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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。
OBJECTIVE: To investigate the expression of signal protein ILK in IL-1β-induced renal tubular epithelial-myofibroblast transdifferentiation (TEMT) and whether emodin affects IL-1β-induced tubular epithelial cells by inhibiting ILK expression. Cell-myofibroblast transdifferentiation. METHODS: Normal rat renal tubular epithelial cell lines (NRK52E) cultured in vitro were used as study subjects and divided into blank control group, emodin control group, IL-1β induction group, and IL-1β flavonoid group. The morphology of the cells was observed under an inverted phase contrast microscope. The expression of a-SMA and E-cadherin was detected by double immunosorbent assay. The expression of ILK was detected by immunostaining. The fibronectin (FN) content secreted by cultured cells was measured. RESULTS: IL-1β induced cell transformation into a fibroblast-like morphology, increased a-SMA expression [(65.5±1.7) vs (140.4±3.0), P<0.05], decreased E-cadherin expression [(82.5± 1.0) vs (36.0±2.8), P<0.05), promoted ILK expression [(36.1±3.1) vs (82.4±1.2), P<0.05], promoted FN synthesis [(54.6±3.1) mg/L vs. 124.8±3.2) mg/L, P<0.05]. The above changes were significantly inhibited after adding emodin. CONCLUSION: ILK expression is significantly up-regulated in IL-1β-induced TEMT. Emodin may inhibit IL-1β-induced TEMT by down-regulating ILK expression.