摘要
目的探讨生长分化因子11(GDF11)对糖尿病小鼠来源的脂肪间充质干细胞(ADSCs)成骨分化的影响及信号机制。方法分离、培养糖尿病小鼠ADSCs。流式细胞术检测GDF11对ADSCs增殖的影响。成骨诱导培养后,检测碱性磷酸酶(ALP)活性,实时荧光定量PCR(qPCR)检测成骨相关基因的表达,Western blot检测PI3K、Akt的磷酸化水平。结果GDF11不影响ADSCs增殖,呈浓度依赖性地增强ALP活性,且具有饱和性;显著上调Runx2[(2.41±0.19)vs.(1.00±0.11)]、Osx[(1.41±0.21)vs.(1.00±0.10)]和ALP[(1.42±0.20)vs.(1.00±0.09)]的表达(P<0.05);明显提高PI3K[(2.84±0.19)vs.(1.00±0.11)]和Akt[(4.58±0.20)vs.(1.00±0.19)]的磷酸化水平(P<0.05)。而且,GDF11促进糖尿病小鼠ADSC s成骨分化的作用被PI3K抑制剂LY294002部分减弱。结论GDF11促进糖尿病小鼠ADSCs骨向分化,其作用机制可能与激活PI3K/Akt信号通路有关。
Objective To investigate the effects of growth differentiation factor 11(GDF11)on osteoblast differentiation of adipose derived stem cells(ADSCs)from diabetic mice and related signaling mechanism.Methods ADSCs from diabetic mice were isolated and cultured.The effect of GDF11 on the proliferation of ADSCs was detected by flow cytometry.After cultured in osteogenic differentiation medium,ALP activity was detected,qPCR was used to evaluate the expression of osteoblast genes,and phosphorylation levels of PI3K and Akt were detected by western blot.Results Cell proliferation was less affected by GDF11 intervation,promoted ALP activity in a concentration-dependent but saturated manner,significantly upregulated mRNA expression of Runx2[(2.41±0.19)vs.(1.00±0.11)],Osx[(1.41±0.21)vs.(1.00±0.10)]and ALP[(1.42±0.20)vs.(1.00±0.09)](P<0.05),and significantly increased phosphorylation levels of PI3K[(2.84±0.19)vs.(1.00±0.11)]and Akt[(4.58±0.20)vs.(1.00±0.19)](P<0.05).Moreover,the effects of GDF11 on promoting osteoblast differentiation of ADSCs from diabetic mice was partially weakened by the PI3K inhibitor LY294002.Conclusions GDF11 promotes osteoblast differentiation of ADSCs from diabetic mice,and its mechanism may be related with the activation of PI3K/Akt signaling pathway.
作者
龚闽
朱彪
李欣
梅美
党瑞杰
陈晔
GONG Min;ZHU Biao;LI Xin;MEI Mei;DANG Ruijie;CHEN Ye(Department of Stomatology,Yuquan Hospital of Tsinghua University,Beijing 100040,China;PLA Medical School,Beijing 100853,China;Department of Stomatology,Beijing Shijingshan Hospital,Beijing 100043,China)
出处
《武警医学》
CAS
2024年第1期25-29,共5页
Medical Journal of the Chinese People's Armed Police Force
基金
中国博士后科学基金(2019T120980)
