摘要
目的基于TLR4/NF-κB/TNF-α信号通路探讨疏肝温胆汤对代谢综合征小鼠的作用及机制。方法将30只C57BL/6小鼠随机分为正常对照组(7只)、模型组(7只)、疏肝温胆汤组(8只,15.6 g·kg^(-1)·d^(-1))、阿托伐他汀钙组(8只,225 mg·kg^(-1)·d^(-1))。通过连续12周的高脂高果糖饲料喂养复制代谢综合征小鼠模型。造模4周后开始,疏肝温胆汤组及阿托伐他汀钙组按照上述剂量灌胃给药,每日1次,连续灌胃8周。测量小鼠体质量及体长,计算LEE’s指数;采用血糖仪检测小鼠空腹血糖(FPG);微板法检测血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)及血清游离脂肪酸(FFA)水平;ELISA法检测血清胰岛素(FINS)水平,计算胰岛素抵抗指数;HE及油红O染色法观察肝脏组织病理变化;Western Blot法检测肝脏组织Toll样受体4(TLR4)、核因子κB(NF-κB)及其磷酸化蛋白(p-NF-κB)、肿瘤坏死因子α(TNF-α)蛋白表达水平。结果与正常对照组比较,模型组小鼠的体质量、LEE’s指数、FPG、FINS、HOMA-IR、TC、TG、LDL-C、FFA水平均显著升高(P<0.05,P<0.01),HDL-C水平显著降低(P<0.01);肝脏组织显示肝细胞索排列紊乱,局部肿胀,部分肝细胞脂肪变性,胞质内有大量脂肪空泡及红色脂滴沉积,形态大小不一;肝脏组织TLR4、p-NF-κB/NF-κB、TNF-α蛋白表达水平显著升高(P<0.01)。与模型组比较,疏肝温胆汤组、阿托伐他汀钙组小鼠的体质量、LEE’s指数、FPG、FINS、HOMA-IR、TC、TG、LDL-C、FFA水平均显著降低(P<0.05,P<0.01),HDL-C水平明显升高(P<0.05);肝脏组织中脂滴空泡减少,肝索细胞排列及脂滴沉积情况均有所改善;肝脏组织TLR4、p-NF-κB/NF-κB、TNF-α蛋白表达水平均明显降低(P<0.05)。结论疏肝温胆汤能够改善代谢综合征小鼠的糖脂代谢及胰岛素抵抗状态,减轻肝脏脂质沉积及炎症反应,其作用机制可能与抑制TLR4/NF-κB/TNF-α信号通路有关。
Objective To investigate the effects and mechanisms of Shugan Wendan Decoction(SWD)on metabolic syndrome(MS)through the TLR4/NF-κB/TNF-αsignaling pathway.Methods Thirty C57BL/6 mice were randomly divided into four groups:normal control(n=7),model(n=7),SWD(n=8,15.6 g·kg^(-1)·d^(-1)),and Atorvastatin Calcium(n=8,225 mg·kg^(-1)·d^(-1)).A MS mouse model was established by feeding a high-fat and high-fructose diet for 12 weeks.After 4 weeks of modeling,SWD and Atorvastatin Calcium were administered intragastrically once daily for 8 weeks.Body mass and length were measured to calculate LEE’s index.Fasting plasma glucose(FPG)was detected using a glucometer.Serum total cholesterol(TC),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),and free fatty acids(FFA)levels were analyzed via microplate assays.Serum insulin(FINS)levels were measured by ELISA,and the homeostasis model assessment of insulin resistance(HOMA-IR)was calculated.Hepatic histopathology was observed using HE and Oil Red O staining.Western Blot was employed to quantify protein expression of Toll-like receptor 4(TLR4),nuclear factor-κB(NF-κB),phosphorylated NF-κB(p-NF-κB),and tumour necrosis factor-α(TNF-α)in liver tissues.Results Compared with the normal control group,the model group exhibited significant increases in body mass,LEE’s index,FPG,FINS,HOMA-IR,TC,TG,LDL-C,and FFA levels(P<0.05,P<0.01),with marked reductions in HDL-C(P<0.01).Hepatic tissues displayed disordered hepatocyte cord arrangement,localized swelling,steatosis,and abundant lipid vacuoles and red lipid droplets,varying in morphology and size.Protein expression of hepatic TLR4,p-NF-κB/NF-κB,and TNF-αwere significantly elevated(P<0.01).Compared with the model group,SWD and Atorvastatin Calcium groups showed decreased body mass,LEE’s index,FPG,FINS,HOMA-IR,TC,TG,LDL-C,and FFA levels(P<0.05,P<0.01),increased HDL-C(P<0.05),reduced lipid vacuoles,improved hepatic cord arrangement,and decreased protein expression of hepatic TLR4,p-NF-κB/NF-κB,and TNF-α(P<0.05).Conclusion SWD ameliorates glucose-lipid metabolism,insulin resistance,hepatic lipid deposition,and inflammation in MS mice,possibly via suppressing the TLR4/NF-κB/TNF-αsignaling pathway.
作者
彭伟航
张为章
黄小燕
于丽媛
蔡海荣
庄杰钦
赵帅
陈伯钧
PENG Weihang;ZHANG Weizhang;HUANG Xiaoyan;YU Liyuan;CAI Hairong;ZHUANG Jieqin;ZHAO Shuai;CHEN Bojun(The Second School of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China)
出处
《中药新药与临床药理》
北大核心
2025年第8期1243-1248,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
广东省中医药局科研项目(20233021)
中医湿证国家重点实验室建设专项(20225008)
广州市科技局市校(院)联合资助项目(202201020352)
广东省中医院博士后专项科研课题(10814)。
