芍药苷抑制NLRP3炎症小体通路对db/db小鼠肝脏炎症和纤维化的影响  

Effects of paeoniflorin on inflammation and fibrosis in db/db micevia regulating NLRP3 inflammasome pathway

作　　者：王安丽 朱启金 吴永贵[2] 夏玲玲[1] Wang Anli;Zhu Qijin;Wu Yonggui;Xia Lingling(Dept of Infectious Diseases,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Nephrology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学第一附属医院感染病科,合肥230022 [2]安徽医科大学第一附属医院肾脏内科,合肥230022

出　　处：《安徽医科大学学报》2022年第11期1687-1693,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81904035)。

摘　　要：目的探讨芍药苷(PF)对NLRP3炎症小体通路介导的db/db小鼠肝脏损伤的影响。方法以db/db小鼠作为2型糖尿病小鼠模型,db/m小鼠作为正常对照组。小鼠分为6组:正常对照(db/m)组、正常对照+PF(PF)组、糖尿病(db/db)组、糖尿病+PF 25 mg/kg(db/db+PF 25 mg/kg)组、糖尿病+PF 50 mg/kg(db/db+PF 50 mg/kg)组、糖尿病+PF 100 mg/kg(db/db+PF 100 mg/kg)组。PF灌胃12周后,收集小鼠血清样本用于检测丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆固醇(TC)、三酰甘油(TG)和游离脂肪酸(FFA)水平。对小鼠肝脏组织进行HE染色、油红O染色和天狼星红染色观察肝脏病理损伤程度。免疫组化法检测肝脏组织F4/80、α-SMA和ColⅢ蛋白表达水平。Western blot法检测肝脏组织白介素(IL)-1β、IL-18、肿瘤坏死因子-α(TNF-α)、NOD样受体蛋白-3(NLRP3)、凋亡相关点样蛋白(ASC)和半胱天冬酶-1(Caspase-1)蛋白表达水平。结果与db/m组相比,db/db组血清ALT、AST、TC、TG和FFA水平升高,PF灌胃后这些指标降低。肝脏组织病理学检查显示,与db/m组相比,PF组肝脏病理无明显改变,db/db组小鼠肝脏组织脂质堆积、炎症细胞浸润和胶原沉积增加,PF处理减少了肝脏脂质堆积、炎症细胞浸润和胶原沉积。与db/m组相比,db/db组小鼠肝脏促炎细胞因子(IL-1β、IL-18和TNF-α)、F4/80、α-SMA、ColⅢ、NLRP3、ASC和Caspase-1蛋白表达升高,PF灌胃后这些蛋白均下降。结论PF抑制NLRP3炎症小体通路,减轻了db/db小鼠肝脏的炎症反应和纤维化。Objective To investigate the effect of paeoniflorin(PF)on liver injury mediated by NLRP3 inflammasome pathway in db/db mice.Methods In this study,db/db mice were used as the mice model of type 2 diabetic while db/m mice were used as control group.The mice were randomly divided into six groups:db/m group,db/m+PF group,db/db group,db/db+PF 25 mg/kg group,db/db+PF 50 mg/kg group,db/db+PF 100 mg/kg group.After 12 weeks of PF gavage,mouse serum samples were collected to detect the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG)and free fat acid(FFA).HE staining,oil red O staining and Sirius red staining were used to observe the degree of pathological injury of liver.The expression of F4/80,α-SMA and ColⅢprotein was detected by immunohistochemistry.The expression of interleukin-1β(IL-1β),interleukin-18(IL-18),tumor necrosis factor-α(TNF-α),NOD-like receptor 3(NLRP3),apotpsis associated spck-like protein(ASC)and cysteinyl asparate specific proteinase-1(Caspase-1)was detected by Western blot.Results Compared with db/m group,the levels of serum ALT,AST,TC,TG and FFA increased in db/db group,and these indexes decreased after PF gavage.Compared with db/m group,histopathological examination of the liver revealed increased hepatic tissue lipid accumulation,inflammatory cell infiltration,and collagen deposition in db/db mice,and PF treatment reduced hepatic lipid accumulation,inflammatory cell infiltration,and collagen deposition.Meanwhile,compared with db/m group,the expression of proinflammatory cytokines(IL-1β,IL-18 and TNF-α),F4/80,α-SMA,ColⅢ,NLRP3,ASC and Caspase-1 protein in the liver of db/db mice increased,and the expression of these proteins decreased after gavage with PF.Conclusion PF inhibited the NLRP3 inflammatory pathway and attenuated liver inflammation and fibrosis in db/db mice.

关 键 词：芍药苷 糖尿病性肝损伤 炎症 纤维化 NLRP3炎症小体通路 

分 类 号：R587.29[医药卫生—内分泌]