摘要
目的研究信号蛋白Smad3在TGF-β1促大鼠骨髓间充质干细胞分化成平滑肌样细胞中的作用。方法运用免疫细胞化学、RT—PCR证实BMSC中存在TGF-β1／smad3信号传导通路的基础上，运用反义Smad3进行干预。实验分三组：反义Smad3组（反义Smad3腺病毒转染大鼠BMSC），空载对照组（空载腺病毒转染大鼠BMSC）及未转染组；一组均予10ng／mL浓度的TGF-β1诱导1W。通过免疫细胞化学和实时定量PCR比较反义Smad3干预后各组SMut=actin的表达变化。结果经免疫细胞化学法、RT-PCR检测证明BMSC中存在TGF-β1／smad3表达。运用针对靶基因Smad3的反义技术干预后，经10ng／mLTGF-β1诱导，相比未转染组及空载组，反义smad3腺病毒转染组SMα-actin阳性细胞表达数明显减少。实时定量PCR也显示反义smad3组SMα-actinmRNA的相对含量明显低于未转染组（O．460±0．259VS2．932±0．375，P〈0．01），而空载组与未转染组相比无明显统计学差异。结论TGF-β1促BMSC成平滑肌样细胞分化的过程中受Smad3选择性调节，Smad3可能是TGF-β1促BMSC分化成平滑肌样细胞的细胞内信号途径。
Objective To observe the role of Smad3 signal transduction in TGF-β1 action. Methods TGF-β1/smad3 expression was confirmed in bone marrow mesenchymal stem cell （BMSC） by RT-PCR and immunocytochemistry. BMSC was divided into three groups： non-transfected group, experimental group （BMSC transfected with antisense Smad3 adenovirus vector） and blank group （BMSC transfected with empty adenovirus vector）. One week after 10ng/ml TGF-β1 induction, SM -actin and the rate of differentiation were measured by immunocytochemistry and real-time quantitative PCR identification. Results After 7 days induction by TGF-β1 （10ng/ml）, SMα-actin-positive cells were significantly reduced, and the SM ct-actin mRNA expression was significantly lower than the relative content of non-transfected group （0.460 0.259 vs 2.932 0.375, P 〈0.01）, while the blank group and non-transfected group had no significant statistical difference. Conclusion The differentiation of BMSC into smooth muscle-like cell could be partly inhibited by antisense Smad3 via block- ing the TGF-β1/Smad3 pathway.
出处
《老年医学与保健》
CAS
2011年第1期25-29,共5页
Geriatrics & Health Care
基金
上海市教委课题（06BZ016）
关键词
骨髓间充质干细胞
TGF-Β1
平滑肌样细胞
反义Smad3
Bone marrow mesenchymal stem cell
Transforming growth factor β1
Smooth muscle-like cell
Antisense Smad3
