摘要
为了探讨Sema4D基因对绝经后骨质疏松患者骨髓间充质干细胞(BMMSCs)成骨及成脂肪分化能力的影响,设计3条Sema4D基因的siRNAs,分别通过显微观察和蛋白质免疫印迹(Western blot)鉴定其对BMMSCs生长形态以及对Sema4D蛋白表达的影响.进一步利用实时荧光定量PCR(real-time PCR)和Western blot检测Sema4D基因沉默对成骨指标抗碱性磷酸酶(ALP)、骨钙素(BGP)、胶原蛋白Ⅰ(CollagenⅠ)和成脂肪分化指标脂蛋白酶(LPL)、脂肪细胞结合蛋白2(AP-2)、瘦素(Leptin)表达的影响以及对骨保护素和可溶性细胞核因子-κB受体活化因子及配基(OPG/RANKL/RANK)系统的影响.结果显示Sema4D基因沉默后,绝经后骨质疏松患者BMMSCs中ALP、BGP、CollagenⅠ的表达水平明显升高,LPL、AP-2、Leptin的表达水平也明显升高;OPG的表达水平明显升高,而RANKL和RANK的表达水平明显下降.由上述结果可知,Sema4D基因可以抑制BMMSCs成骨分化和成脂肪分化相关基因的表达,可能通过下调OPG/RANKL/RANK系统相关基因的表达来实现.
To investigate the effect of Sema4D gene silence on osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells （BMMSCs） in patients with postmenopausal osteoporosis,three siRNA,s of Sema4D gene were designed and used to evaluate its effect on growth and morphology of BMMSCs.The expression of ALP, BGP, Collagen I , LPL, AP-2, Leptin and OPG/ RANKL/RANK were analyzed using real-time PCR and Western blot.The results showed that the expression of ALP,BGP,Collagen I ,LPL,AP-2 and Leptin in BMMSCs in patients with postmenopausal osteoporosis significantly increased after Sema4D gene silence. Furthermore, the expression of OPG significantly increased, while the expression of RANKL and RANK significantly decreased.h is suggested that Sema4D can inhibit osteogenic and adipogenic differentiation of BMMSCs in patients with postmenopausal osteoporosis,possibly by regulating OPG/RANKL/RANK signaling pathway, which is expected to be the target of prevention and treatment of postmenopausal osteoporosis.
作者
张韬
陈冬冬
翁艳
冯尔宥
张怡元
ZHANG Tao CHEN Dongdong WENG Yan FENG Eryou ZHANG Yiyuan(Department of Orthopedics Institute,Fuzhou Second Hospital Department of Orthopedics, Fuzhou Second Hospital, Fuzhou 350007,China)
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2017年第2期173-177,共5页
Journal of Xiamen University：Natural Science
基金
福建省医学创新课题(2014-CXB-23)
