Effect of Metformin on Apoptosis of Periodontal Ligament Fibroblasts Induced by High Glucose

doi:10.13701/j.cnki.kqyxyj.2020.12.006

Abstract

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

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.

References

[1] 谢佳颖,尹东青.缺氧对人牙周膜细胞增殖、迁移能力及IGF-1表达的影响[J].重庆医科大学学报,2019,44(5):561-567.
[2] 鲍千红,徐公民,吴建荣,等.2型糖尿病患者糖化血红蛋白水平与并发牙周疾病的关系[J].中国医师杂志,2017,19(3):414-417.
[3] 靳东思奇,廖雁婷,和璐,等.糖尿病前期患者的牙周状况研究[J].中华口腔医学杂志,2019,54(3):157-163.
[4] 郭玲,张云水,詹聃婷,等.RAGE阻断剂FPS-ZM1对高糖环境人牙周膜成纤维细胞增殖、迁移的影响[J].口腔医学研究,2017,33(12):1258-1261.
[5] 庞若宇,关美萍,郑宗基,等.二甲双胍对糖基化终末产物诱导的成纤维细胞凋亡及相关蛋白caspase-3、Bax及Bcl-2表达的影响[J].南方医科大学学报,2015,35(6):898-902.
[6] Zhang XQ, Yao C, Bian WH, et al. Effects of Astragaloside Ⅳ on treatment of breast cancer cells execute possibly through regulation of Nrf2 via PI3K/AKT/mTOR signaling pathway [J]. Food Sci Nutr, 2019, 7(11):3403-3413.
[7] 周志强,冯燕,党海霞.白色波特兰水门汀对人牙周膜成纤维细胞体外增殖的影响[J].中华实用诊断与治疗杂志,2018,32(7):664-666.
[8] Yang YY, Sun XT, Li ZX, et al. Protective effect of angiotensin-(1-7) against hyperglycaemia-induced injury in H9c2 cardiomyoblast cells via the PI3KI/Akt signaling pathway [J]. Int J Mol Med, 2018, 41(3):1283-1292.
[9] Ledesma G, Umpierrez GE, Morley JE, et al. Efficacy and safety of linagliptin to improve glucose control in older people with type 2 diabetes on stable insulin therapy: A randomized trial [J]. Diabetes Obes Metab, 2019, 21(11):2465-2473.
[10] Seubbuk S, Sritanaudomchai H, Kasetsuwan J, et al. High glucose promotes the osteogenic differentiation capability of human periodontal ligament fibroblasts [J]. Mol Med Rep, 2017, 15(5):2788-2794.
[11] 谢颖,李菲,欧阳翔英.糖尿病患者的牙周病[J].中华全科医师杂志,2018,17(6):421-423.
[12] 黄辉,黄炫赓,于大海,等.二甲双胍对实验性糖尿病牙周炎大鼠牙周组织的影响[J].中国现代医生, 2018, 56(4):43-46.
[13] Mohamed TMA, Ang YS, Radzinsky E, et al. Regulation of cell cycle to stimulate adult cardiomyocyte proliferation and cardiac regeneration [J]. Cell, 2018, 173(1):104-116.
[14] Jiang K, Zhi T, Xu W, et al. MicroRNA-1468-5p inhibits glioma cell proliferation and induces cell cycle arrest by targeting RRM1 [J]. Am J Cancer Res, 2017, 7(4):784-800.
[15] Lim W, Park S, Bazer FW, et al. Naringenin-induced apoptotic cell death in prostate cancer cells is mediated via the PI3K/AKT and MAPK signaling pathways [J]. J Cell Biochem, 2017, 118(5):1118-1131.
[16] Kumar D, Haldar S, Gorain M, et al. Epoxyazadiradione suppresses breast tumor growth through mitochondrial depolarization and caspase-dependent apoptosis by targeting PI3K/AKT pathway [J]. BMC Cancer, 2018, 18(1):52-69.
[17] Fang XS, Zhang MH, Zhang XZ, et al. Insulin-like growth factor-1 inhibits the apoptosis of rat gastric smooth muscle cells cultured under high glucose condition through PI3K-Akt-PKC-Ca²⁺ pathway [J]. Biotechnol Biotec Eq, 2019, 33(1):456-464.
[18] 胡涛,韩璐.miR-23b过表达对高糖诱导近端肾小管上皮细胞EMT及PI3K-AKT通路的影响[J].河北医药, 2019, 41(2):190-194.
[19] 梁栋,刘云峰,章毅,等.牛磺酸通过激活PI3K/Akt信号通路抑制高糖诱导的内皮细胞凋亡[J].国际内分泌代谢杂志, 2015, 35(2):73-76.
[20] Liu D, Wu M, Lu Y, et al. Protective effects of 6-Gingerol on vascular endothelial cell injury induced by high glucose via activation of PI3K-AKT-eNOS pathway in human umbilical vein endothelial cells [J]. Biomed Pharmacother, 2017, 93(1):788-795.
[21] 胡晨,李力,卢从华,等.二甲双胍对生物电场下肺癌H1975细胞增殖和定向迁移的影响[J].第三军医大学学报, 2019, 41(6):521-528.
[22] 谢敏,孙英明,孙丽娟,等.二甲双胍联合2-脱氧葡萄糖诱导人宫颈癌Hela细胞凋亡与AKT信号通路相关性的研究[J].医学信息, 2015, 28(40):26-29.