【摘要】：為了探討Sema4D基因?qū)^經(jīng)后骨質(zhì)疏松患者骨髓間充質(zhì)干細胞(BMMSCs)成骨及成脂肪分化能力的影響,設(shè)計3條Sema4D基因的siRNAs,分別通過顯微觀察和蛋白質(zhì)免疫印跡(Western blot)鑒定其對BMMSCs生長形態(tài)以及對Sema4D蛋白表達的影響.進一步利用實時熒光定量PCR(real-time PCR)和Western blot檢測Sema4D基因沉默對成骨指標(biāo)抗堿性磷酸酶(ALP)、骨鈣素(BGP)、膠原蛋白Ⅰ(CollagenⅠ)和成脂肪分化指標(biāo)脂蛋白酶(LPL)、脂肪細胞結(jié)合蛋白2(AP-2)、瘦素(Leptin)表達的影響以及對骨保護素和可溶性細胞核因子-κB受體活化因子及配基(OPG/RANKL/RANK)系統(tǒng)的影響.結(jié)果顯示Sema4D基因沉默后,絕經(jīng)后骨質(zhì)疏松患者BMMSCs中ALP、BGP、CollagenⅠ的表達水平明顯升高,LPL、AP-2、Leptin的表達水平也明顯升高;OPG的表達水平明顯升高,而RANKL和RANK的表達水平明顯下降.由上述結(jié)果可知,Sema4D基因可以抑制BMMSCs成骨分化和成脂肪分化相關(guān)基因的表達,可能通過下調(diào)OPG/RANKL/RANK系統(tǒng)相關(guān)基因的表達來實現(xiàn).
[Abstract]:To investigate the effect of Sema4D gene on the osteogenesis and adipogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) in postmenopausal patients with osteoporosis, three siRNAs, fragments of Sema4D gene were designed. The morphology of BMMSCs and the expression of Sema4D protein were identified by microscopical observation and Western blot (Western blot). Furthermore, real-time fluorescence quantitative PCR (real-time PCR) and Western blot were used to detect Sema4D gene silencing and osteogenic index (ALP), osteocalcin (BGP), collagen 鈪，

本文編號：2390085