滋肾丸对db/db糖尿病模型小鼠肠道屏障功能及骨骼肌转录组的影响  

Effect of Zishenwan on Intestinal Barrier and Transcriptome of Skeletal Muscle of db/db Diabetic Mice

作　　者：吴悠 陈源 胡耀木 孙伯菊 郭晓媛[3] 秦灵灵[1] 刘铜华[1] 吴丽丽[1] WU You;CHEN Yuan;HU Yaomu;SUN Boju;GUO Xiaoyuan;QIN Lingling;LIU Tonghua;WU Lili(Key Laboratory of Health Cultivation of the Ministry of Education,Key Laboratory of Health Cultivation of Beijing,Beijing University of Chinese Medicine,Beijing 100029,China;School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China;Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;Clinical College of Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730101,China;Fengtai District Hospital of Chinese Medicine,Beijing 100076,China)

机构地区：[1]北京中医药大学教育部中医养生学重点实验室,中医养生学北京市重点实验室,北京100029 [2]北京中医药大学生命科学学院,北京102488 [3]北京中医药大学东方医院,北京100078 [4]甘肃中医药大学中医临床学院,兰州730101 [5]丰台区中医医院,北京100076

出　　处：《中国实验方剂学杂志》2023年第16期22-32,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：高等学校学科创新引智计划项目(B20055);科技部国家国际合作基地项目(2015B01022)。

摘　　要：目的:探索滋肾丸对自发型糖尿病模型db/db小鼠糖脂代谢的作用,并基于肠道屏障功能和骨骼肌转录组测序结果,探索其在体内改善糖尿病作用的可能机制。方法:使用液相色谱-质谱联用技术(LC-MS/MS)对滋肾丸进行成分分析。将6周龄db/db小鼠16只分为模型组、滋肾丸组,将野生型小鼠8只作为正常组。滋肾丸灌胃给药共6周,期间测量小鼠空腹血糖、体质量、进食量。检测小鼠血清总胆固醇(TC)及甘油三酯(TG)水平,检测小鼠空腹胰岛素水平并计算胰岛素抵抗指数(HOMA-IR)。给药结束后,取小鼠骨骼肌、回肠组织,行苏木素-伊红(HE)染色,同时使用免疫组化法检测回肠紧密连接蛋白闭合蛋白(Occludin)和闭锁连接蛋白-1(ZO-1)的表达。使用转录组学测序检测小鼠骨骼肌转录本,并对差异表达基因进行富集分析。结果:从滋肾丸中识别出多种中药活性成分。与正常组比较,模型组小鼠空腹血糖、体质量、TC、TG和HOMA-IR显著升高(P<0.01);与模型组比较,滋肾丸给药显著降低db/db小鼠的空腹血糖、体质量、TC、TG、HOMA-IR(P<0.01),而对进食量差异无统计学意义。与正常组比较,模型组小鼠的骨骼肌出现脂质沉积,同时回肠结构改变,肠道Occludin和ZO-1的蛋白表达水平显著降低(P<0.01);与模型组比较,滋肾丸改善了小鼠骨骼肌和回肠的病理改变,显著升高回肠Occludin和ZO-1的蛋白表达(P<0.01)。转录组提示,滋肾丸可能改善了小鼠骨骼肌的代谢,并增加了胰岛素敏感性。结论:滋肾丸可以改善db/db小鼠的糖脂代谢,此作用可能和其对肠道屏障功能的保护和对骨骼肌代谢相关基因的转录调控有关。Objective:To explore the effect of Zishenwan on glucose and lipid metabolism in spontaneous type 2 diabetes(db/db)mice and investigate the underlying mechanism for improving diabetes based on intestinal barrier function and skeletal muscle transcriptome sequencing results.Method:Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was used to analyze the components of Zishenwan.Sixteen 6-week-old db/db mice were divided into a model group and a Zishenwan group,while eight wild-type mice were assigned to the normal group.The Zishenwan group received oral administration of drugs for six weeks,during which fasting blood glucose,body weight,and food intake were measured.Serum total cholesterol(TC)and triglyceride(TG)levels were determined,and fasting insulin levels were measured to calculate the homeostatic model assessment of insulin resistance(HOMA-IR).After the treatment,skeletal muscle and ileum tissues were collected,followed by hematoxylin-eosin(HE)staining.Immunohistochemistry was used to detect the expression of tight junction proteins occludin and zonula occludens-1(ZO-1)in the ileum.Transcriptome sequencing was performed to detect the skeletal muscle transcriptome,and enrichment analysis was conducted for differentially expressed genes.Result:Multiple active components were identified in Zishenwan.Compared with the normal group,the model group showed increased fasting blood glucose,body weight,TC,TG,and HOMA-IR(P0.01).Compared with the model group,Zishenwan significantly reduced fasting blood glucose,body weight,TC,TG,and HOMA-IR in db/db mice(P0.01),while there was no statistically significant difference in food intake.Compared with the normal group,the model group exhibited lipid deposition in skeletal muscle,as well as structural changes in the ileum,with significant decreases in the protein expression levels of intestinal occludin and ZO-1(P0.01).Compared with the model group,Zishenwan improved the pathological changes in skeletal muscle and ileum,and increased the protein expression of occludin and

关 键 词：滋肾丸 糖尿病 骨骼肌 肠道屏障 转录组 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33