摘要
目的:探讨TGF-β1对骨髓间充质干细胞(MSCs)在无氧无血清条件下凋亡的影响。方法:取大鼠骨髓,体外分离,扩增培养MSCs;TGF-β1腺病毒或对照GFP腺病毒转染MSCs。细胞分为TGF-β1-MSCs组、GFP-MSCs组,MSCs组。RT-PCR检测TGF-β1基因的表达;无氧无血清条件下分别诱导各组细胞4,8,12h,流式细胞仪检测凋亡变化;RT-PCR检测bcl-2 mRNA的表达。结果:TGF-β1转染的MSCs表达TGF-β1基因水平与GFP-MSCs组,MSCs组相比均增加(P<0.05);细胞在无氧无血清条件下诱导4,8,12h,后TGF-β1-MSCs组的凋亡率与GFP-MSCs组,MSCs组的凋亡率相比降低(P<0.05),且随时间的推移呈升高的趋势(P<0.05);bcl-2 mRNA在无氧无血清条件下诱导4,8,12h后的表达水平TGF-β1-MSCs与GFP-MSCs、MSCs相比均增加(P<0.05),且随时间的推移均呈降低趋势(P<0.05)。结论:TGF-β1修饰的MSCs可稳定高效表达TGF-β1基因;TGF-β1-MSCs通过上调bcl-2 mRNA水平抑制无氧无血清诱导的凋亡。
Objective： To investigate the effects of TGF-β1 on the levels of apoptosis of MSCs treated with hypoxia plus serum deprivation. Methods： MSCs were cultured in vitro, and transfected by Adv-TGF-β1 or Adv-GFP. RT-PCR was used to determine the expression of TGF-β1. FACS was used to determined the apoptosis of MSCs under the condition of hypoxia plus serum deprivation at 4, 8 and 12 h, respectively, and RT-PCR was used to determine the expression of bcl-2 mR- NA. Results： TGF-β1-MSCs exhibited increased TGF-β1 expression. The levels of apoptosis at 4, 8 and 12 h after treatment with hypoxia plus serum deprivation were all significantly decreased in TGF-β1-MSCs than those in GFP-MSCs and MSCs, and increased with time. The levels of bcl-2 mRNA expression at 4, 8 and 12 h were all significantly decreased in TGF-β1-MSCs than those in GFP-MSCs and MSCs, and decreased with time. Conclusion： TGF-β1-MSCs exhibited increased TGF-β1 expression, and decreased the levels of apoptosis of MSCs treated with hypoxia plus serum deprivation by upregulating bcl-2 mRNA.
出处
《武汉大学学报（医学版）》
CAS
2007年第6期729-732,共4页
Medical Journal of Wuhan University
