人破骨细胞分化中重组结核杆菌热休克蛋白10的影响  

Recombinant mycobacterium tuberculosis heat shock protein 10 in human osteoclast differentiation

作　　者：张元豫 郭永荣 刘霞[2] 李坤 

机构地区：[1]新疆自治区人民医院骨科,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

出　　处：《中国组织工程研究》2014年第38期6116-6122,共7页Chinese Journal of Tissue Engineering Research

基　　金：新疆自然科学基金(2011211A054)~~

摘　　要：背景:体外小鼠颅盖骨实验已证实了其结核杆菌热休克蛋白10的破骨效应。目的:在诱导破骨细胞分化的体外细胞培养系统中,研究重组结核杆菌热休克蛋白10对破骨细胞分化的影响及相关机制。方法:选用人巨噬细胞集落刺激因子依赖性附着性血液单个核细胞,实验分为:核因子κB受体活化因子配体+重组结核杆菌热休克蛋白10组、核因子κB受体活化因子配体组、重组结核杆菌热休克蛋白10组(1 mg/L)、阴性对照组(完全培养基),核因子κB受体活化因子配体+重组结核杆菌热休克蛋白10组、重组结核杆菌热休克蛋白10组中重组结核杆菌热休克蛋白10质量浓度为1 mg/L,在含有巨噬细胞集落刺激因子的a-MEM培养液重悬单核细胞,各组培养7,14,21 d后,观察所形成的抗酒石酸酸性磷酸酶阳性染色多核细胞的形态、数目、骨吸收面积;各组NFATc1、c-Fos基因及蛋白表达情况。结果与结论:阴性对照组无抗酒石酸酸性磷酸酶阳性多核破骨细胞分化生成,其余各组均有抗酒石酸酸性磷酸酶阳性多核破骨细胞分化生成,并在小牛骨磨片上形成吸收陷窝;重组结核杆菌热休克蛋白10组形成的破骨细胞分化细胞数目、吸收陷窝数目及陷窝面积显著低于核因子κB受体活化因子配体+重组结核杆菌热休克蛋白10组;阴性对照组NFATc1、c-Fos基因表达水平显著低于核因子κB受体活化因子配体+重组结核杆菌热休克蛋白10组和重组结核杆菌热休克蛋白10组,而重组结核杆菌热休克蛋白10组表达NFATc1、c-Fos蛋白,显著低于核因子κB受体活化因子配体+重组结核杆菌热休克蛋白10组。提示结核杆菌热休克蛋白10参与破骨细胞分化的形成,可能与诱导相关基因NFATc1、c-Fos表达上调有关。BACKGROUND： The mycobacterium tuberculosis heat shock protein 10 exerts effects on the osteoclasts by in vitro mouse cranium experiment, OBJECTIVE： To investigate the effect and mechanism of recombinant mycobacterium !uberculosis heat shock protein 10 （CPN10） on the differentiation of osteoclasts in the in vitro culture system that induces osteoclast differentiation.METHODS： Human macrophage colony-stimulating factor-dependent adhesive blood mononuclear cells were divided into four groups： receptor activator for nuclear factor-KB ligand （RANKL）＋CPN10 （1 mg/L）, RANKL, CPN10 （1 mg/L）, and negative control （complete culture medium）. Monocytes were resuspended in a-MEM medium containing macrophage colony-stimulating factor, and were cultured in each group for 7, 14, 21 days. The morphology, quantity and bone resorption area of osteoclasts were examined by tartrate-resistant acid phosphatase （TRAP） staining. The expressions of NFATcl and c-Fos gene and protein were also detected. RESULTS AND CONCLUSION： In negative control group, no TRAP-positive multinucleated osteoclasts generated, while in the other groups, TRAP-positive multinucleated osteoclasts differentiated and formed the lacunae in the small bone grinding. The number of osteoclasts formation and resorption in CPN10 group were significantly lower than that in RANKL＋CPN10 group. The expression of NFATcl and c-Fos in the negative control group C was significantly lower than that of RANKL＋CPN10 group and CPN10 group. However, CPN10 expressed NFATcl and c-Fos protein, which was significantly lower than RANKL＋CPN10 group. CPN10 is involved in the formation of osteoclasts, and the mechanism is related with the upregulation of NFATcl, c-Fos expression.

关 键 词：破骨细胞 NFATC转录因子类 基因 FOS 组织构建 骨组织工程 重组结核杆菌热休克蛋白10 单个核细胞 NFATc1 c-Fos 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]