四川黑茶药膳配方通过重塑肠道菌群和短链脂肪酸代谢改善肥胖小鼠肾脏脂质紊乱  

Sichuan Dark Tea-Based Medicated Dietary Formula Improves Obesity-Induced Renal Lipid Metabolism Disorder in Mice by Remodeling Gut Microbiota and Short-Chain Fatty Acid Metabolism

作　　者：李卉 张历涵 黄蓉双 任倩[1] 郭帆 石敏[1] 杨乐天 于洋[1] 马良[1] 付平[1] LI Hui;ZHANG Lihan;HUANG Rongshuang;REN Qian;GUO Fan;SHI Min;YANG Letian;YU Yang;MA Liang;FU Ping(Kidney Research Institute,Department of Nephrology,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院肾脏内科肾脏病研究所,成都610041

出　　处：《四川大学学报（医学版）》2023年第6期1112-1120,共9页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.82070711);四川省科技厅重点研发计划(No.2022YFS0589、No.2022YFS0325);四川省科技创新苗子工程项目(No.MZGC20230017)资助。

摘　　要：目的 探讨以四川黑茶为基础的药膳配方(元清)对高脂饮食诱导的肥胖小鼠的肾脏保护作用并探究其具体机制。方法 将雄性C57BL/6小鼠随机分为对照组、模型组和治疗组,每组8只。对照组饲喂普通维持饲料和纯净水,另外两组给予高脂饲料喂养12周以建立肥胖模型。此后模型组继续饲喂高脂饲料,治疗组同时给予元清12周,期间每周记录小鼠体质量。12周后处死小鼠,取血清检测三酰甘油(TG)、总胆固醇(TC)、天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)及白蛋白水平以评估肝功能,提取肾脏脂质检测肾脏TG及TC含量,过碘酸-雪夫(Periodic Acid-Schiff, PAS)和油红O染色评估肾脏病理损伤。Western blot检测肾脏组织中的磷酸化AMPK (pAMPK)/AMPK比值。PCR及Western blot检测肾脏组织中调控脂肪酸氧化蛋白乙酰辅酶a羧化酶1(acetyl-CoA carboxylase 1, ACC1)、肉毒碱酰基转移酶1 (carnitine acyltransferase 1, CTP1)、过氧化物酶体增殖物激活受体γ(peroxisome proliferators-activated receptor γ, PPARγ)、过氧化物酶体增殖物激活受体γ辅助激活因子-1α(peroxisome proliferator-activated receptor gamma coactivator-1 alpha, PGC1α)及脂肪酸合成关键分子胆固醇调节元件结合蛋白-1(sterol-regulatory element binding proteins, SREBP-1)、脂肪酸合成酶(fatty acid synthase, FASN)、硬脂酰辅酶a去饱和酶1 (stearoyl-CoA desaturase 1, SCD1)的表达水平。16SrRNA及代谢组学分析其肠道内容物中肠道菌群及其代谢产物。结果 与对照组相比,模型组小鼠肝质量(P=0.000 3),血清ALT(P<0.000 1)、AST(P=0.000 1)水平,肾脏TC (P=0.019 1)、TG(P=0.010 1)水平升高,肾脏脂质沉积。与模型组相比,治疗组有效降低小鼠肝质量(P=0.031 6),改善血清AST (P=0.001 2)、ALT (P=0.002 7),肾脏TC (P=0.020 0)、TG (P=0.049 9)异常水平,同时显著改善肾脏脂质沉积。治疗组与模型组相比,pAMPK/AMPK比值升高。与对照组相比,模型组小鼠肾脏Objective To investigate the renoprotective effects of a Sichuan dark tea-based medicated dietary formula(alternatively referred to as Qing,or clarity in Chinese)on mice with diet-induced obesity(DIO)and to explore the specific mechanisms involved.Methods Male C57BL/6 mice were randomly assigned to three groups,a control group,a DIO group,and a Qing treatment group,or the Qing group,with 8 mice in each group.The mice in the control group were given normal maintenance feed and purified water,and the other two groups were fed a high-fat diet for 12 weeks to establish the DIO model.After that,high-fat diet continued in the DIO group,while the Qing group was given Qing at the same time for 12 weeks,during which period the weight of the mice was monitored and recorded every week.The mice were sacrificed after 12 weeks.Serum samples were collected and the levels of triglyceride(TG),total cholesterol(TC),alanine aminotransferase(ALT),aspartate aminotransferase(AST),and albumin were measured to evaluate liver function.In addition,renal lipids were extracted to determine the levels of TG and TC in the kidney and periodic acid-Schiff(PAS)and oil red O stainings were performed to evaluate kidney pathological injury.Western blot was performed to determine the phosphorylated AMPK(pAMPK)/AMPK ratio in the kidney tissue.RT-qPCR and Western blot were used to determine the expression of proteins related to fatty acid oxidation,including acetyl-CoA carboxylase 1(ACC1),carnitine acyltransferase 1(CTP1),peroxisome proliferators-activated receptorγ(PPARγ),peroxisome proliferators-activated receptor-1α(PPAR1α),sterol-regulatory element binding proteins(SREBP-1),and key proteins related to lipid synthesis,including fatty acid synthase(FASN)and stearoyl-coenzyme A desaturase 1(stearoyl-CoA desaturase)in the kidney tissue.16SrRNA and metabolomics were applied to analyze the gut microbiota in the intestinal contents and its metabolites.Results Compared with those of the control group,the levels of liver mass(P=0.0003),serum ALT(P<0.00

关 键 词：高脂饮食 肥胖 肾脏脂质代谢 慢性肾脏病 四川黑茶 

分 类 号：TS272.5[农业科学—茶叶生产加工]