重组结核杆菌热休克蛋白10对人成骨细胞增殖及骨代谢的调节  

Effect of recombinant Mycobacterium tuberculosis heat shock protein 10 on proliferation of human osteoblasts and regulation of bone metabolism

作　　者：张元豫[1] 刘霞[2] 李坤[3] 郭永荣[3] 白靖平[1] 

机构地区：[1]新疆医科大学附属肿瘤医院骨科,新疆维吾尔自治区乌鲁木齐市830000 [2]新疆医科大学第一附属医院病理科,新疆维吾尔自治区乌鲁木齐市830000 [3]新疆维吾尔自治区人民医院骨科,新疆维吾尔自治区乌鲁木齐市830000

出　　处：《中国组织工程研究》2014年第11期1665-1671,共7页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金(2011211A054)~~

摘　　要：背景:结核杆菌热休克蛋白10是引起骨结核骨质溶解和吸收主要因素之一,抑制成骨细胞的增殖。核因子κB受体活化因子配体及骨保护素是影响骨代谢的重要指标。目的:观察重组结核杆菌热休克蛋白10对人成骨细胞增殖、碱性磷酸酶活性和核因子κB受体活化因子配体mRNA及骨保护素mRNA表达的影响。方法:将人骨髓基质干细胞定向诱导分化筛选为成骨细胞,取第3代细胞,用含质量浓度为0.1,1,10 mg/L的重组结核杆菌热休克蛋白10培养液培养,对照组成骨细胞正常培养。结果与结论:MTT实验结果显示,与对照组相比,不同质量浓度重组结核杆菌热休克蛋白10抑制成骨细胞增殖和碱性磷酸酶活性(P<0.05)。RT-PCR实验显示,与对照组相比,不同质量浓度的重组结核杆菌热休克蛋白10均增加成骨细胞中核因子κB受体活化因子配体mRNA表达(P<0.01),降低护骨素mRNA表达(P<0.01),均呈剂量依赖性,质量浓度10 mg/L的重组结核杆菌热休克蛋白10作用最明显(P<0.01)。结果证实,重组结核杆菌热休克蛋白10抑制成骨细胞增殖和碱性磷酸酶活性,通过调节核因子κB受体活化因子配体mRNA及骨保护素mRNA表达而影响骨代谢。BACKGROUND：Mycobacterium tuberculosis heat shock protein 10 （r-Mt cpn10） is one of the main factors that cause bone tuberculosis dissolution and absorption as wel as inhibits the proliferation of osteoblasts. Receptor activator of nuclear factor kappa B ligand and osteoprotegerin are the important factors influencing bone metabolism.OBJECTIVE：To observe the effect of r-Mt cpn10 on human osteoblast proliferation, alkaline phosphatase secretion, expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA. METHODS：Human bone marrow stromal cel s were induced to differentiate into osteoblasts, and osteoblasts at passage 3 were cultured with various concentrations of r-Mt cpn10 （0.1, 1, 10 mg/L）. Osteoblasts cultured without r-Mt CPN10 were assigned as controls.RESULTS AND CONCLUSION：MTT assay results showed that, compared with control group, r-Mt cpn10 at different concentrations inhibited osteoblast proliferation and alkaline phosphatase secretion （P〈0.05）. RT-PCR analysis showed that, r-Mt cpn10 at different concentrations increased receptor activator of nuclear factor-kappa B＆amp;nbsp;ligand mRNA expression （P〈0.01）, and inhibited osteoprotegerin mRNA expression in a concentration-dependent manner （P〈0.01）. 10 mg/L r-Mt cpn10 exhibited the strongest effect （P〈0.01）. The r-Mt cpn10 can inhibit osteoblast proliferation and alkaline phosphatase activity, and it may influence bone metabolism by regulating the expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA.

关 键 词：组织构建 骨组织构建 成骨细胞 细胞增殖 热休克蛋白质类 碱性磷酸酶 新疆维吾尔自治区自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]