生长分化因子11激活PI3K/Akt信号途径促进糖尿病小鼠ADSCs体外矿化的研究
Promotion of GDF11 on osteoblast differentiation of ADSCs derived from diabetic mice by activating PI3K/Akt signaling pathway
龚闽 1朱彪 2李欣 3梅美 1党瑞杰 2陈晔1
作者信息
- 1. 100040 北京,清华大学玉泉医院口腔科
- 2. 100853 北京,解放军医学院
- 3. 100043,北京市石景山医院口腔科
- 折叠
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AD-SC 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.
Abstract
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 i-solated 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 signifi-cantly 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 weak-ened 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.
关键词
生长分化因子11/PI3K/Akt信号通路/脂肪间充质干细胞/成骨分化Key words
growth differentiation factor 11(GDF11)/PI3K/Akt signaling pathway/ADSCs/osteoblast differentiation引用本文复制引用
基金项目
中国博士后科学基金(2019T120980)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据