目的 探讨生长分化因子11(GDF11)对糖尿病小鼠骨髓间充质干细胞(BMSCs)细胞凋亡的影响及其信号机制。方法 复苏BMSCs并将其分为4组:正常小鼠BMSCs组(Nor),糖尿病小鼠BMSCs组(DB),GDF11干预组(DB+GDF11;GDF11),抑制剂组(DB+GDF11+ LY294002;LY)。各组细胞培养3 d后,台盼蓝染色观察细胞凋亡率。Western blot检测凋亡相关蛋白Bax、Bcl-2和磷酸化(p)-PI3K、p-Akt的表达水平。结果 与正常小鼠BMSCs相比,糖尿病小鼠BMSCs细胞凋亡率显著升高[(36.2±3.3)% vs. (3.1±1.1)%,P＜0.05];GDF11可降低糖尿病小鼠BMSCs细胞凋亡率[(18.9±1.9)% vs. (36.2±3.3)%,P＜0.05],提高Bcl-2/Bax的蛋白表达比[(1.63±0.10) vs. (0.26±0.09);P＜0.05],同时激活PI3K[p-PI3K/PI3K:(0.98±0.08) vs. (0.42±0.11);P＜0.05]/Akt[p-Akt/Akt:(0.94±0.10) vs. (0.32±0.15);P＜0.05]信号通路。而且,当抑制PI3K/Akt信号通路后,GDF11抑制糖尿病小鼠BMSCs凋亡的作用明显减弱[细胞凋亡率:(41.1±3.9)% vs. (18.9±1.9)%,P＜0.05]。结论 GDF11可通过激活PI3K/Ak信号通路抑制糖尿病小鼠BMSCs细胞凋亡。
Abstract
Objective To investigate the effects of GDF11 on apoptosis of BMSCs from diabetic mice and the related mechanism.Methods BMSCs of normal mice (Nor),diabetic mice (DB),DB with GDF11 intervention (GDF11),and DB+GDF11 plus LY294002 (LY) were cultured for 3 days.Trypan blue staining was used to calculate the apoptotic rate,while western blot was used to investigate protein levels of Bax,Bcl-2,phosphorylated (p)-PI3K and p-Akt.Results Compared with Nor,the apoptotic rate of DB was increased [(36.2±3.3)% vs. (3.1±1.1)%,P＜0.05],but GDF11 intervention could reduce the apoptotic rate [(18.9±1.9)% vs. (36.2±3.3)%,P＜0.05],increase Bcl-2/Bax expression[(1.63±0.10) vs. (0.26±0.09);P＜0.05],and activate the PI3K[p-PI3K/PI3K:(0.98±0.08) vs. (0.42±0.11);P＜0.05]/Akt[p-Akt/Akt:(0.94±0.10) vs. (0.32±0.15);P＜0.05]signaling pathway.However,when inhibiting PI3K/Akt signaling pathway,GDF11 had its inhibitory effect on cell apoptosis blunted[apoptotic rate:(41.1±3.9)% vs. (18.9±1.9)%,P＜0.05].Conclusions GDF11 can inhibit the apoptosis of BMSCs from diabetic mice by activating the PI3K/Akt signaling pathway.
关键词
生长分化因子11 /
糖尿病 /
骨髓间充质干细胞 /
细胞凋亡
Key words
DGF11 /
diabetes /
bone-marrow mesenchymal stem cells (BMSCs) /
cell apoptosis
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基金
中国博士后科学基金资助项目(2019T120980)
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