二甲双胍调节PI3K/AKT通路对高糖诱导牙周膜成纤维细胞凋亡的影响研究

doi:10.13701/j.cnki.kqyxyj.2020.12.006

摘要/Abstract

摘要： 目的:探讨二甲双胍对高糖诱导的牙周膜成纤维细胞(PDLF)凋亡及对磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)通路的影响。方法:体外培养人PDLF细胞,设置NG组(5 mmol/L D-葡萄糖)、HG组(30 mmol/L D-葡萄糖)、NG+Met组(5 mmol/L D-葡萄糖+2 mmol/L二甲双胍)、HG+Met组(30 mmol/L D-葡萄糖+2 mmol/L二甲双胍)和HG+Met+Mil组(30 mmol/L D-葡萄糖+2 mmol/L二甲双胍+50 nmol/L Miltefosine),培养48 h。采用CCK-8法检测PDLF细胞增殖情况;采用流式细胞仪检测PDLF细胞凋亡情况和细胞周期分布情况;采用Western blot法检测PDLF细胞中p-PI3K、PI3K、p-AKT、AKT蛋白表达情况。结果:与NG组相比,HG组、HG+Met组、HG+Met+Mil组PDLF细胞A值、S、G2/M期细胞比例、p-PI3K/PI3K、p-AKT/AKT蛋白表达水平显著降低(P＜0.05),凋亡率、G0/G1期PDLF细胞比例显著升高(P＜0.05);与HG组相比,NG+Met组、HG+Met组、HG+Met+Mil组PDLF细胞A值、S、G2/M期细胞比例、p-PI3K/PI3K、p-AKT/AKT蛋白表达水平显著升高(P＜0.05),凋亡率、G0/G1期PDLF细胞比例显著降低(P＜0.05);与NG+Met组相比,HG+Met组、HG+Met+Mil组PDLF细胞A值、S、G2/M期细胞比例、p-PI3K/PI3K、p-AKT/AKT蛋白表达水平显著降低(P＜0.05),凋亡率、G0/G1期PDLF细胞比例显著升高(P＜0.05);与HG+Met组相比,HG+Met+Mil组PDLF细胞A值、S、G2/M期细胞比例、p-PI3K/PI3K、p-AKT/AKT蛋白表达水平显著降低(P＜0.05),凋亡率、G0/G1期PDLF细胞比例显著升高(P＜0.05)。结论:二甲双胍可抵抗高糖诱导的PDLF凋亡,可能是通过激活PI3K/AKT通路实现的。

关键词: 二甲双胍, 磷脂酰肌醇3-激酶/蛋白激酶B通路, 高糖, 牙周膜成纤维细胞, 凋亡

Abstract: Objective: To investigate the effects of metformin on the apoptosis of periodontal ligament fibroblasts (PDLF) induced by high glucose and on the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway. Methods: PDLF cells were cultured in vitro, and the cells were divided into NG group (5 mmol/L D-glucose), HG group (30 mmol/L D-glucose), NG + Met group (5 mmol/L D-glucose + 2 mmol/L metformin), HG + Met group (30 mmol/L D-glucose + 2 mmol/L metformin), and HG + Met + Mil group (30 mmol/L D-glucose + 2 mmol/L metformin + 50 nmol/L miltefosine). The cells were cultured for 48 hours. CCK-8 method was used to detect the proliferation of PDLF cell. The apoptosis and cell cycle distribution of PDLF were detected by flow cytometry. The expressions of p-PI3K, PI3K, p-AKT, and AKT in PDLF cells were detected by Western blotting. Results: Compared with NG group, the OD value of PDLF cells, proportions of PDLF cells in S and G2/M phases, protein expression levels of p-PI3K/PI3K and p-AKT/AKT in HG group, HG+Met group, and HG+Met+Mil group were significantly lower (P<0.05), and the apoptosis rate and proportion of PDLF cells in G0/G1 phase were significantly higher (P<0.05). Compared with HG group, the OD value of PDLF cells, proportions of PDLF cells in S and G2/M phases, and protein expression levels of p-PI3K/PI3K and p-AKT/AKT in NG+Met group, HG+Met group, and HG+Met+Mil group were significantly higher (P<0.05), and the apoptosis rate and proportion of PDLF cells in G0/G1 phase were significantly lower (P<0.05). Compared with NG+Met, the OD value of PDLF cells, proportions of PDLF cells in S and G2/M phases, and protein expression levels of p-PI3K/PI3K and p-AKT/AKT in HG+Met group and HG+Met+Mil group were significantly lower (P<0.05), and the apoptosis rate and the proportion of PDLF cells in G0/G1 phase were significantly higher (P<0.05). Compared with HG + Met group, the OD value of PDLF cells, proportions of PDLF cells in S and G2/M phases, protein expression levels of p-PI3K/PI3K and p-AKT/AKT in HG + Met + Mil group were significantly lower (P<0.05), and the apoptosis rate and proportion of PDLF cells in G0/G1 phase were significantly higher (P<0.05). Conclusion: Metformin can resist PDLF apoptosis induced by high glucose, which may be achieved by activating PI3K/AKT pathway.

Key words: Metformin, phosphatidyl inositol 3-kinase/protein kinase B pathway, high glucose, periodontal ligament fibroblasts, apoptosis

梁德凤, 周鑫才, 吴芸菲. 二甲双胍调节PI3K/AKT通路对高糖诱导牙周膜成纤维细胞凋亡的影响研究[J]. 口腔医学研究, 2020, 36(12): 1103-1107.

LIANG Defeng, ZHOU Xincai, WU Yunfei. Effect of Metformin on Apoptosis of Periodontal Ligament Fibroblasts Induced by High Glucose[J]. Journal of Oral Science Research, 2020, 36(12): 1103-1107.

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