摘要
目的:观察白细胞介素1β对肾小管上皮细胞-肌成纤维细胞转分化及细胞分泌功能的影响。方法:实验于2005-06/2006-01在泸州医学院附属医院传染免疫研究室完成。以体外培养的正常大鼠肾小管上皮细胞(NRK52E)为研究对象,给予白细胞介素1β(10μg/L)刺激,12,24,48,72h后在倒置相差显微镜下观察细胞形态变化;免疫双标法检测α-平滑肌肌动蛋白和E-钙粘连素的表达;ELISA法测定培养细胞分泌的纤维连接蛋白含量。结果:白细胞介素1β刺激48h后肾小管上皮细胞转变为明显类似成纤维细胞形态;刺激12h即可见α-平滑肌肌动蛋白的表达由64.80±2.19增加到83.23±2.71(P<0.05),E-钙粘连素的表达由81.77±1.27减少到64.50±1.31(P<0.05),随刺激时间延长上述变化越明显,72hα-平滑肌肌动蛋白表达增加到170.53±3.94;E-钙粘连素表达减少到26.20±2.32。细胞培养上清液中纤维连接蛋白含量随时间延长增加(P<0.05),白细胞介素1β刺激72h时纤维连接蛋白增加1倍。结论:白细胞介素1β可诱导肾小管上皮细胞-肌成纤维细胞转分化,增加纤维连接蛋白的分泌。
AIM： To observe the influenCe of intedeukin （IL）-1β on tubular eplthelial-myofibroblast transdifferentiation （TEMT） and the secretion of fibronectin （FN） In cells. METHODS： The experiment was carried out in the Staff Room of Infection Immunity, Affiliated Hospital of Luzhou Medical College from June 2005 to January 2006. The normal rat kidney epithelial cell line （NRK52E） was cultured in vitro and then co-incubated with IL-1β （10 μg/L）, When the cells were cultured for 12, 24, 48, 72 hours, their morphological changes were observed through inverted phase contrast microscope; The expressions of e-smooth muscle actin （e-SMA） and E-cadherin were detected by a two-color immunohistochemistry staining technique; Supernatant liquid of every well was analyzed for secretion of FN by ELISA. RESULTS： NRK52E cells cultured with IL-1β for 48 hours became fibroblast-like in appearance; when the cells were cultured with IL-1β for 12 and 72 hours, the expression of α-SMA were enhanced （64.80±2.19, 83.23±2.71, 170.53±3.94, P 〈 0.05）, the expression of E-cadherin was decreased （81.77±1.27, 64.50±1.31, 26.20±2.32, P 〈 0.05）. These changes were time-dependent. The secretion of FN was increased with the time prolonged （P 〈 0.05） and doubled at 72 hours of culture. CONCLUSION. IL-1 β can induce TEMT and enhance the secretion of FN in cultured NRK52E cells.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第23期4470-4472,共3页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
四川省教育厅重点资助项目(2004049)~~
