CD147 对体外破骨细胞分化成熟的影响.docVIP

CD147对体外破骨细胞分化成熟的影响 廖通权，刘玉刚，胡 旭，张 莹，余世明，初同伟 （400037 重庆，第三军医大学新桥医院骨科） [摘要] 目的 建立破骨细胞体外分化成熟的模型，研究CD147对破骨细胞分化成熟的影响。方法 从外周血分离单个核细胞，贴壁培养，诱导破骨细胞生成。实验分为对照组、诱导组（30 ng/ml RANKL+25 ng/ml M-CSF）及阻断组（30 ng/ml RANKL+25 ng/ml M-CSF +10 μg/ml CD147Ab），通过免疫荧光染色和流式细胞术检测CD147分子在培养24、48 h破骨前体细胞上的定位表达和表达强度，TRAP染色观察破骨细胞的成熟，Real-time PCR检测培养24、48 h破骨前体细胞CD147 mRNA、TRAP mRNA的相对表达。结果 ①CD147分子在破骨前体细胞膜上表达；平均荧光强度：诱导48 h组（178.47±4.59）＞诱导24 h组（171.18±3.46）＞对照组（159.55±3.65）（P＜0.05），阻断24 h组显著降低（P＜0.01）。②阻断组TRAP染色阳性细胞（细胞核≥3）数显著少于诱导组（P＜0.01），且破骨细胞体积小。③CD147 mRNA相对表达量：诱导48 h组（1.772+0.326）＞诱导24 h组（1.393±0.248）＞对照组、诱导48 h组（1.772+0.326）＞阻断48 h组（1.242+0.355）、诱导24 h组（1.393±0.248）＞阻断24 h组（0.673±0.175）（P＜0.05）；TRAP mRNA相对表达量：诱导48 h组（2.608±0.906）＞诱导24 h组（1.810±0.398）＞阻断48 h组（0.900±0.128）＞阻断24 h组（0.784±0.265）（P＜0.05）；且TRAP mRNA相对表达量与CD147 mRNA相对表达量呈正相关（R=0.833，P＜0.01）。结论 CD147分子在破骨前体细胞膜上表达且参与破骨细胞分化成熟，抑制CD147分子的表达破骨细胞出现成熟障碍。 [关键词] 破骨细胞；CD147；分化成熟 [中图法分类号] [文献标志码] A Effect of CD147 on the differentiation and maturation of osteoclasts in vitro Liao Tongquan, Liu Yugang, Hu Xu,Zhang Yin, Yu Shiming,Chu Tongwei (Department of Orthopedics, Xinqiao Hospital, Third Military Medical University,Chongqing，400037，China) [Abstract] Objective To establish the model of differentiation and maturation of osteoclasts in vitro and to study the effect of CD147 on the differentiation and maturation of osteoclasts. Methods The mononuclear cells were isolated from peripheral blood and cultured adherently to induce osteoclastogenesis. There are control group, induction group (30ng/ml RANKL+25ng/ml M-CSF) and blocking group (30ng/ml RANKL+25ng/ml M-CSF +10μg/ml CD147Ab).We detected the position of CD147 on osteoclast precursors by Immunofluorescence staining and the quantity of fluorescence intensity of osteoclast precursors which were cultured 24h or 48h by Flow cytometry, and observed the maturing of osteoclasts by TRAP staining. We also detected the expression of CD147mRNA and TRAPmRNA of osteoclast precursors which