Qufeng tongluo decoction decreased proteinuria in diabetic mice by protecting podocytes via promoting autophagy

IF 3.3 3区医学 Q1 INTEGRATIVE & COMPLEMENTARY MEDICINE Journal of Traditional and Complementary Medicine Pub Date : 2023-11-21 DOI:10.1016/j.jtcme.2023.11.007

Boran Ni , Yao Xiao , Ruojun Wei , Weijing Liu , Liwei Zhu , Yifan Liu , Zhichao Ruan , Jiamu Li , Shidong Wang , Jinxi Zhao , Weijun Huang

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Abstract

Background

Diabetic kidney disease (DKD) is one of diabetic complications, which has become the leading cause of end-stage kidney disease. In addition to angiotensin-converting enzyme inhibitor/angiotensin II receptor blocker(ACEI/ARB) and sodium-glucose cotransporter-2 inhibitor (SGLT2i), traditional Chinese medicine (TCM) is an effective alternative treatment for DKD. In this study, the effect of Qufeng Tongluo (QFTL) decoction in decreasing proteinuria has been observed and its mechanism has been explored based on autophagy regulation in podocyte.

Methods

In vivo study, db/db mice were used as diabetes model and db/m mice as blank control. Db/db mice were treated with QFTL decoction, rapamycin, QFTL + 3-Methyladenine (3-MA), trehalose, chloroquine (CQ) and QFTL + CQ. Mice urinary albumin/creatinine (UACR), nephrin and autophagy related proteins (LC3 and p62) in kidney tissue were detected after intervention of 9 weeks. Transcriptomics was operated with the kidney tissue from model group and QFTL group. In vitro study, mouse podocyte clone-5 (MPC-5) cells were stimulated with hyperglycemic media (30 mmol/L glucose) or cultured with normal media. High-glucose-stimulated MPC-5 cells were treated with QFTL freeze-drying powder, rapamycin, CQ, trehalose, QFTL+3-MA and QFTL + CQ. Cytoskeletal actin, nephrin, ATG-5, ATG-7, Beclin-1, cathepsin L and cathepsin B were assessed. mRFP-GFP-LC3 was established by stubRFP-sensGFP-LC3 lentivirus transfection.

Results

QFTL decoction decreased the UACR and increased the nephrin level in kidney tissue and high-glucose-stimulated podocytes. Autophagy inhibitors, including 3-MA and chloroquine blocked the effects of QFTL decoction. Further study showed that QFTL decoction increased the LC3 expression and relieved p62 accumulation in podocytes of db/db mice. In high-glucose-stimulated MPC-5 cells, QFTL decoction rescued the inhibited LC3 and promoted the expression of ATG-5, ATG-7, and Beclin-1, while had no effect on the activity of cathepsin L and cathepsin B. Results of transcriptomics also showed that 51 autophagy related genes were regulated by QFTL decoction, including the genes of ATG10, SCOC, ATG4C, AMPK catalytic subunit, PI3K catalytic subunit, ATG3 and DRAM2.

Conclusion

QFTL decoction decreased proteinuria and protected podocytes in db/db mice by regulating autophagy.

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曲风通络汤通过促进自噬保护荚膜细胞，从而减少糖尿病小鼠的蛋白尿

背景糖尿病肾病（DKD）是糖尿病并发症之一，已成为终末期肾病的主要病因。除了血管紧张素转换酶抑制剂/血管紧张素 II 受体阻滞剂（ACEI/ARB）和钠-葡萄糖共转运体-2 抑制剂（SGLT2i）外，中药也是治疗 DKD 的有效替代疗法。本研究观察了曲风通络汤（QFTL）降低蛋白尿的作用，并基于荚膜细胞的自噬调节探讨了其作用机制。Db/db 小鼠分别接受 QFTL 煎剂、雷帕霉素、QFTL + 3-甲基腺嘌呤（3-MA）、曲哈糖、氯喹（CQ）和 QFTL + CQ 治疗。干预 9 周后，检测了肾组织中的小鼠尿白蛋白/肌酐（UACR）、肾素和自噬相关蛋白（LC3 和 p62）。对模型组和 QFTL 组的肾组织进行了转录组学研究。在体外研究中，小鼠荚膜细胞克隆-5（MPC-5）细胞用高血糖培养基（30 mmol/L葡萄糖）刺激或用正常培养基培养。用 QFTL 冻干粉、雷帕霉素、CQ、妥哈糖、QFTL+3-MA 和 QFTL + CQ 处理高血糖刺激的 MPC-5 细胞。结果QFTL煎剂降低了肾组织和高葡萄糖刺激的荚膜细胞中的UACR，提高了肾素水平。3-MA和氯喹等自噬抑制剂阻断了QFTL煎剂的作用。进一步的研究表明，蟾酥煎剂可增加 LC3 的表达，并缓解 p62 在 db/db 小鼠荚膜细胞中的积累。在高葡萄糖刺激的 MPC-5 细胞中，芪苈强心煎剂能挽救被抑制的 LC3，促进 ATG-5、ATG-7 和 Beclin-1 的表达，但对 cathepsin L 和 cathepsin B 的活性没有影响。转录组学研究结果表明，51个自噬相关基因受QFTL煎剂调控，包括ATG10、SCOC、ATG4C、AMPK催化亚基、PI3K催化亚基、ATG3和DRAM2。

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来源期刊

Journal of Traditional and Complementary Medicine Medicine-Complementary and Alternative Medicine

CiteScore

9.30

自引率

6.70%

发文量

审稿时长

66 days

期刊介绍： eJTCM is committed to publish research providing the biological and clinical grounds for using Traditional and Complementary Medical treatments as well as studies that demonstrate the pathophysiological and molecular/biochemical bases supporting the effectiveness of such treatments. Review articles are by invitation only. eJTCM is receiving an increasing amount of submission, and we need to adopt more stringent criteria to select the articles that can be considered for peer review. Note that eJTCM is striving to increase the quality and medical relevance of the publications.