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

WU You; CHEN Yuan; HU Yaomu; SUN Boju; GUO Xiaoyuan; QIN Lingling; LIU Tonghua; WU Lili

doi:10.13422/j.cnki.syfjx.20230501

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

更新时间：2023-07-18

- Effect of Zishenwan on Intestinal Barrier and Transcriptome of Skeletal Muscle of db/db Diabetic Mice
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 16, Pages: 22-32(2023)
- 作者机构：
  
  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
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230501
  CLC： R2-0;R22;R285.5;R289;R33
- Published：20 August 2023，
  
  Published Online：13 February 2023，
  
  Received：10 October 2022，
- 稿件说明：
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吴悠,陈源,胡耀木等.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[J].中国实验方剂学杂志,2023,29(16):22-32.

WU You,CHEN Yuan,HU Yaomu,et al.Effect of Zishenwan on Intestinal Barrier and Transcriptome of Skeletal Muscle of db/db Diabetic Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(16):22-32.
吴悠,陈源,胡耀木等.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[J].中国实验方剂学杂志,2023,29(16):22-32. DOI： 10.13422/j.cnki.syfjx.20230501.

WU You,CHEN Yuan,HU Yaomu,et al.Effect of Zishenwan on Intestinal Barrier and Transcriptome of Skeletal Muscle of db/db Diabetic Mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(16):22-32. DOI： 10.13422/j.cnki.syfjx.20230501.

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（

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（

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（

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（

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。

Abstract

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 （

0.01）. Compared with the model group， Zishenwan significantly reduced fasting blood glucose， body weight， TC， TG， and HOMA-IR in db/db mice （

0.01）， 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 （

0.01）. Compared with the model group， Zishenwan improved the pathological changes in skeletal muscle and ileum， and increased the protein expression of occludin and ZO-1 in the ileum （

0.01）. Transcriptome analysis suggested that Zishenwan might improve skeletal muscle metabolism and increase insulin sensitivity in mice.

Conclusion

Zishenwan can improve glucose and lipid metabolism in db/db mice， and this effect may be related to its protection of intestinal barrier function and transcriptional regulation of skeletal muscle metabolism-related genes.

关键词

滋肾丸糖尿病骨骼肌肠道屏障转录组

Keywords

Zishenwandiabetesskeletal muscleintestinal barriertranscriptome

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