靶向抑制PI3K对人体外周血破骨细胞分化p38/c-Fos信号通路调控的研究  

The regulation of p38/c-Fos signal pathway by targeting inhibition of PI3K in human peripheral osteoclasts

作　　者：王想福[1] 孙凤歧[1] 石瑞芳[1] 王兴盛[2] 叶丙霖[2] 范有福[1] 杨峰[1] 

机构地区：[1]甘肃省中医院,兰州730050 [2]甘肃省中医药研究院,兰州730050

出　　处：《中国骨质疏松杂志》2016年第1期49-52,共4页Chinese Journal of Osteoporosis

基　　金：陇原青年创新人才扶持计划

摘　　要：目的探讨靶向抑制磷脂酰肌醇3-激酶(PI3K)调控p38/c-Fos信号通路对破骨细胞分化的影响。方法采用人体外周血分离单个核细胞,并诱导形成破骨细胞,干预组应用靶向抑制PI3K的LY294002对细胞进行处理,通过细胞形态学观察并检测细胞活性,再分别采用RT-PCR和Western-Blot两种方法考察p38MAPK通路及下游转录因子c-fos基因和蛋白的表达水平。结果模型组与干预组细胞形态学观察均可见诱导培养的破骨细胞形态明显;与模型组比较,干预组破骨细胞样细胞的形成减少,活性减弱,具有统计学意义(P<0.05);RT-PCR结果均显示,干预组p38MAPK通路下游转录因子c-fos表达水平较对照组下降,有显著性差异(P<0.01);Western-Blot检测结果显示干预组c-fos蛋白表达水平较对照组降低,具有统计学意义(P<0.05)。结论靶向抑制PI3K在体外细胞培养中可抑制破骨细胞形态,并可通过下调p38MAPK通路中c-fos基因和蛋白水平的表达,从而减弱破骨细胞吸收功能。Objective To investigate the effect of targeting inhibition of PI3K on the regulation of p38/c-Fos signaling pathway in osteoclast differentiation. Methods Mononuclear cells were isolated from human peripheral blood and were induced to osteoclasts. Cells in the treatment group were treated with PI3K targeting inhibitor LY294002. Cell morphology was observed and cell proliferation was measured. The p38MAPK pathway and the gene and protein expression level of transcription factor c-los were detected using RT-PCR and Western blotting. Results The cells showed osteoclast morphology both in the model group and intervention group, The osteoclast-like cells significantly reduced and the cell viability decreased in the intervention group comparing to those in the model group, The RT-PCR result showed that c-fos expression level in the intervention group was significantly lower than that in control group （P 〈 0.01 ）. The c-los protein expression level in the intervention group was significantly lower than that in control group （P 〈 0.05）. Conclusion Targeting inhibition of PI3K can inhibit osteoclast formation in vitro. It reduces the function of osteoclasts by down-regulation of c-los gene and protein in p38MAPK pathway.

关 键 词：靶向抑制 骨质疏松 破骨细胞 信号通路 

分 类 号：R318[医药卫生—生物医学工程]