Screening and validation of key molecular targets for dihydromyricetin in ameliorating diabetic nephropathy

LIU Xingran; NIU Mengzhu; GAO Yuan; CHEN Dandan; ZHANG Baowen; LIU Dekun; LIU Xiaoqian; KOU Xianjuan

doi:10.16016/j.2097-0927.202507095

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Screening and validation of key molecular targets for dihydromyricetin in ameliorating diabetic nephropathy

Basic Medicine | 更新时间：2025-11-13

- Screening and validation of key molecular targets for dihydromyricetin in ameliorating diabetic nephropathy
- Journal of Army Medical University Vol. 47, Issue 21, Pages: 2663-2677(2025)
- 作者机构：
  
  1.武汉体育学院运动医学院，湖北武汉
  2.广西医科大学体育与健康学院，广西南宁
  3.哈尔滨体育学院运动科学与健康学院，黑龙江哈尔滨
  4.广西科技大学体育学院，广西柳州
  5.武汉体育学院运动训练监控湖北省重点实验室，湖北武汉
- 作者简介：
  
  KOU Xianjuan， E-mail： kouxianjuan@126.com
- 基金信息：
  
  the Key Project of Natural Science Foundation of Hubei Province(2024AFD242);the Guangxi Natural Science Foundation for Young Scholars(2025GXNSFBA069048);the Excellent Young and Middle-Aged Science and Technology Innovation Team Program for Higher Education Institutions of Hubei Province(T2024019);the Project of Basic Scientific Research Capability Improvement of Young and Middle-Aged Teachers in Guangxi Colleges and Universities(2023KY0365;2025KY1486);the Project of Integrated Sports and Medicine Health Services Base Guangxi Zhuang Autonomous Region(2022002)
- DOI：10.16016/j.2097-0927.202507095
  CLC： R285.5;R587.2;R692
- Received：28 July 2025，
  
  Revised：2025-10-22，
  
  Accepted：22 October 2025，
  
  Published：15 November 2025
- 稿件说明：
移动端阅览
刘醒然，牛梦竹，高原，等.二氢杨梅素改善糖尿病肾病的关键靶分子的筛选与验证［J］.陆军军医大学学报，2025，47（21）：2663-2677.DOI：10.16016/j.2097-0927.202507095.

LIU X R，NIU M Z，GAO Y，et al.Screening and validation of key molecular targets for dihydromyricetin in ameliorating diabetic nephropathy［J］.J Army Med Univ，2025，47（21）：2663-2677.DOI：10.16016/j.2097-0927.202507095.
刘醒然，牛梦竹，高原，等.二氢杨梅素改善糖尿病肾病的关键靶分子的筛选与验证［J］.陆军军医大学学报，2025，47（21）：2663-2677.DOI：10.16016/j.2097-0927.202507095. DOI：

LIU X R，NIU M Z，GAO Y，et al.Screening and validation of key molecular targets for dihydromyricetin in ameliorating diabetic nephropathy［J］.J Army Med Univ，2025，47（21）：2663-2677.DOI：10.16016/j.2097-0927.202507095. DOI：

摘要

目的

探讨二氢杨梅素（dihydromyricetin，DHM）改善糖尿病肾病（diabetic nephropathy，DN）的潜在作用机制，筛选其可能调控的关键分子靶点并进行验证。

方法

构建db/db小鼠DN模型，给予DHM（每天100 mg/kg，每周5 d）灌胃干预10周，采用病理学染色评估DHM对肾脏病理变化和纤维化的改善作用；从基因表达综合数据库（Gene Expression Omnibus，GEO）数据库中分别获取GSE161885与GSE270526数据集，结合GeneCards数据库筛选DN相关差异基因；采用PPI网络构建和分子对接预测DHM潜在靶点；利用Western blot和免疫荧光等技术检测DHM对db/db小鼠肾组织及高糖（high glucose，HG）诱导的人肾小管上皮细胞（HK-2）中细胞焦亡相关通路的影响，同时联合NLR家族PYD结构域含蛋白3（NLR family pyrin domain containing protein 3，NLRP3）特异性抑制剂MCC950以验证预测机制。

结果

体内实验显示，DHM显著改善db/db小鼠肾病理损伤，减轻肾小球肥大和系膜增生，显著减少Paller评分（

0.001）。免疫荧光显示肾脏α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）、Fibronectin、Ⅰ型胶原蛋白（Collagen Ⅰ）细胞荧光染色阳性信号明显减弱，肾脏组织Collagen Ⅲ、α-SMA和转化生长因子β1（transforming growth factor beta 1，TGF-β1）表达水平明显下降（

0.05）。基因筛选共获得16个DN相关差异基因。富集分析显示，这些基因主要富集于病毒蛋白、细胞因子-细胞因子受体相互作用、糖尿病并发症中的AGE-RAGE信号通路等相关通路，并在基因功能上主要涉及淋巴细胞介导免疫的正向调节、适应性免疫反应的正向调节和趋化因子活性。分子对接分析确定NLRP3为DHM潜在作用靶点。体内实验证实DHM明显下调膜打孔蛋白（gasdermin-D，GSDMD）荧光染色阳性信号，抑制NLRP3、半胱

氨酸蛋白酶1（Caspase 1）、活化的半胱氨酸蛋白酶1（Cleaved-Caspase 1）、白细胞介素18（interleukin 18，IL-18）和GSDMD等焦亡相关蛋白表达（

0.05）。体外研究进一步证实DHM可减轻高糖诱导的HK-2细胞纤维化和焦亡。体外研究进一步证实DHM与NLRP3特异性抑制剂MCC950均可减轻高糖诱导的HK-2细胞纤维化和焦亡。

结论

DHM可改善DN进展，其机制与抑制NLRP3炎症小体介导的细胞焦亡，进而减轻肾脏炎症和纤维化有关。

Abstract

Objective

To investigate the potential mechanism by which dihydromyricetin (DHM) ameliorates diabetic nephropathy (DN)

and to screen and validate its possible key molecular targets.

Methods

A DN model was established using db/db mice

and 100 mg/(kg·d) DHM was administered

via

gavage 5 d per week for totally 10 weeks. Renal morphological changes were observed after staining to evaluate the effects of DHM. GSE161885 and GSE270526 datasets were obtained from the Gene Expression Omnibus (GEO) database and analyzed in combination with the GeneCards database to screen for DN-related differentially expressed genes (DEGs). Protein-protein interaction (PPI) network and molecular docking were employed to predict potential DHM targets. Western blotting and immunofluorescence staining were performed to detect the effects of DHM on pyroptosis-related pathways in the renal tissues of db/db mice and in high glucose (HG)-induced human renal tubular epithelial cells (HK-2). The specific NLR family pyrin domain containing protein 3 (NLRP3) inhibitor MCC950 was also used to validate the predicted mechanism.

Results

In vivo

experiments showed that DHM significantly ameliorated renal pathological damage in db/db mice

alleviated glomerular hypertrophy and mesangial expansion

and markedly reduced Paller scores (

0.001). Immunofluorescence staining revealed significantly weakened fluorescence signals for α-smooth muscle actin (α-SMA)

fibronectin

and collagen Ⅰ in renal tissues. Western blot results showed that the expression levels

of collagen Ⅰ

collagen Ⅲ

α-SMA

and transforming growth factor beta 1 (TGF-β1) were significantly decreased (

0.05). A total of 16 DN-related DEGs were identified. Enrichment analysis revealed that these genes were primarily enriched in pathways such as viral protein interactions

cytokine-cytokine receptor interaction

and the AGE-RAGE signaling pathway in diabetic complications

and were primarily involved in gene functions such as the positive regulation of lymphocyte-mediated immunity

positive regulation of adaptive immune response

and chemokine activity. Molecular docking confirmed NLRP3 as a potential target of DHM.

In vivo

validation showed that DHM significantly down-regulated gasdermin-D (GSDMD) fluorescence signals and inhibited the expression of pyroptosis-related proteins including NLRP3

Caspase 1

Cleaved-Caspase 1

interleukin 18 (IL-18)

and GSDMD (

0.05).

In vitro

studies further confirmed that both DHM and the specific NLRP3 inhibitor MCC950 alleviate high glucose-induced fibrosis and pyroptosis in HIC-2 cells.

Conclusion

DHM can ameliorate the progression of DN

and its mechanism is related to inhibiting NLRP3 inflammasome-mediated pyroptosis

thereby alleviating renal inflammation and fibrosis.

关键词

Keywords

references

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