达格列净对代谢相关脂肪性肝病小鼠模型肠道菌群的影响及其机制分析

郑彩云; 俞丽丽; 田晓旭; 戴亨纷

doi:10.12449/JCH251116

达格列净对代谢相关脂肪性肝病小鼠模型肠道菌群的影响及其机制分析

DOI: 10.12449/JCH251116

1.
福建医科大学附属福清市医院临床药学室，福州 350300
2.
福建医科大学附属福清市医院科教科，福州 350300
3.
福建医科大学附属福州市第一总医院药学部，福州 350009

基金项目:

福建省自然科学基金 (2025J011339);

福州市卫生健康科技计划 (2022-S-wq1);

福建医科大学启航基金 (2020QH1346)

伦理学声明：本研究方案于2024年8月18日经福建医科大学实验动物伦理委员会审批，批号：IACUCFJMU 2024-Y-2092，符合实验动物管理与使用准则。

利益冲突声明：本文不存在任何利益冲突。

作者贡献声明：郑彩云负责设计课题和论文框架，起草论文；郑彩云和俞丽丽负责实验操作，研究过程的实施；俞丽丽和田晓旭负责数据收集，统计学分析；郑彩云和戴亨纷负责论文修改和绘制图表；戴亨纷负责拟定写作思路，指导撰写文章并最后定稿。

详细信息

通信作者:
戴亨纷， hengfendai2011@163.com （ORCID： 0000-0002-2037-4208）

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出版历程
- 收稿日期: 2025-06-16
- 录用日期: 2025-09-05
- 出版日期: 2025-11-25

Effect and mechanism of dapagliflozin on gut microbiota in a mouse model of metabolic associated fatty liver disease

1.
Department of Clinical Pharmacy，Fuqing Hospital Affiliated to Fujian Medical University，Fuzhou 350300，China
2.
Department of Science and Education，Fuqing Hospital Affiliated to Fujian Medical University，Fuzhou 350300，China
3.
Department of Pharmacy，Fuzhou First General Hospital Affiliated with Fujian Medical University，Fuzhou 350009，China

Research funding:

Natural Science Foundation of Fujian Province (2025J011339);

Fuzhou Health Science and Technology Project (2022-S-wq1);

Startup Fund of Scientific Research, Fujian Medical University (2020QH1346)

More Information

Corresponding author: DAI Hengfen， hengfendai2011@163.com （ORCID： 0000-0002-2037-4208）

摘要

摘要: 目的探讨达格列净对代谢相关脂肪性肝病（MAFLD）小鼠肝脂代谢及肠道微生态的影响，并阐明其潜在作用机制。方法 50只C57雄性小鼠随机分为正常组（Control组）、2型糖尿病合并代谢相关脂肪性肝病组（MAFLD组）、达格列净组（DAPA组）、米屈肼组（THP组）、达格列净联合米屈肼组（DAPA+THP组），每组各10只。采用高脂饮食联合链脲佐菌素诱导MAFLD小鼠模型。通过组织病理学、血糖血脂等生化指标评估疗效，并利用转录组学和宏基因组学分析差异基因及菌群变化。正态分布的计量资料多组间比较采用单因素方差分析，两两比较采用LSD检验；非正态分布的计量资料多组间比较采用Kruskal-Wallis H检验，两两比较采用Nemenyi检验。结果组织病理学结果显示，MAFLD组小鼠表现为过量的脂质沉积，肝细胞出现脂肪样变；与MAFLD组相比，DAPA组肝细胞脂肪样变显著改善，THP组、DAPA+THP组改善效果无DAPA组明显。与Control组相比，MAFLD组空腹血糖显著升高（P<0.05），血清ALT、AST、MDA、TC、TG、LDL-C均显著升高（P值均<0.05），HDL-C明显降低（P<0.05）。与MAFLD组相比，DAPA组、THP组和DAPA+THP组血清ALT、AST显著降低（P值均<0.05）。16S rRNA测序显示，MAFLD组较Control组，小鼠肠道菌群发生显著改变，MAFLD组的Firmicutes、Lactobacillaceae明显增加，Bacteroidetes、S24-7和Erysipelotrichaceae明显减少。DAPA组、THP组和DAPA+THP组可调节上述菌群趋向正常水平。肝脏转录组学分析主要富集的代谢通路包括类固醇激素生物合成、胆汁分泌、炎性介质调节TRP（瞬时受体电位通道）、脂肪酸延伸、脂质生物代谢过程等，关联的基因主要涉及脂质代谢关键靶点Acot2、Angptl4、Scd2和Npc1l1。结论 DAPA可能通过类固醇激素生物合成、胆汁分泌、炎性介质调节TRP、脂肪酸延伸通路改善MAFLD，调节肠道菌群稳态。
- 代谢相关脂肪性肝病 /
- 胃肠道微生物组 /
- 转录组 /
- 达格列净 /
- 米屈肼
Abstract: Objective To investigate the effect of dapagliflozin on liver lipid metabolism and gut microecology in mice with metabolic associated fatty liver disease （MAFLD）， and to clarify its potential mechanism. Methods A total of 50 male C57 mice were randomly divided into Control group， type 2 diabetes+MAFLD group （MAFLD group）， dapagliflozin group （DAPA group）， meldonium group （THP group）， and dapagliflozin+meldonium group （DAPA+THP group）， with 10 mice in each group. High-fat diet combined with streptozotocin was used to establish a mouse model of MAFLD. Treatment outcomes were assessed based on histopathology and biochemical parameters such as blood glucose and blood lipid levels， and the transcriptomic and metagenomic analyses were used to identify differentially expressed genes and the changes in gut microbiota. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups， and the least significant difference t-test was used for comparison between two groups； the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups， and the Nemenyi test was used for comparison between two groups. Results Histopathological examination showed that the mice in the MAFLD group had excessive lipid deposition and hepatocyte steatosis； compared with the MAFLD group， the DAPA group had a significant improvement in hepatocyte steatosis， while the THP group and the DAPA+THP group had a less significant improvement compared with the DAPA group. Compared with the Control group， the MAFLD group had a significant increase in fasting blood glucose （P<0.05）， significant increases in the serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， malondialdehyde， total cholesterol， triglyceride， and low-density lipoprotein cholesterol （P<0.05）， and a significant reduction in high-density lipoprotein cholesterol （P<0.05）. Compared with the MAFLD group， the DAPA group， the THP group， and the DAPA+THP group had significant reductions in the serum levels of ALT and AST （P<0.05）. The results of 16S rRNA sequencing showed that compared with the Control group， the MAFLD group had significant changes in gut microbiota， with an increase in Firmicutes and a reduction in Bacteroidetes， as well as reductions in S24-7 and Erysipelotrichaceae and an increase in Lactobacillaceae. The levels of the above flora were upregulated to normal levels in the DAPA group， the THP group， and the DAPA+THP group. The liver transcriptomic analysis showed that the enriched metabolic pathways included steroid hormone biosynthesis， bile secretion， inflammatory mediator regulation of TRP， fatty acid elongation， and lipid biodegradation processes， and the related genes mainly involved the key targets of lipid metabolism such as Acot2， Angptl4， Scd2， and Npc1l1. Conclusion Dapagliflozin can alleviate MAFLD through the pathways such as steroid hormone biosynthesis， bile secretion， inflammatory mediator regulation of TRP， and fatty acid elongation， as well as by regulating gut microbiota homeostasis.
- Metabolic Associated Fatty Liver Disease /
- Gastrointestinal Microbiome /
- Transcriptome /
- Dapagliflozin /
- Meldonium

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注： a，各组小鼠空腹血糖动态变化；b，链脲佐菌素注射前糖耐量实验；c，链脲佐菌素注射后糖耐量实验。

图 1 小鼠血糖的变化

Figure 1. Changes of blood glucose of mice

下载: 全尺寸图片幻灯片

图 2 小鼠肝组织HE染色及油红O染色结果（×100）

Figure 2. Staining results of HE and oil red O in liver tissues of mice（×100）

下载: 全尺寸图片幻灯片

注： a，OTU数；b，各水平微生物分类单元数统计图。

图 3 小鼠肠道菌群生物信息学分析

Figure 3. Bioinformatics analysis of the intestinal flora in mice

下载: 全尺寸图片幻灯片

注： a，稀疏曲线；b，Alpha多样性指数（与其他组比较，*P<0.05，**P<0.01，***P<0.001）；c，PCoA分析二维排序图；d，NMDS二维排序图。

图 4 小鼠肠道菌群多样性分析

Figure 4. Analysis of the diversity of intestinal flora in mice

下载: 全尺寸图片幻灯片

注： a，排名前20菌群门类；b，排名前20菌群纲类；c，排名前20菌群科类；d，排名前20菌群种类；e，各组物种组成热图；f，差异菌群。

图 5 小鼠肠道菌群的组成成分及差异分析

Figure 5. Analysis of the components and differences of the intestinal microbiota in mice

下载: 全尺寸图片幻灯片

注： a，每个比较组的基因表达量差异；b，热图；c，DAPA vs MAFLD；d，THP vs MAFLD。

图 6 小鼠mRNA表达量差异分析

Figure 6. Analysis of differences in mRNA expression levels in mice

下载: 全尺寸图片幻灯片

注： a，MAFLD组 vs Control组；b，DAPA组 vs MAFLD组；★P<0.05；★★P<0.01；★★★P<0.001。

图 7 小鼠肠道差异菌群与肝脏转录因子表达关联性分析

Figure 7. Correlation analysis of intestinal differential microbiota and liver transcription factor expression in mice

下载: 全尺寸图片幻灯片

表 1 小鼠生化指标情况

Table 1. Biochemical indicators of mice

指标	Control组	MAFLD组	DAPA组	THP组	DAPA+THP组	H值	P值
ALT（U/L）	85.34 （84.43～85.48）	87.33 （86.66～87.81）^1）	85.04 （84.81～85.40）^2）	85.01 （84.02～85.72）^2）	82.56 （81.60～83.59）^1）2）	24.15	<0.001
AST（U/L）	16.02 （15.90～16.15）	17.21 （16.97～17.35）^1）	16.21 （15.64～16.43）^2）	16.09 （15.84～16.43）^1）2）	15.93 （15.66～16.23）^2）	20.81	0.001
GSH（μmol/L）	65.63 （48.96～112.15）	10.42 （8.33～50.69）	54.34 （53.91～75.17）	85.42 （59.20～107.12）	53.13 （50.95～55.99）	9.37	0.053
MDA（nmol/mL）	29.39 （18.91～44.65）	261.28 （244.65～325.51）^1）	41.12 （30.52～53.25）^2）	318.91 （299.37～353.25）^1）	26.99 （15.49～30.64）	23.53	<0.001
SOD（U/mL）	19.88 （19.13～21.85）	18.62 （14.31～21.42）	22.06 （20.65～23.82）^2）	19.48 （18.91～20.38）	25.14 （23.73～26.42）^1）2）	19.35	0.001
TC（mmol/L）	6.89 （6.63～7.19）	8.81 （8.28～10.06）^1）	7.27 （6.59～7.63）^2）	9.82 （9.50～10.19）^2）	7.05 （6.70～8.46）^2）	20.37	0.001
TG（mmol/L）	0.96 （0.89～0.97）	1.56 （1.49～1.63）^1）	1.07 （0.90～1.14）^2）	1.04 （0.75～1.18）^1）	0.77 （0.73～0.78）^2）	18.16	0.001
HDL-C（mmol/L）	3.37 （3.14～3.44）	2.38 （2.21～2.59）^1）	3.00 （2.81～3.03）	2.84 （2.53～3.49）	2.52 （2.45～3.09）	14.43	0.006
LDL-C（mmol/L）	4.62 （4.47～4.99）	7.59 （7.11～8.27）^1）	4.89 （4.64～5.14）	4.62 （4.53～4.72）	6.19 （6.00～6.45）	22.79	0.000 1
注：与Control组比较，1）P<0.05；与MAFLD组比较，2）P<0.05。

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