Protection mechanism of Puerarin on H<sub>2</sub>O<sub>2</sub>-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment

Qing-qing ZHU; Xue-ling LI; Jia-yi YAN; Yi-chen FAN; Ni LIU; Yi-fei ZHONG

doi:10.16025/j.1674-1307.2022.02.004

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Protection mechanism of Puerarin on H₂O₂-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment

更新时间：2023-02-15

- Protection mechanism of Puerarin on H₂O₂-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment
- Beijing Journal of Traditional Chinese Medicine Vol. 41, Issue 2, Pages: 125-131(2022)
- 作者机构：
  
  1.上海中医药大学附属龙华医院肾内科，上海 200032
- 作者简介：
- 基金信息：
- DOI：10.16025/j.1674-1307.2022.02.004
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朱青青，李雪玲，严佳怡，等.葛根素对高糖环境下H₂O₂诱导人足细胞氧化应激及线粒体损伤的保护机制［J］.北京中医药，2022，41（2）：125-131.

ZHU Qing-qing,LI Xue-ling,YAN Jia-yi,et al.Protection mechanism of Puerarin on H₂O₂-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment[J]. Beijing Journal of Traditional Chinese Medicine,2022,41(02):125-131.
朱青青，李雪玲，严佳怡，等.葛根素对高糖环境下H₂O₂诱导人足细胞氧化应激及线粒体损伤的保护机制［J］.北京中医药，2022，41（2）：125-131. DOI： 10.16025/j.1674-1307.2022.02.004.

ZHU Qing-qing,LI Xue-ling,YAN Jia-yi,et al.Protection mechanism of Puerarin on H₂O₂-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment[J]. Beijing Journal of Traditional Chinese Medicine,2022,41(02):125-131. DOI： 10.16025/j.1674-1307.2022.02.004.

摘要

目的,2,探讨葛根素对高糖环境下H,2,O,2,诱导的人永生型足细胞氧化应激及线粒体损伤的影响及其作用机制。,方法,2,将足细胞分组给予高糖（30 mmol/L Glucose）及不同浓度H,2,O,2,刺激，同时加不同浓度葛根素干预48 h。分组如下：H0组（H,2,O,2, 0 μmol/L）、H100组（H,2,O,2, 100 μmol/L）、H100+P3.3组（H,2,O,2, 100 μmol/L+葛根素3.3 μmol/L）、H100+P10组（H,2,O,2, 100 μmol/L+葛根素10 μmol/L）、H200组（H,2,O,2, 200 μmol/L）、H200+P3.3组（H,2,O,2, 200 μmol/L+葛根素3.3 μmol/L）、H200+P10组（H,2,O,2, 200 μmol/L+葛根素10 μmol/L）。CCK-8检测细胞存活率，Western blotting检测TOMM22的蛋白表达；提取基因组DNA，qPCR检测mtDNA的表达；提取RNA，qPCR检测TFAM、NRF1 mRNA表达；测定各组ATP水平、ROS水平，并评价各组足细胞线粒体呼吸功能。,结果,2,与H0组相比，H100、H200组ROS水平升高（,P,<,0.01），H100、H100+P3.3组mtDNA拷贝数均减少（,P,<,0.01），TFAM mRNA表达降低，基础呼吸、质子渗漏、最大耗氧量、非线粒体氧消耗均升高（,P,<,0.05）；与H100组相比，H100+P3.3组ROS减少（,P,<,0.05），TFAM mRNA、NRF1 mRNA表达均增加（,P,<,0.05，,P,<,0.01），ATP生成增加（,P,<,0.05），基础呼吸、质子渗漏均降低（,P,<,0.05），而H100+P10组mtDNA拷贝数增加（,P,<,0.05），NRF1 mRNA、基础呼吸、质子渗漏、非线粒体消耗均升高（,P,<,0.05）；与H100+P3.3组相比，H100+P10组mtDNA拷贝数明显增加（,P,<,0.01），基础呼吸、质子渗漏、非线粒体氧消耗、ATP水平均升高（,P,<,0.05）；与H200组相比，H200+P3.3、H200+P10组 ROS水平明显降低（,P,<,0.01），H200+P10组NRF1 mRNA表达增加（,P,<,0.01）、非线粒体氧消耗增加（,P,<,0.05）；与H200+P3.3组相比，H200+P10组ROS水平增多（,P,<,0.01），TFAM mRNA表达增加（,P,<,0.05）、NRF1 mRNA表达增加（,P,<,0.01）。各组TOMM22蛋白表达、备用呼吸能力、耦合效率和备用呼吸率比较，差异无统计学意义（,P,>,0.05）。,结论,2,葛根素能部分缓解高糖及H,2,O,2,诱导的足细胞氧化应激及线粒体损伤。

Abstract

Objective,2,To explore the effect of puerarin on oxidative stress and mitochondrial damage in human immortal podocytes induced by H,2,O,2, under high glucose environment and its mechanism.,Methods,2,Human immortal podocytes were stimulated with different concentrations of H,2,O,2, in a high-glucose environment, and intervened by different concentrations of puerarin for 48 hours. The groups were divided as follows: group H0 (H,2,O,2, 0 μmol/L), group H100 (H,2,O,2, 100 μmol/L), group H100+P3.3(H,2,O,2 ,100 μmol/L+ puerarin 3.3 μmol/L), group H100+ P10 (H,2,O,2, 100 μmol/L+ puerarin 10 μmol/L), group H200 (H,2,O,2, 200 μmol/L), group H200+P3.3(H,2,O,2, 200 μmol/L+ puerarin 3.3 μmol/L), group H200+P10(H,2,O,2, 200 μmol/L+puerarin 10 μmol/L). CCK-8 was used to detect cell viability. Western blotting was used to detect the protein expression of TOMM22; Genomic DNA was extracted and qPCR was used to detect the expression of mtDNA; RNA was extracted and qPCR was used to detect the mRNA expression of TFAM and NRF1;the levels of ATP and ROS in each group were measured and the mitochondrial respiratory function were evaluated.,Results,2,Compared with H0 group, ROS level was increased in H100 and H200 groups (,P,<,0.01), mtDNA copy number was decreased in H100 and H100+P3.3 groups (,P,<,0.01), TFAM mRNA expression was decreased, basal respiration, proton leakage, maximum oxygen consumption and non-mitochondrial oxygen consumption were increased (,P,<,0.05). Compared with H100 group, ROS in H100+P3.3 group was decreased (,P,<,0.05), TFAM mRNA and NRF1 mRNA expression were increased (,P,<,0.05,P,<,0.01), ATP production was increased (,P,<,0.05), basal respiration and proton leakage were decreased (,P,<,0.05) in group H100+P3.3. In H100+P10 group, mtDNA copy number was increased (,P,<,0.05), NRF1 mRNA, basal respiration, proton leakage and non-mitochondrial consumption were increased (,P,<,0.05). Compared with H100+P3.3 group, the mtDNA copy number of H100+P10 group was significantly increased (,P,<,0.01), basal respiration, proton leakage, non-mitochondrial oxygen consumption and ATP levels were increased (,P,<,0.05). Compared with H200 group, ROS levels in H200+P3.3 and H200+P10 groups were significantly decreased (,P,<,0.01), NRF1 mRNA expression was increased (,P,<,0.01) and non-mitochondrial oxygen consumption was increased (,P,<,0.05) in H200+P10 group. Compared with H200+P3.3 group, ROS level in H200+P10 group increased (,P,<,0.01), TFAM mRNA expression increased (,P,<,0.05), NRF1 mRNA expression increased (,P,<,0.01). There were no statistically significant differences in TOMM22 protein expression, reserve respiratory capacity, coupling efficiency and reserve respiratory rate among all groups (,P,>,0.05).,Conclusion,2,Puerarin can partially relieve podocyte oxidative stress and mitochondrial damage induced by high glucose and H,2,O,2,.

关键词

葛根素糖尿病性肾脏疾病氧化应激线粒体损伤

Keywords

Puerarindiabetic kidney diseaseoxidative stressmitochondrial damage

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School of Traditional Chinese Medicine， Capital Medical University

Department of Nephrology， Beijing Hospital of Traditional Chinese Medicine， Affiliated to the Capital Medical University

Beijing University of Chinese Medicine

Dongzhimen Hospital of Beijing University of Chinese Medicine

Institute of Traditional Chinese Medicine Encephalopathy，Beijing University of Chinese Medicine，Encephalopathy Department

⁰

Protection mechanism of Puerarin on H2O2-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment

DOI：10.16025/j.1674-1307.2022.02.004

摘要

Abstract

关键词

Keywords

references

Protection mechanism of Puerarin on H₂O₂-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment