[Effects of ferulic acid on inflammation and autophagy levels in glomerular mesangial cells induced by high glucose].

Qing Fang, Ru-Yu Ma, Ying-Hao He, Min-You Qi
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Abstract

Objective: To investigate the protective effects and possible mechanisms of ferulic acid on diabetic nephropathy by observing the effects of ferulic acid on the level of inflammation and autophagy in glomerular mesangial cells induced by high glucose.

Methods: SV40 MES 13 cells were cultured and randomly divided into the following groups: normal group (Control, 5.6 mmol/L glucose), mannitol group (Man, 30 mmol/L mannitol), high glucose group (HG, 30 mmol/L glucose), ferulic acid group (FA, 30 mmol/L glucose + 12.5, 25, 50, 100, 200 μmol/L ferulic acid), and the proliferation of SV40 MES 13 cells in each group was observed by MTT method. The levels of tumour necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1) and interleukin 1β(IL-1β)in cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of NLRP3, IL-1β, LC3-II/I and p62 proteins in SV40 MES 13 cells were detected by Western blot.

Results: ①The proliferative activity of SV40 MES 13 cells was significantly higher in the HG group compared to the control group (P<0.01), while the proliferative activity of SV40 MES 13 cells was decreased to different degrees in the FA group compared to the HG group (P<0.05~0.01). ②Compared to the control group, the levels of TNF-α, MCP-1 and IL-1β were increased significantly in the cell supernatant of HG group (P<0.01). Compared with the HG group, the levels of TNF-α, MCP-1 and IL-1β were decreased significantly in the FA group (P<0.01). ③Compared with the control group, LC3-II/Ⅰ protein expression was decreased in the HG group, while the levels of p62, NLRP3 and IL-1β protein were increased significantly (P<0.01). Compared with the HG group, the expression of LC3-II/Ⅰ protein was elevated significantly (P<0.05) in the FA group, while the levels of p62, NLRP3 and IL-1β protein in the FA group were decreased significantly (P< 0.01).

Conclusion: FA can inhibit the abnormal proliferation of SV40 MES 13 cells induced by high glucose. FA can protect glomerular mesangial cells by inhibiting inflammation and increasing the level of autophagy.

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[阿魏酸对高糖诱导肾小球系膜细胞炎症和自噬水平的影响]。
目的:通过观察阿魏酸对高糖诱导的肾小球系膜细胞炎症和自噬水平的影响,探讨阿魏酸对糖尿病肾病的保护作用及其可能机制。方法:培养SV40 MES 13细胞,随机分为正常组(Control, 5.6 mmol/L葡萄糖)、甘甘醇组(Man, 30 mmol/L甘甘醇)、高糖组(HG, 30 mmol/L葡萄糖)、阿魏酸组(FA, 30 mmol/L葡萄糖+ 12.5、25、50、100、200 μmol/L阿魏酸),MTT法观察各组SV40 MES 13细胞的增殖情况。采用酶联免疫吸附法(ELISA)检测细胞上清液中肿瘤坏死因子-α (TNF-α)、单核细胞趋化蛋白-1 (MCP-1)和白细胞介素1β(IL-1β)水平。Western blot检测sv40mes 13细胞中NLRP3、IL-1β、LC3-II/I和p62蛋白的表达。结果:①HG组SV40 MES 13细胞的增殖活性明显高于对照组(P<0.01), FA组SV40 MES 13细胞的增殖活性明显低于HG组(P<0.05~0.01)。②与对照组相比,HG组细胞上清液中TNF-α、MCP-1、IL-1β水平显著升高(P<0.01)。与HG组相比,FA组TNF-α、MCP-1、IL-1β水平显著降低(P<0.01)。③与对照组比较,HG组LC3-II/Ⅰ蛋白表达降低,p62、NLRP3、IL-1β蛋白水平显著升高(P<0.01)。与HG组比较,FA组LC3-II/Ⅰ蛋白表达水平显著升高(P<0.05),而FA组p62、NLRP3、IL-1β蛋白表达水平显著降低(P< 0.01)。结论:FA可抑制高糖诱导的sv40mes 13细胞异常增殖。FA可通过抑制炎症和增加自噬水平来保护肾小球系膜细胞。
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