肾小管上皮细胞转分化过程中ILK表达变化及大黄素对其干预效.docVIP

肾小管上皮细胞转分化过程中ILK表达变化及大黄素对其干预效 　 作者：陈廷芳, 陈明, 秦建华, 王明勇 【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）, Plt;0.05］, 减少Ecadherin的表达［（82.5±1.0） vs （36.0±2.8）, Plt;0.05), 促进ILK的表达［（36.1±3.1） vs （82.4±1.2）, Plt;0.05］, 促进FN的合成［（54.6±3.1） mg/L vs （124.8±3.2） mg/L, Plt;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。 【关键词】 白细胞介素 1β 大黄素 ILK 转分化 　　［Abstract］ AIM: To observe the change of ILK expression in interleukin1β(IL1β)induced tubular epithelialmyofibroblast transdifferentiation, and to investigate whether emodin inhibit IL1βinduced tubular epithelialmyofibroblast transdifferentiation 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, Plt;0.05), the expression of Ecadherin was decreased (82.5±1.0 vs 36.0±2.8, Plt;0.05), the expression of ILK was enhanced (36.1±3.1 vs 82.4±1.2, Plt;0.05), and the secretion of FN was increased (54.6±3.1 vs 124.8±3.2 mg/L, Plt;0.05). Emo