摘要
目的探讨碱性成纤维细胞生长因子(bFGF)对胰腺星状细胞(PSCs)增殖作用的影响。方法采用组织块分离法培养PSCs,通过免疫荧光检测结蛋白、神经胶质原纤维酸性蛋白、α-平滑肌肌动蛋白(α-SMA)的表达鉴别。取培养的第2-3代PSCs,用不同浓度bFGF(5、10、20、25ng/ml)处理72h,CCK8法检测PSCs增殖情况,荧光定量PCR检测Ⅰ型胶原蛋白基因mRNA表达,Western blot方法检测α-SMA表达;其结果与对照组比较。结果 (1)不同浓度bFGF组的细胞增殖率明显高于对照组[(145.17±6.67)%、(152.96±7.50)%、(148.64±8.74)%和(144.83±10.06)%vs.100%](P<0.05)。(2)与对照组比较,bFGF促进Ⅰ型胶原蛋白mRNA表达显著增加(P<0.05)。(3)与对照组比较,bFGF刺激PSCs后引起α-SMA表达显著增加(P<0.05)。结论 bFGF可促进PSCs增殖活化,在一定程度上可促进胰腺纤维化。
Objective To investigate the effect of basic fibroblast growth factor（bFGF） on the proliferation of the pancreatic stellate cells （PSCs）. Methods PSCs were isolated and cultured with tissue culture method and were identified＇ by testing Desmin, GFAP and α-SMA with immunofluorescence. The isolated PSCs of the 2nd or 3rd generation were processed with different concentrations of hFGF 5,10,20,and 25 ng/ml for 72 hours, and then the cell proliferation was tested by CCK8 method. The mRNA expression of collagen I gene was detected by FQ-PCR and the protein expression of 0cSMA was detected by Western blot. Results The cell proliferations of different bFGF concentration groups were （145.17±6.67）%,（152.96±7.50）%, （148.64±8.74）% and （144.83±10. 06）%, respectively, which were significantly higher than 100% of control group（P〈0. 05）. The mRNA expression of collagen I gene was higher in bFGF groups than that in control group （P〈0. 05）. So did that of α-SMA stimulated by bFGF（P〈0. 05）. Conclusion bFGF can promote the activation and proliferation of the pancreatic stellate cells and can accelerate, to a certain extent, the development of pancreatic fibrosis.
出处
《江苏医药》
CAS
北大核心
2014年第11期1256-1259,F0003,共5页
Jiangsu Medical Journal
基金
江苏省自然科学基金(BK2013247)
