艾塞那肽通过p38MAPK通路对成骨细胞促增殖和抗凋亡的作用研究

Study on the effects of exenatide on proliferation and anti-apoptosis of osteoblasts via p38MAPK pathway

目的探讨艾塞那肽通过p38MAPK通路对成骨细胞促增殖和抗凋亡的作用机制。方法将小鼠成骨细胞MC3T3-E1 Subclone 14分为对照组、艾塞那肽组、棕榈酸钠组、棕榈酸钠组+艾塞那肽组,分别使用相应试剂培养48 h。通过CCK-8和流式细胞术检测细胞活力与凋亡。通过Western blot检测细胞中P38MAPK、Cyclin D1、Caspase-3的蛋白水平。结果艾塞那肽对正常细胞活力无显著影响(P>0.05),棕榈酸钠组的细胞活力显著低于对照组(P<0.01),艾塞那肽+棕榈酸钠组的细胞活力显著高于棕榈酸钠组(P<0.01);艾塞那肽对正常细胞凋亡无显著影响(P>0.05),棕榈酸钠组的细胞凋亡率显著高于对照组(P<0.01),艾塞那肽+棕榈酸钠组的细胞凋亡率显著低于棕榈酸钠组(P<0.01);艾塞那肽对正常细胞中P38MAPK、Cyclin D1和Caspase-3蛋白水平无明显影响(P>0.05),棕榈酸钠组的P38MAPK和Caspase-3显著高于对照组而Cyclin D1显著低于对照组(P<0.01),艾塞那肽+棕榈酸钠组的P38MAPK和Caspase-3显著低于棕榈酸钠组而Cyclin D1显著高于棕榈酸钠组(P<0.01)。结论艾塞那肽可以通过p38MAPK通路,减少Caspase-3蛋白并上调Cyclin D1蛋白的表达,抑制高脂环境下成骨细胞的凋亡并提高细胞活力。

Objective To explore the mechanism of action of exenatide on proliferation and anti-apoptosis of osteoblasts through p38MAPK pathway.Methods Mouse osteoblast MC3T3-E1 Subclone 14 was divided into control group,exenatide group,sodium palmitate group,sodium palmitate group and exenatide group,and cultured for 48 h with corresponding reagents.Cell viability and apoptosis were detected by CCK-8 and flow cytometry.Western blot was used to detect the protein levels of P38MAPK,Cyclin D1 and Caspase-3 in cells.Results Exenatide had no significant effect on normal cell viability(P>0.05).The cell viability of the sodium palmitate group was significantly lower than that of the control group(P<0.01).The cell viability of the exenatide+palmitate group was significantly higher than that of the sodium palmitate group(P<0.01).Exenatide had no significant effect on normal cell apoptosis(P>0.05).The apoptosis rate of the sodium palmitate group was significantly higher than that of the control group(P<0.01).The apoptosis rate of the exenatide+palmitate group was significantly lower than that of the sodium palmitate group(P<0.01).Exenatide had no significant effect on normal cells(P>0.05).P38MAPK and Caspase-3 in the sodium palmitate group were significantly higher than those in the control group,and Cyclin D1 was significantly lower than the control group(P<0.01).P38MAPK and Caspase-3 in the exenatide+palmitate group were significantly lower than those in the sodium palmitate group and Cyclin D1 was significantly higher than the sodium palmitate group(P<0.01).Conclusion Exenatide can reduce Caspase-3 protein and up-regulate the expression of Cyclin D1 protein through p38MAPK pathway,inhibit osteoblast apoptosis and increase cell viability in high-fat environment.