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目的:探讨脂氧素A4(LXA4)对结缔组织生长因子(CTGF)诱导的肾小管上皮细胞HK2转分化的影响。方法:在体外培养的人肾小管上皮细胞(HK2细胞)中,加入CTGF和LXA4刺激48h后,用倒置显微镜观察细胞形态学变化,应用RT-PCR、Westernblot方法测定上皮细胞E钙黏蛋白和α-平滑肌肌动蛋白(α-SMA)mRNA与蛋白的表达。结果:CTGF刺激使HK2细胞形态由椭圆形转为梭形,下调E钙黏蛋白mRNA与蛋白表达,上调α-SMA的mRNA与蛋白表达。LXA4能够抑制CTGF所致的上述变化,E钙黏蛋白mRNA与蛋白的相对表达值分别为0.270±0.082和0.827±0.177,分别高于CTGF刺激组的0.067±0.015(P<0.05)和0.257±0.221(P<0.05);α-SMA的mRNA与蛋白表达的相对表达值分别为0.070±0.046和0.023±0.023,分别低于CTGF刺激组的0.653±0.256(P<0.05)和0.167±0.050(P<0.05)。结论:外源性CTGF能导致HK2细胞转分化为肌成纤维细胞,LXA4可抑制CTGF对HK2细胞的转分化,这为治疗肾间质纤维化提供了一条新途径。
Objective: To investigate the effect of lipoxin A4 (LXA4) on the transdifferentiation of renal tubular epithelial cells HK2 induced by connective tissue growth factor (CTGF). Methods: Human renal tubular epithelial cells (HK2 cells) cultured in vitro were stimulated with CTGF and LXA4 for 48 hours. The morphological changes of the cells were observed by inverted microscope. The expressions of E-cadherin and α in epithelial cells were detected by RT-PCR and Western blotting Expression of smooth muscle actin (α-SMA) mRNA and protein. Results: CTGF stimulation changed the morphology of HK2 cells from oval to spindle shape, down-regulated E-cadherin mRNA and protein expression, and up-regulated the mRNA and protein expression of α-SMA. LXA4 could inhibit the above changes caused by CTGF, and the relative expression values of E-cadherin mRNA and protein were 0.270 ± 0.082 and 0.827 ± 0.177, respectively, which were higher than that of CTGF stimulation group (0.067 ± 0.015 and 0.257 ± 0.221 (P <0.05). The relative expressions of α-SMA mRNA and protein were 0.070 ± 0.046 and 0.023 ± 0.023, respectively, which were lower than those of CTGF stimulation group (0.653 ± 0.256 and 0.167 ± 0.050, P < 0.05). Conclusion: Exogenous CTGF can induce HK2 cells to transdifferentiate into myofibroblasts, and LXA4 can inhibit the transdifferentiation of HK2 cells induced by CTGF, which provides a new way to treat renal interstitial fibrosis.