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
Protection mechanism of Puerarin on H2O2-induced oxidative stress and mitochondrial damage in human podocytes under high glucose environment
- 北京中医药 2022年41卷第2期 页码：125-131
- 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：
朱青青，女，28岁，硕士，住院医师。研究方向：中医药防治糖尿病肾病。
钟逸斐，E-mail：yifeilily@126.com
- 基金信息：国家自然科学基金资助项目(81973772);上海市进一步加快中医药事业发展三年行动计划［ZY（2018-2020）-CCCX-2002-02］;上海市中医药领军人才计划［ZY（2018-2020）-RCPY-1007］;上海市临床重点专科—中医肾病科(shslczdzk04201);上海中医药大学附属龙华医院科研基金面上项目(2018YM01)
DOI：10.16025/j.1674-1307.2022.02.004
中图分类号：
扫 描 看 全 文
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
references
MAGLIANO DJ,ISLAM RM,BARR ELM,et al.Trends in incidence of total or type 2 diabetes:systematic review[J].BMJ Clin Res Ed,2019,366:l5003.
MOLITCH ME,ADLER AI,FLYVBJERG A,et al.Diabetic kidney disease:a clinical update from Kidney Disease:Improving Global Outcomes[J].Kidney Int,2015,87(1):20-30.
ZHANG LX,LONG JY,JIANG WS,et al.Trends in chronic kidney disease in China[J].N Engl J Med,2016,375(9):905-906.
THOMAS MC,COOPER ME,ZIMMET P.Changing epidemiology of type 2 diabetes mellitus and associated chronic kidney disease[J].Nat Rev Nephrol,2016,12(2):73-81.
TANAMAS SK,SAULNIER PJ,FUFAA GD,et al.Long-term effect of losartan on kidney disease in American indians with type 2 diabetes:a follow-up analysis of a randomized clinical trial[J].Diabetes Care,2016,39(11):2004-2010.
范婷,杨晓晖,李怡,等.糖尿病肾病的中医临床研究进展[J].北京中医药,2019,38(10):1036-1041.
景婧,庞博,王颖辉,等.糖尿病肾病Ⅲ-Ⅴ期中医证候演变规律研究[J].北京中医药,2011,30(12):888-890.
刘瑶,李伟.葛根素治疗糖尿病肾病的研究进展[J].中草药,2018,49(4):981-986.
DUAN HJ,LIU SX,ZHANG YJ,et al.Effects of puerarin on renal function,expressions of MMP-2 and TIMP-2 in diabetic rats[J].Acta Pharm Sin,2004,39(7):481-485.
WANG B,CHEN SB,YAN XF,et al.The therapeutic effect and possible harm of puerarin for treatment of stage III diabetic nephropathy:a meta-analysis[J].Altern Ther Health Med,2015,21(1):36-44.
钟羚君,谢治深,徐晓军.天然小分子化合物改善糖尿病肾病的研究进展[J].药学进展,2017,41(10):742-748.
SONG XB,LIU G,WANG ZY,et al.Puerarin protects against cadmium-induced proximal tubular cell apoptosis by restoring mitochondrial function[J].Chem Biol Interact,2016,260:219-231.
WANG ZK,ZHOU XL,SONG XB,et al.Alleviation of lead-induced apoptosis by puerarin via inhibiting mitochondrial permeability transition pore opening in primary cultures of rat proximal tubular cells[J].Biol Trace Elem Res,2016,174(1):166-176.
ZHONG YF,ZHANG XW,CAI XF,et al.Puerarin attenuated early diabetic kidney injury through down-regulation of matrix metalloproteinase 9 in streptozotocin-induced diabetic rats[J].PLoS One,2014,9(1):e85690.
LI X,CAI W,LEE K,et al.Puerarin attenuates diabetic kidney injury through the suppression of NOX4 expression in podocytes[J].Sci Rep,2017,7(1):14603.
LI XL,ZHU QQ,ZHENG R,et al.Puerarin attenuates diabetic nephropathy by promoting autophagy in podocytes[J].Front Physiol,2020,11:73.
MANIURA-WEBER K,GOFFART S,GARSTKA HL,et al.Transient overexpression of mitochondrial transcription factor A (TFAM) is sufficient to stimulate mitochondrial DNA transcription,but not sufficient to increase mtDNA copy number in cultured cells[J].Nucleic Acids Res,2004,32(20):6015-6027.
PATTI ME,BUTTE AJ,CRUNKHORN S,et al.Coordinated reduction of genes of oxidative metabolism in humans with insulin resistance and diabetes:Potential role of PGC1 and NRF1[J].Proc Natl Acad Sci USA,2003,100(14):8466-8471.
CAMPBELL CT,KOLESAR JE,KAUFMAN BA.Mitochondrial transcription factor A regulates mitochondrial transcription initiation,DNA packaging,and genome copy number[J].Biochim Biophys Acta,2012,1819(9/10):921-929.
HARBAUER AB,ZAHEDI RP,SICKMANN A,et al.The protein import machinery of mitochondria-a regulatory hub in metabolism,stress,and disease[J].Cell Metab,2014,19(3):357-372.
CHACINSKA A,KOEHLER CM,MILENKOVIC D,et al.Importing mitochondrial proteins:machineries and mechanisms[J].Cell,2009,138(4):628-644.
NEUPERT W,HERRMANN JM.Translocation of proteins into mitochondria[J].Annu Rev Biochem,2007,76:723-749.
OZBEK E.Induction of oxidative stress in kidney[J].Int J Nephrol,2012,2012:465897.
0
浏览量
0
下载量
0
CSCD
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备18046208号-2
京公网安备 11010102006710号 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies