转化生长因子-β_2诱导大鼠胰腺星形细胞活化及上皮间质转化的研究  

Study on transforming growth factor-β_2-induced(TGF-β_2-induced) activation and epithelial-mesenchymal transition in rat pancreatic stellate cells

作　　者：张尤历[1] 李萍[1] 王国英[1] 赵义[1] 王国乙 蔡菁[1] 郭柔玉[1] 刘鑫[1] 徐岷[1] 

机构地区：[1]江苏大学附属医院消化科,江苏镇江212001

出　　处：《中国现代医学杂志》2015年第7期7-11,共5页China Journal of Modern Medicine

基　　金：江苏省自然科学基金(No:BK20131247);镇江市"169四期工程"科研项目(No:镇人才办[2013])

摘　　要：目的探讨转化生长因子-β2(TGF-β2)诱导大鼠胰腺星形细胞(PSCs)活化及上皮间质转化(EMT)的作用。方法组织块法分离、培养及鉴定大鼠PSCs后,10 ng/ml TGF-β2刺激PSCs 72 h,光镜下观察细胞形态学改变;免疫荧光染色及Western blot法检测α-SMA、Col-Ⅰ的蛋白表达;q RT-PCR及Western blot法分别检测E-caderin、Vimentin基因及蛋白的表达;荧光显微镜观察E-caderin和Vimentin在细胞内的表达情况。结果免疫荧光染色显示新分离PSCs神经胶质原纤维酸性蛋白(GFAP)、结蛋白(desmin)表达阳性,α-SMA表达阴性,符合其生物学特性。TGF-β2处理PSCs后,光镜下观察细胞体积及胞核变大,伪足发达,脂滴消失;免疫荧光染色及Western blot结果显示α-SMA和Col-Ⅰ表达量显著增加,呈典型活化状态;q RT-PCR、Western blot及免疫荧光染色结果显示Vimentin的表达明显高于空白对照组,而E-caderin的表达明显降低(P<0.05)。结论 TGF-β2可诱导PSCs活化及上皮间质转化。【Objective】To observe the effect of TGF-β2on activation and epithelial-mesenchymal transition in rat pancreatic stellate cells. 【Methods】Rat pancreatic stellate cells were isolated and cultured by tissue method,and then stimulated by TGF-β2of 10 ng/m L for 72 hours after they were qualified. The morphology of the cells was observed under inverted phase contrast microscope, and the expression of α-SMA and Col-Ⅰwas measured by immunofluorescence staining and western blot assays. The expression of E-caderin and Vimentin was tested by q RTPCR, western blot and immunofluorescence staining methods.【Results】Immunofluorescence staining showed GFAP and desmin were positively expressed in fresh PSCs, but α-SMA was negative, this was consistently with theirs biological properties of PSCs. After PSCs were treated with TGF-β2, cells volume and nucleus were larger, pseudopodia developed and fat droplets disappeared under inverted phase contrast microscope. Immunofluorescence staining and western blot results demonstrated the expression of α-SMA and Col- Ⅰ was significantly increased, therefore PSCs presented a typical activation state. q RT-PCR, western blot and immunofluorescence staining results showed that the expression of Vimentin was significantly higher than the control group, while E-cadherin was obviously lower than the control group（P〈0.05）.【Conclusions】TGF-β2could induce activation and epithelial-mesenchymal transition of PSCs.

关 键 词：转化生长因子-Β2 胰腺星形细胞 活化 上皮间质转化 

分 类 号：R576[医药卫生—消化系统] R-332[医药卫生—内科学]