鹿茸多肽对人骨髓间充质干细胞BMP-2和Runx2表达的影响  

Influence of velvet antler polypeptides in expressions of BMP-2 and Runx2 in human bone marrow mesenchymal stem cells

作　　者：张洪长[1] 张莹[2] 刘明昕[3] 潘志[1] 律广富[1] 陈港[1] 姜鸿远 郭骏骐[4] 

机构地区：[1]长春中医药大学中医药与生物工程研究开发中心药理研究室,吉林长春130117 [2]吉林大学药学院药理学教研室,吉林长春130021 [3]长春中医药大学图书馆参考咨询部,吉林长春130117 [4]长春中医药大学科技处,吉林长春130117

出　　处：《吉林大学学报（医学版）》2015年第3期491-495,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81374025);吉林省科技厅科技发展计划项目资助课题(20140414051GH)

摘　　要：目的:探讨鹿茸多肽(VAP)对人骨髓间充质干细胞(hBMSCs)中骨形态发生蛋白2(BMP-2)和骨特异性转录因子2(Runx2)基因和蛋白表达的影响,阐明其对骨合成及hBMSCs向成骨细胞方向分化的作用机制。方法:体外培养的hBMSCs分为对照组和5、10、15、20g·L-1 VAP组,通过检测各组hBMSCs中碱性磷酸酶(ALP)活性,确定体外用药最佳药物浓度;CCK-8法检测对照组和10g·L-1 VAP组hBMSCs增殖情况;荧光定量PCR法与Western blotting法测定对照组和10g·L-1 VAP组hBMSCs中BMP-2和Runx2基因与蛋白的表达。结果:VAP各组细胞中ALP活性均高于对照组,其中以10g·L-1 VAP组细胞中ALP活性最高(P<0.01);CCK-8法,10g·L-1 VAP组hBMSCs增殖率明显高于对照组(P<0.05),VAP诱导培养第9天hBMSCs增殖率最高,且进入平台期,之后hBMSCs的增殖率开始下降;荧光定量PCR和Western blotting检测,10g·L-1 VAP组hBMSCs中BMP-2和Runx2基因及蛋白表达水平明显高于对照组,差异有统计学意义(P<0.05)。结论:VAP能够提高hBMSCs中ALP活性,促进hBMSCs增殖分化,上调hBMSCs中BMP-2及其下游Runx2基因和蛋白的表达,这可能是VAP对hBMSCs骨形成及骨代谢影响的分子调控机制之一。Objective To investigate the influence of velvet antler polypeptides（VAP）in the expressions of bone morphogenetic protein-2（BMP-2）and runt-related transcription factor 2（Runx2）gene and protein in human bone marrow mesenchymalstem cells（hBMSCs）,and to clarify the mechanism of VAP in bone synthesis and bone differentiation direction.Methods The cultured hBMSCs were divided into control group and 5,10,15,20 g ·L^-1 VAP groups.By detecting the alkaline phosphatase（ALP）activities in hBMSCs in various groups,the optimal concentration of the drug in vitro was selected;CCK-8assay was used to detect the proliferation of hBMSCs in control group and 10g·L^-1 VAP group;the BMP-2and Runx2 gene and protein expressions in control group and 10g·L^-1 VAP group were determined with fluorescence quantitative PCR and Western blotting method.Results The ALP activities in VAP groups were higher than that in control group,and the ALP activity in10 g ·L^-1 VAP group was the highest（P〈0.01）.The CCK-8assay results showed that the proliferation rate of the hBMSCs in 10 g ·L^-1 VAP group was higher than that in control group（P〈0.05）,the VAP-induced proliferation of cultured hBMSCs was the highest at the 9th day,then went into the plateau and began to decline;the quantitative PCR and Western blotting results showed that the expression levels BMP-2and Runx2 gene and protein in the hBMSCs in 10 g ·L^-1 VAP group were significantly higher than those in control group,and the differences were statistically significant（P〈0.05）.Conclusion VAP can improve the ALP activity in hBMSCs and promote the proliferation and differentiation of hBMSCs,and increase the expressions of BMP-2gene and its Runx2 gene and protein,which may be one of the molecular regulation mechanisms of VAP for bone formation and bone metabolism of hBMSCs.

关 键 词：鹿茸多肽 人骨髓间充质干细胞 骨形态发生蛋白2 骨特异性转录因子2 

分 类 号：R961[医药卫生—药理学]