摘要
目的:研究信号蛋白ILK在IL-1β诱导的肾小管上皮细胞-肌成纤维细胞转分化(TEMT)中的表达变化以及大黄素是否是通过抑制ILK的表达而影响IL-1β诱导的肾小管上皮细胞-肌成纤维细胞转分化。方法:以体外培养的正常大鼠肾小管上皮细胞株(NRK52E)为研究对象,分为空白对照组、大黄素对照组、IL-1β诱导组、IL-1β加大黄素组。在倒置相差显微镜下观察细胞形态变化;免疫双标法检测a-平滑肌肌动蛋白(a-SMA)和E-钙黏连素(E-cadherin)的表达;免疫单染检测ILK的表达;ELISA法测定培养细胞分泌的纤维连接蛋白(FN)含量。结果:IL-1β诱导细胞转变为明显类似成纤维细胞形态,增加a-SMA的表达[(65.5±1.7)vs(140.4±3.0),P<0.05],减少E-cadherin的表达[(82.5±1.0)vs(36.0±2.8),P<0.05),促进ILK的表达[(36.1±3.1)vs(82.4±1.2),P<0.05],促进FN的合成[(54.6±3.1)mg/Lvs(124.8±3.2)mg/L,P<0.05],加入大黄素后上述变化受到明显抑制。结论:在IL-1β诱导的TEMT中ILK的表达是明显上调的,大黄素可能通过下调ILK的表达而抑制IL-1β诱导的TEMT。
AIM： To observe the change of ILK expression in intedeukin-1β （ IL-1β） -induced tubular epithelial-myo- fibroblast transdifferentiation, and to investigate whether emodin inhibit IL-1β-induced tubular epithelial-myofibroblast transdifferenUation through an intergern linked kinase-dependent mechanism. METHODS： Normal rat kidney epithelial cell line （NRK52E） was cultured and then divided into blank group, emodin control group, IL-1β-induced group and emodin-inhibited group. When the cells were cultured for 48 h, their morphological changes were observed by an inverted phase contrast microscope. The expression of a-smooth muscle actin （a-SMA） and E-cadherin were detected using a two-color immunohistochemistry staining technique, while the expression of integrin-linked kinase （ILK） was detected using a one-color immunohistochemistry staining technique. The secretion of fibronectin （FN） was analyzed by ELISA. RESULTS： NRK52E cells cultured with IL-1β became fibroblast-like in appearance. The expression of a-SMA was enhanced （65.5 ± 1.7 vs 140. 4 ±3.0, P〈 0. 05）, the expression of E-cadherin was decreased （82. 5 ± 1.0 vs 36. 0 ± 2. 8, P 〈 0. 05 ）, the expression of ILK was enhanced （36.1 ±3.1 vs82.4 ±1.2, P〈0.05）, and the secretion of FN was increased （54. 6 ± 3.1 vs 124.8 ± 3.2 mg/L, P 〈 0. 05）. Emodin markedly inhibited all of those changes induced by IL-1β. CONCLUSION： The expression of ILK is up-regulated in IL-1β-induced tubular epithelialmyofibroblast transdifferenUation. Emodin might inhibit TEMT by a down-regulation the expression of ILK.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2008年第6期574-576,583,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
四川省教育厅重点资助项目(2004049)
