保肾通络方含药血清调节线粒体自噬对高糖诱导下足细胞氧化损伤的影响

doi:10.3969/j.issn.1006-7795.2022.05.004

首都医科大学学报 ›› 2022, Vol. 43 ›› Issue (5): 687-693.doi: 10.3969/j.issn.1006-7795.2022.05.004

• 肾脏病基础与临床研究 • 上一篇下一篇

保肾通络方含药血清调节线粒体自噬对高糖诱导下足细胞氧化损伤的影响

王梦迪¹, 郭弋凡^1,2, 庞彦余¹, 刘羽飞^1,2, 赵文景^1*

1.首都医科大学附属北京中医医院肾病科,北京 100010;
2.北京中医药大学,北京 100029

收稿日期:2022-05-09 出版日期:2022-10-21 发布日期:2022-10-25
基金资助:
国家自然科学基金(81904105),北京市自然科学基金(7222271),首都医科大学附属北京中医医院职工助推项目(ZT202116)。

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).

摘要/Abstract

摘要： 目的观察保肾通络方含药血清对高糖培养下足细胞氧化损伤的影响,并对保肾通络方的作用机制进行探讨。方法以体外培养的肾小球足细胞为研究对象,给予高浓度葡萄糖刺激,采用CCK-8法分析足细胞活力以确定最佳模型及给药浓度,分为正常对照组、高糖组、保肾通络方组,采用鬼笔环肽进行细胞骨架染色,Western blotting法检测足细胞裂孔膜蛋白nephrin,自噬相关蛋白5(autophagy-related protein 5,ATG5)、ATG7、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)的表达,流式细胞术检测足细胞活性氧(reactive oxygen species,ROS)及线粒体膜电位,免疫荧光共标LC3B及线粒体,观察各组足细胞内线粒体自噬泡的生成情况。结果 30 mmol/L高糖干预48 h可显著降低足细胞活力(P＜0.05),保肾通络方含药血清可使高糖诱导的足细胞活力明显提升(P＜0.01);与正常对照组相比,高糖组足细胞骨架褶皱呈多边形,应力纤维束被破坏,ROS生成显著增多(P＜0.01),足细胞nephrin、ATG5、ATG7表达显著降低,LC3Ⅱ/LC3Ⅰ比值降低(P＜0.05),LC3B与线粒体共标减少,线粒体膜电位下降(P＜0.01);与高糖组相比,保肾通络方组足细胞骨架改善,ROS生成减少(P＜0.01),nephrin、ATG5、ATG7表达显著增加(P＜0.05),LC3Ⅱ/LC3Ⅰ比值上升(P＜0.05),LC3B与线粒体荧光共标增多,线粒体膜电位升高(P＜0.01)。结论保肾通络方含药血清能够减轻高糖诱导的足细胞氧化损伤,其作用可能与改善足细胞线粒体自噬有关。

关键词: 保肾通络方, 糖尿病肾脏疾病, 足细胞, 氧化损伤, 线粒体自噬

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

中图分类号:

王梦迪, 郭弋凡, 庞彦余, 刘羽飞, 赵文景. 保肾通络方含药血清调节线粒体自噬对高糖诱导下足细胞氧化损伤的影响[J]. 首都医科大学学报, 2022, 43(5): 687-693.

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.

参考文献

[1] Ruiz-Ortega M, Rodrigues-Diez R R, Lavoz C, et al. Special issue “diabetic nephropathy: diagnosis, prevention and treatment”[J]. J Clin Med, 2020, 9(3): 813.
[2] 中华医学会糖尿病学分会微血管并发症学组. 糖尿病肾病防治专家共识(2014年版)[J]. 中华糖尿病杂志, 2014, 6(11): 792-801.
[3] Podgórski P, Konieczny A, Lis Ł, et al. Glomerular podocytes in diabetic renal disease[J]. Adv Clin Exp Med, 2019, 28(12): 1711-1715.
[4] Sharma K. Obesity and diabetic kidney disease: role of oxidant stress and redox balance[J]. Antioxid Redox Signal, 2016, 25(4): 208-216.
[5] Fakhruddin S, Alanazi W, Jackson K E. Diabetes-induced reactive oxygen species: mechanism of their generation and role in renal injury[J]. J Diabetes Res, 2017, 2017: 8379327.
[6] Akbar S, Bellary S, Griffiths H R. Dietary antioxidant interventions in type 2 diabetes patients: a meta-analysis[J]. Bibliographic details, 2011, 11(2): 62-68.
[7] 孙雪艳, 赵文景, 刘志强, 等. 保肾通络方对糖尿病肾病Ⅲ～Ⅳ期患者临床疗效及足细胞损伤的影响[J]. 环球中医药, 2020, 13(1): 153-156.
[8] 沈存, 王悦芬, 孙雪艳, 等. 保肾通络方治疗2型糖尿病肾脏病气阴两虚兼血瘀证临床观察和足细胞保护机制研究[J]. 四川中医, 2020, 38(7): 142-146.
[9] Cui F Q, Wang Y F, Gao Y B, et al. Effects of BSF on podocyte apoptosis via regulating the ROS-mediated PI3K/AKT pathway in DN[J]. J Diabetes Res, 2019, 2019: 9512406.
[10] Audzeyenka I, Bieryńska A, Lay A C. Podocyte bioenergetics in the development of diabetic nephropathy: the role of mitochondria[J]. Endocrinology, 2022, 163(1): bqab234.
[11] Audzeyenka I, Rachubik P, Typiak M, et al. Hyperglycemia alters mitochondrial respiration efficiency and mitophagy in human podocytes[J]. Exp Cell Res, 2021, 407(1): 112758.
[12] Piwkowska A, Rogacka D, Audzeyenka I, et al. High glucose concentration affects the oxidant-antioxidant balance in cultured mouse podocytes[J]. J Cell Biochem, 2011, 112(6): 1661-1672.
[13] Lindblom R, Higgins G, Coughlan M, et al. Targeting mitochondria and reactive oxygen species-driven pathogenesis in diabetic nephropathy[J]. Rev Diabet Stud, 2015, 12(1/2): 134-156.
[14] Brady N R, Hamacher-Brady A, Westerhoff H V, et al. A wave of reactive oxygen species (ROS)-induced ROS release in a sea of excitable mitochondria[J]. Antioxid Redox Signal, 2006, 8(9/10): 1651-1665.
[15] Zhang P N, Zhou M Q, Guo J, et al. Mitochondrial dysfunction and diabetic nephropathy: nontraditional therapeutic opportunities[J]. J Diabetes Res, 2021, 2021: 1010268.
[16] Yang M, Li C R, Yang S K, et al. Mitophagy: a novel therapeutic target for treating DN[J]. Curr Med Chem, 2021, 28(14): 2717-2728.
[17] Sun A Q, Wei J, Childress C, et al. The E3 ubiquitin ligase NEDD4 is an LC3-interactive protein and regulates autophagy[J]. Autophagy, 2017, 13(3): 522-537.
[18] Periyasamy K M, Ranganathan U D, Tripathy S P, et al. Vitamin D - A host directed autophagy mediated therapy for tuberculosis[J]. Mol Immunol, 2020, 127: 238-244.
[19] Li W, Du M M, Wang Q Z, et al. FoxO1 promotes mitophagy in the podocytes of diabetic male mice via the PINK1/parkin pathway[J]. Endocrinology, 2017, 158(7): 2155-2167.

保肾通络方含药血清调节线粒体自噬对高糖诱导下足细胞氧化损伤的影响

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 7

编辑推荐

Metrics

本文评价

[1]	崔兴, 钟萍. 我的梦想是做“百年老店”——记首都医科大学三博脑科医院神经外科于春江教授[J]. 首都医科大学学报, 2021, 42(1): 167-169.
[2]	王明洋, 孙争宇, 张丽, 李雅莉, 李林, 张兰. 山茱萸环烯醚萜苷对脑缺血再灌注大鼠线粒体损伤的影响[J]. 首都医科大学学报, 2020, 41(3): 385-390.
[3]	郑晓敏, 杨芳远, 刘翠平, 杨金奎. 储存条件对尿触珠蛋白检测的影响[J]. 首都医科大学学报, 2020, 41(1): 21-26.
[4]	郑晓敏, 杨芳远, 刘翠平, 杨金奎. 触珠蛋白对早期糖尿病肾病患者肾功能损害的预测作用[J]. 首都医科大学学报, 2019, 40(1): 53-58.
[5]	王馨瑶, 耿建国, 高彦彬, 李娇阳, 周盛楠, 邹大威, 刘迎新. 芪卫颗粒对糖尿病肾病大鼠足细胞CD2AP表达的影响[J]. 首都医科大学学报, 2015, 36(2): 239-243.
[6]	陶真, 王荣亮, 赵海苹, 罗玉敏. miRNA-99a对过氧化氢诱导neuro-2a细胞氧化损伤的影响[J]. 首都医科大学学报, 2014, 35(3): 305-309.
[7]	李燕;张春来;卢峰;王立忠;姜玉凤. 高同型半胱氨酸血症对平滑肌细胞的氧化损伤[J]. 首都医科大学学报, 2012, 33(2): 251-254.