TC,TG and LDL-C in serum and TG,TC in liver tissae(P<0.01,P<0.05),three doses groups increased the contents of HDL-C(P<0.01).Furthermore,the medium-dose and high-dose groups also down-regulated the mRNA and protein expression of NLRP3 and Caspase-1(P<0.01,P<0.05),as well as the concentrations of IL-1βand IL-18(P<0.01).Conclusion:QZRGF could alleviate hepatic steatosis and inflammation,which may be achieved by downregulating the expression of NLRP3 inflammasome and downstream inflammatory factors in NASH model rats.