Effect of Huangkui extract powder on the expression of nephrin and podocin in podocyte induced by high glucose

中国基层医药 Pub Date : 2020-02-15 DOI:10.3760/CMA.J.ISSN.1008-6706.2020.04.012

Chunli Yu, Sisi Dai, Junli Gao

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Abstract

Objective To investigate the effect of Huangkui extract powder (HK) on the expression of nephrin and podocin proteins in mouse podocytes induced by high glucose, which is involved in the treatment of diabetic nephropathy (DN). Methods Cultured mouse podocytes (MPC5) were incubated in high glucose and HK at 5.6 mmol/L NG, 5.6 mmol/L NG+ 0.45 g/L HK, 25 mmol/L HG, 25 mmol/L HG+ 0.45 g/L HK, respectively.The 5.6 mmol/L NG group was used as normal control.After 24 hours of intervention, we detected podocyte apoptosis by Annexin-V FITC/PI double staining, measured the mRNA and protein expression of nephrin and podocin by qRT-PCR and Western blot. Results Compared with the control group (5.6 mmol/L, NG), the apoptosis rate of podocytes in the high glucose concentration group (25 mmol/L, HG) was significantly higher[(20.39±0.03)% vs.(17.70±0.91)%, t=2.947, P<0.05)]. The apoptosis rate of podocytes in the 25 mmol/L HG+ 0.45 g/L HK group was significantly lower than that in the 25 mmol/L HG group[(11.96±1.11)% vs.(20.39±0.03)%, t=7.586, P<0.01]. The results of qRT-PCR and Western blot showed that the expression of nephrin and podocin was significantly inhibited by high glucose concentration compared with the control group[(0.489±0.040) vs.(0.721±0.022), t=4.992, P<0.01; (0.387±0.014) vs.(0.778±0.036), t=10.050, P<0.01], and the expression of podocin and nephrin was increased by appropriate concentration of HK [(0.603±0.013) vs.(0.489±0.040), t=2.653, P<0.05; (0.640±0.024) vs.(0.387±0.014), t=8.946, P<0.01]. Conclusion Podocyte apoptosis can be induced by prolonged high glucose treatment, but a certain concentration of HK can inhibit podocyte death induced by high glucose.The possible mechanism is that HK may inhibit the apoptosis of podocytes by regulating the expression of podocin and nephrin in podocytes at high glucose concentration, thus plays a protective role on podocytes. Key words: Diabetic nephropathy; Huangkui extract powder; Glucose; Podocyte; Nephrin; Podocin; Gene regulation; Apoptosis

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黄葵提取物粉对高糖诱导足细胞中肾素和足素表达的影响

目的探讨黄葵提取物散(HK)对高糖诱导小鼠足细胞nephrin和podocin蛋白表达的影响，探讨其对糖尿病肾病(DN)的治疗作用。方法将培养的小鼠足细胞(MPC5)分别培养于5.6 mmol/L NG、5.6 mmol/L NG+ 0.45 g/L HK、25 mmol/L HG、25 mmol/L HG+ 0.45 g/L HK的高糖和HK环境中。以5.6 mmol/L NG组为正常对照组。干预24h后，采用Annexin-V FITC/PI双染色检测足细胞凋亡，采用qRT-PCR和Western blot检测肾素和足素mRNA和蛋白的表达。结果高糖浓度组(25 mmol/L, HG)足细胞凋亡率显著高于对照组(5.6 mmol/L, NG)[(20.39±0.03)% vs(17.70±0.91)%，t=2.947, P<0.05]。25 mmol/L HG+ 0.45 g/L HK组足细胞凋亡率显著低于25 mmol/L HG组[(11.96±1.11)% vs(20.39±0.03)%，t=7.586, P<0.01]。qRT-PCR和Western blot结果显示，与对照组相比，高葡萄糖浓度显著抑制nephrin和podocin的表达[(0.489±0.040)vs(0.721±0.022)]，t=4.992, P<0.01;(0.387±0.014)比(0.778±0.036)，t=10.050, P<0.01]，适当浓度HK可提高podocin、nephrin的表达[(0.603±0.013)比(0.489±0.040)，t=2.653, P<0.05];(0.640±0.024)和(0.387±0.014),t = 8.946, P < 0.01)。结论长时间高糖处理可诱导足细胞凋亡，但一定浓度的HK可抑制高糖诱导的足细胞死亡。其机制可能是HK通过调节高葡萄糖浓度下足细胞中podocin和nephrin的表达抑制足细胞凋亡，从而对足细胞起到保护作用。关键词:糖尿病肾病;黄葵浸膏粉;葡萄糖;足突细胞;Nephrin;Podocin;基因调控;细胞凋亡

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来源期刊

中国基层医药

CiteScore

0.10

自引率

0.00%

发文量

32251

期刊介绍： Since its inception, the journal "Chinese Primary Medicine" has adhered to the development strategy of "based in China, serving the grassroots, and facing the world" as its publishing concept, reporting a large amount of the latest medical information at home and abroad, prospering the academic field of primary medicine, and is praised by readers as a medical encyclopedia that updates knowledge. It is a core journal in China's medical and health field, and its influence index (CI) ranks Q2 in China's academic journals in 2022. It was included in the American Chemical Abstracts in 2008, the World Health Organization Western Pacific Regional Medical Index (WPRIM) in 2009, and the Japan Science and Technology Agency Database (JST) and Scopus Database in 2018, and was included in the Wanfang Data-China Digital Journal Group and the China Academic Journal Comprehensive Evaluation Database.