颗粒蛋白前体促进去卵巢小鼠的骨质疏松  

Progranulin aggravates postmenopausal osteoporosis in ovariectomized mice

作　　者：赵安平 朱卫文[1] 郭民康 杨武 张健[1] ZHAO Anping;ZHU Wei wen;GUO Minkang;YANG Wu;ZHANG Jian(Department of Orthopedics,First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学附属第一医院骨科

出　　处：《细胞与分子免疫学杂志》2019年第8期714-720,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：重庆市卫生计生委医学科研项目重点项目(2017ZDXM006);重庆市科学技术委员会基础与前沿科技项目(20150121)

摘　　要：目的探讨颗粒蛋白前体(PGRN)对去卵巢小鼠骨质疏松的影响。方法建立野生型和PGRN敲除(PGRN-/-)小鼠卵巢切除骨质疏松模型,采用微型计算机断层扫描(Micro-CT)和三维重建获得小鼠骨组织微观结构并进行骨小梁数据分析,HE染色观察骨组织形态,抗酒石酸酸性磷酸酶(TRAP)染色观察骨组织中破骨细胞数量,免疫组织化学染色法检测骨组织核因子κB受体激活蛋白配体(RANKL)、肿瘤坏死因子α(TNF-α)、P65表达,荧光定量PCR检测骨组织TRAP mRNA水平,Western blot法检测骨组织基质金属蛋白酶9(MMP9)、MMP14、P65蛋白水平。结果与野生型小鼠相比,PGRN-/-组小鼠骨密度(BMD)、骨体积分数(BV/TV)、骨小梁数目(Tb.N)及骨小梁厚度(Tb.Th)均显著增加,骨小梁空间距(Tb.S)显著降低;在PGRN-/-小鼠中观察到骨质疏松程度更轻,破骨细胞数量更少,骨组织RANKL、TNF-α、P65蛋白,TRAP mRNA,MMP9、MMP14表达均低于野生型小鼠。结论PGRN促进卵巢切除小鼠骨质疏松。Objective To investigate the effect of progranulin(PGRN) on osteoporosis in ovariectomized mice. MethodsPGRN-knockout(PGRN-/-) and wild-type mice were ovariectomized to induce postmenopausal osteoporosis models. Next, the bone tissues in all mice were scanned by Micro-CT and three-dimensional reconstruction was performed to detect the micro-structure, followed by trabecula data analysis. The morphology and osteoclasts in the bone tissues of PGRN-/- and wild-type mice were observed by HE staining and TRAP staining, respectively. The expression of receptor activator for nuclear factor-κB ligand(RANKL), tumor necrosis factor α(TNF-α) and P65 were detected by immunohistochemistry. The expression of TRAP mRNA in the mice was measured using fluorescence quantitative PCR and the protein expression of MMP9, MMP14, P65 was detected by Western blot analysis. Results Bone mineral density(BMD), bone volume fraction(BV/TV), trabecular number(Tb.N) and trabecular thickness(Tb.Th) in the PGRN-/- group were significantly higher than those in the wild-type group, while the trabecular separation(Tb.S) in the PGRN-/- group was in the contrary. The degree of osteoporosis was less severe and number of osteoclasts in the PGRN-/- mice were reduced, likewise, RANKL, TNF-α, MMP9, MMP14 and P65 as well as TRAP mRNA were down-regulated in the PGRN-/- group compared with the wild-type group. Conclusion PGRN aggravates the postmenopausal osteoporosis in ovariectomized mice.

关 键 词：颗粒蛋白前体(PGRN) 卵巢切除 绝经后骨质疏松 破骨细胞 雌激素 

分 类 号：R392-33[医药卫生—免疫学] R713.6[医药卫生—基础医学]