酪酸梭菌对db/db小鼠肾组织的保护作用及机制  

Protective effect of Clostridium butyricum on renal tissue in db/db mice and its mechanism

作　　者：黄诗琴 胡婷 叶凯丽 赵艳玲[1] Huang Shiqin;Hu Ting;Ye Kaili;Zhao Yanling(Department of Nephrology,the Second Affiliated Hospital of Wenzhou Medical University,Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第二医院肾脏内科,温州325000

出　　处：《中华内分泌代谢杂志》2023年第6期506-516,共11页Chinese Journal of Endocrinology and Metabolism

基　　金：浙江省公益技术应用研究资助课题 (LGD19H070003)。

摘　　要：目的研究酪酸梭菌对db/db小鼠肾组织的保护作用及机制。方法14周龄db/db小鼠按数字表法随机分为糖尿病肾病组(db/db组,n=10)和酪酸梭菌治疗组(db/db+Cb组,n=7),选用同周龄db/m小鼠为正常对照组(db/m组,n=10)。db/m和db/db小鼠给予0.9%氯化钠溶液灌胃,db/db+Cb小鼠给予等量的酪酸梭菌溶液灌胃,连续灌胃8周。检测血肌酐、空腹血糖和尿白蛋白/肌酐比值(ACR)等指标;HE染色观察肾组织病理变化;实时荧光定量PCR检测肾组织过氧化物酶体增殖物活化受体γ协同刺激因子-1α(PGC-1α)mRNA表达情况;免疫组化及蛋白免疫印迹法测定肾组织核因子-κB(NF-κB)、胰升糖素样肽1受体(GLP-1R)、腺苷酸活化蛋白激酶(AMPK)蛋白的表达。采用16S rRNA法、液相色谱质谱联用和气相色谱质谱联用分别测定肠道菌群、血清和粪便短链脂肪酸(SCFAs)水平。结果与db/db小鼠相比,db/db+Cb小鼠通过补充酪酸梭菌后一般状态改善,空腹血糖、血尿素氮、ACR、血肌酐、肾脏组织中白细胞介素6(IL-6)水平降低(均P<0.05);病理显示,小鼠肾组织损伤有不同程度的改善;肾组织中PGC-1αmRNA表达量上升(P<0.05);肾组织NF-κB蛋白表达下降,GLP-1R、磷酸化腺苷酸活化激酶(p-AMPK)/AMPK蛋白表达上调(均P<0.05);酪酸梭菌调节了肠道微生物群的组成,血清和粪便中总SCFAs含量升高(均P<0.05)。结论酪酸梭菌能增加肾组织GLP-1R表达,促进AMPK磷酸化,减轻小鼠肾组织损伤,推测与调节富产SCFAs菌群有关。Objective To investigate the effect of Clostridium butyricum on renal tissue of db/db mice and to explore its mechanism.Methods Fourteen-week-old db/db mice were divided into db/db group(n=10)and db/db+Cb group(n=7)according to random number table method.Age-matched db/m mice were selected as the normal control group.The db/m and db/db mice were administered 0.9%sodium chloride solution by gavage,while the db/db+Cb mice were administered an equivalent amount of Clostridium butyricum solution by gavage for 8 weeks.Serum creatinine,fasting blood glucose,urinary albumin to creatinine ratio(ACR)and other biochemical indicators were also detected.HE staining was used to observe the pathological changes of kidney tissue.The expressions of peroxisome proliferators-activated receptorγcoactivator-1α(PGC-1α)mRNA were detected by realtime PCR,while the expressions of nuclear factor-κB(NF-κB),glucagon-like peptide 1 receptor(GLP-1R),and adenosine monophosphate-activated protein kinase(AMPK)in kidney tissue were determined by immunohistochemistry and Western blotting.The levels of intestinal flora,serum and fecal short-chain fatty acids(SCFAs)were measured by 16S rRNA,liquid chromatograph-mass spectrometer,and gas chromatograohy-mass spectrometry respectively.Results Compared to db/db mice,db/db+Cb mice showed improvement in general condition after supplementation with Clostridium butyricum.Fasting blood glucose,blood urea nitrogen,albumin-to-creatinine ratio(ACR),blood creatinine,and levels of interleukin-6(IL-6)in kidney tissue were reduced(all P<0.05).The pathology showed various degrees of amelioration of kidney tissue injury in mice.The expression of PGC-1αmRNA increased in kidney tissue(P<0.05).Decreased expression of NF-κB protein,as well as increased expression of GLP-1R and phosphorylated(p-)AMPK/AMPK protein(all P<0.05)were detected in kidney tissues.Clostridium butyricum modulated the composition of the gut microbiota with elevated total SCFAs in blood and feces.Conclusion Clostridium butyricum increased the

关 键 词：糖尿病肾病 酪酸梭菌 肠道菌群 短链脂肪酸 

分 类 号：R692.9[医药卫生—泌尿科学] R587.2[医药卫生—外科学]