本文選題：成骨細(xì)胞 + BMSCs�。� 參考：《中國修復(fù)重建外科雜志》2016年06期

【摘要】：目的探討B(tài)MSCs的旁分泌作用對地塞米松所致成骨細(xì)胞成骨功能抑制作用的影響。方法首先通過培養(yǎng)小鼠BMSCs 24 h制備無血清條件培養(yǎng)基備用。將小鼠MC3T3-E1細(xì)胞株復(fù)蘇并傳至第3代用于實(shí)驗(yàn),分為4組:A組為對照組;B組于細(xì)胞中添加1μmol/L地塞米松;C組于細(xì)胞中按1∶1比例添加1μmol/L地塞米松和BMSCs條件培養(yǎng)基;D組僅添加BMSCs條件培養(yǎng)基。培養(yǎng)24 h后收集細(xì)胞測定ALP含量,Western blot測定細(xì)胞內(nèi)RUNX2、骨鈣素(Osteocalcin)蛋白表達(dá),實(shí)時熒光定量PCR(real-time fluorescence quantitative PCR,RT-q PCR)測定細(xì)胞內(nèi)α1-Ⅰ型膠原(collagen typeⅠ-α1,COL1A1)、RUNX2、ALP、Osteocalcin基因表達(dá);21 d后行茜素紅染色觀察鈣結(jié)節(jié)形成情況。結(jié)果培養(yǎng)24 h后,B、C、D組ALP含量均顯著低于A組,B組低于C、D組(P0.05),C、D組間差異無統(tǒng)計學(xué)意義(P0.05)。Western blot檢測示,B組RUNX2蛋白相對表達(dá)量顯著低于A、C、D組(P0.05),A、C、D組間比較差異無統(tǒng)計學(xué)意義(P0.05);B組Osteocalcin蛋白相對表達(dá)量均顯著低于A、C、D組,A、C組低于D組(P0.05),A、C組間差異無統(tǒng)計學(xué)意義(P0.05)。RT-q PCR檢測示,B、C、D組RUNX2、Osteocalcin、COL1A1、ALP基因相對表達(dá)量均顯著低于A組(P0.05);D組RUNX2、Osteocalcin、ALP基因相對表達(dá)量均顯著高于B、C組,C組顯著高于B組(P0.05);D組COL1A1基因相對表達(dá)量顯著高于B組(P0.05),B、C組間及C、D組間比較差異均無統(tǒng)計學(xué)意義(P0.05)。培養(yǎng)6 d時A組細(xì)胞死亡;21 d時B、C、D組均可見鈣結(jié)節(jié)陽性染色,且逐漸增強(qiáng)。結(jié)論小鼠BMSCs條件培養(yǎng)基能改善地塞米松對成骨細(xì)胞成骨功能的抑制效應(yīng)。
[Abstract]:Objective to investigate the effect of paracrine of BMSCs on osteogenic inhibition of osteoblasts induced by dexamethasone. Methods BMSCs were cultured for 24 h to prepare serum-free conditioned medium. The mouse MC3T3-E1 cell line was resuscitated and transferred to the third passage. Group B added 1 渭 mol / L dexamethasone to the cells of group B and group D added 1 渭 mol / L dexamethasone and BMSCs to the conditioned medium at 1:1. After 24 hours of culture, the content of ALP and the expression of osteocalcin protein were determined by Western blot. The intracellular 偽 1- 鈪，

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