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目的:探讨二甲双胍对高糖状态下大鼠肾小球系膜细胞增殖的作用以及可能的作用机制。方法:体外培养大鼠肾小球系膜细胞(HBZY-1),①分为正常对照(NC)组(葡萄糖浓度5.6 mmol/L)、NC+二甲双胍组(葡萄糖浓度5.6 mmol/L+二甲双胍浓度200 mmol/L)、高糖(HG)组(葡萄糖浓度30 mmol/L)、HG+二甲双胍组(葡萄糖浓度30 mmol/L+二甲双胍,浓度分别是50、100、200、400 mmol/L),作用时间分别为6、12、24 h。MTT法检测各组HBZY-1细胞增殖的情况;②分为NC组、NC+二甲双胍组、HG组和HG+二甲双胍组(二甲双胍浓度200 mmol/L),作用时间12 h,用Western blot;法检测各组细胞内转化生长因子-β1(TGF-β1)、p-AMPK和AMPK蛋白表达的变化。结果:①与NC组相比,高糖刺激12 h后HBZY-1增殖明显,差异有显著性(P<0.01);与HG组相比,HG加二甲双胍干预12 h后,随着二甲双胍的浓度逐渐增加,对HBZY-1增殖的抑制程度逐渐加大,差异均有显著性(P均<0.01);NC+二甲双胍组与NC组相比,两组之间HBZY-1增殖差异没有显著性(P>0.05);②与NC组相比,高糖刺激12 h后HBZY-1细胞TGF-β1的表达增高而p-AMPK表达水平明显下降,差异均具有显著性(P均<0.01)。与HG组相比,加用二甲双胍干预后,TGF-β1的表达水平有明显下降,p-AMPK表达水平有显著升高,差异均具有显著性(P均<0.01)。4组之间AMPK表达差异无显著性(P>0.05)。结论:高糖环境下HBZY-1增殖明显,二甲双胍可能通过增加肾小球系膜细胞p-AMPK的表达、降低TGF-β1的表达抑制HBZY-1的增殖。
Objective: To investigate the effect of metformin on the proliferation of rat mesangial cells under high glucose and its possible mechanism. Methods: Rat mesangial cells (HBZY-1) were cultured in vitro. ① The rats were divided into normal control group (glucose concentration 5.6 mmol / L), NC + metformin group (glucose concentration 5.6 mmol / / L, high glucose (glucose 30 mmol / L), HG + metformin (glucose 30 mmol / L + metformin at concentrations of 50, 100, 200 and 400 mmol / L, respectively) 6,12,24 h. MTT assay was used to detect the proliferation of HBZY-1 cells in each group; ②The rats were divided into NC group, NC + metformin group, HG group and metformin group (metformin concentration 200 mmol / L) for 12 h, Changes of intracellular expression of transforming growth factor-β1 (TGF-β1), p-AMPK and AMPK protein. Results: ①Compared with NC group, the proliferation of HBZY-1 was significantly increased after 12 h of high glucose stimulation (P <0.01); Compared with HG group, after treated with HG plus metformin for 12 h, with the increase of metformin concentration (P <0.01). There was no significant difference in the proliferation of HBZY-1 between NC + metformin group and NC group (P> 0.05) > 0.05). ②Compared with NC group, the expression of TGF-β1 in HBZY-1 cells was increased and the expression of p-AMPK in HBZY-1 cells was significantly decreased after 12 h of high glucose stimulation compared with NC group (all P <0.01). Compared with HG group, the expression of TGF-β1 was significantly decreased and the expression of p-AMPK was significantly increased after treatment with metformin (all P <0.01). There was no significant difference in AMPK expression between the four groups (P> 0.05). CONCLUSION: HBzY-1 can proliferate in high glucose environment. Metformin may inhibit the proliferation of HBZY-1 by increasing the expression of p-AMPK in mesangial cells and decreasing the expression of TGF-β1.