高糖环境下BCA-1通过血管平滑肌细胞对间充质干细胞增殖和凋亡的影响

Effects of BCA-1 on the Proliferation and Apoptosis of Mesenchymal Stem Cells through Vascular Smooth Muscle Cells under High Glucose Environment

目的探讨高糖环境下,B淋巴细胞化学引诱物-1(BCA-1)处理的主动脉平滑肌细胞(ASMC)对间充质干细胞(MSC)增殖和凋亡的影响,并分析其作用机制。方法含30 mmol/L葡萄糖培养基为细胞高糖模型。正常环境培养的小鼠主动脉平滑肌细胞(mASMC)为正常对照组;细胞高糖环境下,mASMC分为高糖对照组,高糖BCA-1组和高糖PI3K抑制剂组。收集各组mASMC条件培养基(CM)培养小鼠骨髓间充质干细胞(mBMSC)。mBMSC分为正常对照组,高糖对照组,高糖CM组、高糖BCA-1CM组和高糖PI3K抑制剂CM组。CCK-8、流式细胞术和Western blot分别检测各组mBMSC增殖吸光度值(A值)、凋亡率和Caspase-8蛋白表达;ELISA检测各组mASMC的Akt、p-Akt蛋白含量。结果与其他组相比,高糖BCA-1CM组mBMSC增殖A值为(1.31±0.15),A值显著增高,差异有统计学意义(P<0.01)。相对于其他各组,高糖BCA-1CM组细胞凋亡率为26.78%,Caspase-8蛋白相对表达量为(0.42±0.04),数据均有明显降低,差异有统计学意义(P<0.01);高糖PI3K抑制剂CM组mBMSC的各项检测指标呈相反变化。相对于高糖对照组,高糖BCA-1组mASMC的Akt、p-Akt蛋白含量分别为(11.85±0.18)μg/mL和(7.69±0.13)μg/mL,数据相比显著高表达,差异有统计学意义(P<0.01);与高糖BCA-1组相比,高糖PI3K抑制剂组mASMC的Akt、p-Akt蛋白含量明显降低,差异有统计学意义(P<0.01)。结论高糖环境下,BCA-1激活mASMC内PI3K-Akt信号通路,上调mBMSC增殖,抑制mBMSC凋亡。

Objective To investigate the effects of B lymphocyte chemoattractant 1(BCA-1)-treated aortic smooth muscle cells(ASMC)on proliferation and apoptosis of mesenchymal stem cells(MSC)in high glucose environment,and to analyze the mechanism.Methods The medium containing 30 mmol/L glucose was used as a cell high-glucose model.Mouse aortic smooth muscle cells(mASMC)cultured in a normal environment served as the normal control group;in a high-glucose environment,mASMCs were divided into a high-glucose control group,a high-glucose BCA-1 group,and a high-glucose PI3K inhibitor group.The mASMC conditioned medium(CM)of each group was collected to culture mouse bone marrow mesenchymal stem cells(mBMSC).mBMSCs were divided into normal control group,high-glucose control group,high-glucose CM group,high-sugar BCA-1CM group and high-sugar PI3K inhibitor CM group.CCK-8,flow cytometry and Western blot were used to detect the proliferation absorbance(A value),apoptosis rate and Caspase-8 protein expression of mBMSC in each group;ELISA was used to detect the Akt and p-Akt protein content of mASMC in each group.Results Compared with other groups,the proliferation A value of mBMSC in the high glucose BCA-1CM group was(1.31±0.15),and the A value was significantly higher,the difference was statistically significant(P<0.01).Compared with other groups,the apoptosis rate of the high glucose BCA-1CM group was 26.78%,the relative expression of Caspase-8 protein was(0.42±0.04),and the data were significantly reduced,the difference was statistically significant(P<0.01);the high glucose PI3K inhibitor CM group indexes of mBMSC showed opposite changes.Compared with the high-glycemic control group,the Akt and p-Akt protein contents of mASMC in the high-glycemic BCA-1 group were(11.85±0.18)μg/mL and(7.69±0.13)μg/mL,respectively.Compared with the data,the expression was significantly higher,the difference was statistically significant(P<0.01);compared with the high-glycemic BCA-1 Compared with the group,the Akt and p-Akt protein contents of mASMC in the high-glycemic PI3K inhibitor group were significantly reduced,the difference was statistically significant(P<0.01).Conclusion Under high glucose environment,BCA-1 activates the PI3K-Akt signaling pathway in mASMC,up-regulates mBMSC proliferation,and inhibits mBMSC apoptosis.