中南大学学报（医学版）
中南大學學報（醫學版）
중남대학학보（의학판）
JOURNAL OF CENTRAL SOUTH UNIVERSITY (MEDICAL SCIENCES)
2010年
2期
159-164
,共6页
刘小贤%张浩%孙剑%余学清%廖琴
劉小賢%張浩%孫劍%餘學清%廖琴
류소현%장호%손검%여학청%료금
腹膜间皮细胞%转分化%转化生长因子-β%RhoA
腹膜間皮細胞%轉分化%轉化生長因子-β%RhoA
복막간피세포%전분화%전화생장인자-β%RhoA
peritoneal mesothelial cell%epithelial-mesenchymal transition%transforming growth factor β%RhoA
目的:研究转化生长因子(transforming growth factor β1,TGF-β1)对大鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMCs)转分化的影响,并探讨其可能的机制.方法:体外培养SD大鼠原代腹膜间皮细胞,静止24 h后,随机分为正常对照组和TGF-β1(10 μg/L)刺激组.用TGF-β1(10 μg/L)刺激RPMCs不同时间,分别用RT-PCR方法和Western免疫印迹法检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、E-钙黏素、Ⅰ型胶原的mRNA和蛋白的表达.用Western免疫印迹法检测总RhoA的蛋白表达水平,活化的RhoA由膜蛋白提取试剂盒提取,并用Western免疫印迹法评估.结果:TGF-β1刺激RPMCs能导致E-钙黏素mRNA和蛋白表达下调;α-SMA,Ⅰ型胶原mRNA和蛋白表达上调,同时RhoA蛋白表达上调,呈时间依赖性.结论:TGF-β1诱导了大鼠腹膜间皮细胞转分化,其机制可能通过RhoA介导的信号通路.
目的:研究轉化生長因子(transforming growth factor β1,TGF-β1)對大鼠腹膜間皮細胞(rat peritoneal mesothelial cells,RPMCs)轉分化的影響,併探討其可能的機製.方法:體外培養SD大鼠原代腹膜間皮細胞,靜止24 h後,隨機分為正常對照組和TGF-β1(10 μg/L)刺激組.用TGF-β1(10 μg/L)刺激RPMCs不同時間,分彆用RT-PCR方法和Western免疫印跡法檢測α-平滑肌肌動蛋白(α-smooth muscle actin,α-SMA)、E-鈣黏素、Ⅰ型膠原的mRNA和蛋白的錶達.用Western免疫印跡法檢測總RhoA的蛋白錶達水平,活化的RhoA由膜蛋白提取試劑盒提取,併用Western免疫印跡法評估.結果:TGF-β1刺激RPMCs能導緻E-鈣黏素mRNA和蛋白錶達下調;α-SMA,Ⅰ型膠原mRNA和蛋白錶達上調,同時RhoA蛋白錶達上調,呈時間依賴性.結論:TGF-β1誘導瞭大鼠腹膜間皮細胞轉分化,其機製可能通過RhoA介導的信號通路.
목적:연구전화생장인자(transforming growth factor β1,TGF-β1)대대서복막간피세포(rat peritoneal mesothelial cells,RPMCs)전분화적영향,병탐토기가능적궤제.방법:체외배양SD대서원대복막간피세포,정지24 h후,수궤분위정상대조조화TGF-β1(10 μg/L)자격조.용TGF-β1(10 μg/L)자격RPMCs불동시간,분별용RT-PCR방법화Western면역인적법검측α-평활기기동단백(α-smooth muscle actin,α-SMA)、E-개점소、Ⅰ형효원적mRNA화단백적표체.용Western면역인적법검측총RhoA적단백표체수평,활화적RhoA유막단백제취시제합제취,병용Western면역인적법평고.결과:TGF-β1자격RPMCs능도치E-개점소mRNA화단백표체하조;α-SMA,Ⅰ형효원mRNA화단백표체상조,동시RhoA단백표체상조,정시간의뢰성.결론:TGF-β1유도료대서복막간피세포전분화,기궤제가능통과RhoA개도적신호통로.
Objective To explore the effect of transforming growth factor β1 (TGF-β1) on epithelial-mesenchymal transition in rat peritoneal mesothelial cells(RPMCs) and its mechanism.Methods Primary peritoneal mesothelial cells of SP rats were cultured in vitro. After synchronization for 24 h, RPMCs were randomly divided into 2 groups: Group A (control), Group B (TGF-β1, 10 μg/L). RPMCs were stimulated by 10 μg/L TGF-β1 for different time. The mRNA and protein expression levels of E-cadherin, α-smooth muscle actin (α-SMA) and collagenⅠwere measured by RT-PCR and Western blot, respectively. The protein expression level of total RhoA was measured by Western blot. Active RhoA was extracted by Plasma Membrane Protein Extraction Kit, and assessed by Western blot. Results TGF-β1 down-regulated mRNA and protein expression of E-cadherin in RPMCs, and upregulated mRNA and protein expression of α-SMA and CollagenⅠ. TGF-β1 stimulation elicited a robust increase in RhoA activity in a time-dependent manner. RhoA activity peaked at 1 h.Conclusion RPMCs can be transdifferentiated into myofibroblast under the effect of TGF-(β1,)and the mechanism may be related to the activation of RhoA associated signal pathway.
