二甲双胍通过IGF-1/PI3K/Akt信号通路对细胞衰老的影响

Effect of metformin on cell senescence through IGF-1/PI3K/Akt signaling pathway

目的探讨二甲双胍(Met)通过IGF-1/PI3K/Akt信号通路对细胞衰老的影响。方法选取2BS细胞分成四组,即对照(NC)组、胰岛素样生长因子-1(IGF-1)组、Met组、Met+IGF-1组。加药处理后进行SA-β-Gal染色检测细胞衰老水平、CCK8试验检测细胞增殖能力、EdU试验检测DNA合成能力以及Western blot检测各组PI3K、p-PI3K、Akt、p-Akt蛋白表达。结果各组SA-β-Gal染色阳性率比较,差异有统计学意义(P<0.05)。Met+IGF-1组SA-β-Gal染色阳性率低于IGF-1组,差异有高度统计学意义(P<0.01)。在相同时间点,Met+IGF-1组OD450值高于IGF-1组,差异有统计学意义(P<0.05)。EdU染色后在370 nm处的染色结果显示,各组OD370值比较,差异有统计学意义(P<0.05);Met+IGF-1组OD370值高于IGF-1组,差异有统计学意义(P<0.05)。Met组p-PI3K、p-Akt蛋白表达量低于其他组,差异有统计学意义(P<0.05)。IGF-1组p-PI3K、p-Akt蛋白表达量高于其他组,差异有统计学意义(P<0.05)。Met+IGF-1组p-PI3K、p-Akt蛋白表达量低于IGF-1组,差异有统计学意义(P<0.05)。结论Met可通过抑制IGF-1/PI3K/Akt信号通路来延缓细胞衰老。

Objective To explore the effect of metformin(Met)on cell senescence through IGF-1/PI3K/Akt signaling pathway.Methods 2BS cells were selected and divided into four groups,namely control(NC)group,insulin-like growth factor-1(IGF-1)group,Met group,Met+IGF-1 group.After drug treatment,SA-β-Gal staining was performed to detect cell senescence,CCK8 test was used to detect cell proliferation ability,EdU test was performed to detect DNA synthesis ability,and Western blot was used to detect the protein expression of PI3K,p-PI3K,Akt,and p-Akt in each group.Results The positive rate of SA-β-Gal staining in each group was compared,and the difference was statistically significant(P<0.05).The positive rate of SA-β-Gal staining in Met+IGF-1 group was lower than that in IGF-1 group,and the difference was highly statistically significant(P<0.01).At the same time point,the OD450 value of the Met+IGF-1 group was higher than that of the IGF-1 group,and the difference was statistically significant(P<0.05).The staining results at 370 nm after EdU staining showed that the OD370 value of each group was compared with statistical significance(P<0.05);the OD370 value of the Met+IGF-1 group was higher than that of the IGF-1 group,and the difference was statistically significant(P<0.05).The expressions of p-PI3K and p-Akt protein in Met group were lower than those in other groups,and the differences were statistically significant(P<0.05).The expression of p-PI3K and p-Akt protein in IGF-1 group were higher than those in other groups,and the differences were statistically significant(P<0.05).The expressions of p-PI3K and p-Akt protein in Met+IGF-1 group were lower than those in IGF-1 group,and the differences were statistically significant(P<0.05).Conclusion Metformin can delay cell senescence by inhibiting IGF-1/PI3K/Akt signaling pathway.