Research of Baoshentongluo formula-medicated serum on regulating mitophagy and reducing oxidative damage of podocytes induced by high glucose

doi:10.3969/j.issn.1006-7795.2022.05.004

Journal of Capital Medical University ›› 2022, Vol. 43 ›› Issue (5): 687-693.doi: 10.3969/j.issn.1006-7795.2022.05.004

• Clinical and Basic Research in Nephrology • Previous Articles Next Articles

Research of Baoshentongluo formula-medicated serum on regulating mitophagy and reducing oxidative damage of podocytes induced by high glucose

Wang Mengdi¹, Guo Yifan^1,2, Pang Yanyu¹, Liu Yufei^1,2, Zhao Wenjing^1*

1. Department of Nephrology, Beijing Hospital of Traditional Chinese Medicine,Capital Medical University, Beijing 100010,China;
2. Beijing University of Chinese Medicine, Beijing 100029, China

Received:2022-05-09 Online:2022-10-21 Published:2022-10-25
Contact: * E-mail:wenjingz@263.net
Supported by:
National Natural Science Foundation of China (81904105),Natural Science Foundation of Beijing(7222271), Staff Boost Project of Beijing Hospital of Traditional Chinese Medicine, Capital Medical University(ZT202116).

Abstract

Abstract: Objective To observe the effect of Baoshentongluo formula-medicated serum on oxidative damage of podocytes cultured with high glucose, and explore its underlying mechanism. Methods The podocytes in vitro were used as the research object, and stimulated with high concentration of glucose. CCK-8 method was used to determine optimal model and drug concentration, and then the podocytes were divided into the normal control group, high glucose group and Baoshentongluo formula group. Cytoskeleton of podocytes was stained with phalloidin.The expression levels of podocyte-specific protein nephrin and autophagy-related proteins autophagy-related protein 5(ATG5), ATG7 and microtubule-associated protein 1 light chain 3(LC3) was detected with Western blotting.Flow cytometry was used to detect the level of reactive oxygen species (ROS) and mitochondrial membrane potential of podocytes. Colocalization of LC3B and mitochondria were examined with immunofluorescence to observe the generation of mitochondrial autophagy vesicles in podocytes of each group. Results The podocyte viability was significantly decreased with 30 mmol/L high glucose intervention for 48 h (P<0.05), and Baoshentongluo formula-medicated serum could significantly increase the viability of high glucose-induced podocyte (P<0.01).Compared with normal control group, the podocytes of high glucose group were demonstrated with folded skeleton into a polygon and destroyed stress fiber bundles with increased ROS production (P<0.01), reduced expression of nephrin, ATG5, ATG7 and LC3Ⅱ/LC3Ⅰ ratio (P<0.05), decreased co-labeling of LC3B with mitochondria, lowerer mitochondrial membrane potential (P<0.01). Compared with high glucose group, the podocytes of Baoshentongluo formula group showed improved podocyte structure,decreased production of ROS (P＜0.01), elevated expression of nephrin, ATG5, ATG7 and LC3Ⅱ/LC3Ⅰ ratio (P＜0.05) ,increased co-labeling of LC3B with mitochondria, higher mitochondrial membrane potential (P＜0.01). Conclusion Baoshentongluo formula-medicated serum can alleviate oxidative damage to podocytes induced by high glucose and the mechanism may be related to improvement of mitophagy.

Key words: Baoshentongluo formula, diabetic kidney disease, podocyte, oxidative damage, mitophagy

CLC Number:

Wang Mengdi, Guo Yifan, Pang Yanyu, Liu Yufei, Zhao Wenjing. Research of Baoshentongluo formula-medicated serum on regulating mitophagy and reducing oxidative damage of podocytes induced by high glucose[J]. Journal of Capital Medical University, 2022, 43(5): 687-693.

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Research of Baoshentongluo formula-medicated serum on regulating mitophagy and reducing oxidative damage of podocytes induced by high glucose

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