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目的研究新的非甾体类抗炎药利克飞龙(Licofelone)对大鼠肾小球系膜细胞IL-18诱导的趋化因子Fractalkine表达的影响。方法体外培养大鼠肾小球系膜细胞。设立IL-18刺激组(加入10μg/LIL-18刺激肾小球系膜细胞24h)、Licofelone干预组(先用10、50、100μmol/L的Licofelone处理肾小球系膜细胞30min,再加入10μg/LIL-18刺激肾小球系膜细胞24h)、正常对照组(无IL-18刺激和Licofelone干预,加等量的9g/L盐水作为对照)。反转录-多聚酶链反应(RT-PCR)测定各组FractalkinemRNA表达量。ELISA法测定各组细胞培养上清液中Fractalkine蛋白量。结果正常对照组FractalkinemRNA的相对表达量为179.0±21.0,IL-18刺激24h后,肾小球系膜细胞中趋化因子FractalkinemRNA表达上调为1220.1±185.7,与正常对照组比较有显著性差异(t=9.646P<0.01)。10、50、100μmol/L的Licofelone呈剂量依赖性拮抗IL-18诱导的FractalkinemRNA表达上调,FractalkinemRNA的相对表达量依次为899.7±85.3、369.6±36.7、236.6±35.6,与IL-18刺激组比较有显著性差异(t=2.798、7.780、9.006Pa<0.05)。IL-18刺激24h,肾小球系膜细胞中趋化因子Fractalkine蛋白表达量为(2097.4±293.8)ng/L,显著高于正常对照组(380.4±31.1)ng/L(t=10.068P<0.01)。10、50、100μmol/L的Licofelone呈剂量依赖性拮抗IL-18诱导的Fractalkine蛋白表达上调,细胞培养上清中Fractalkine蛋白量依次为(1257.9±245.6)、(908.2±155.3)、(650.7±126.6)ng/L,与IL-18刺激组比较有显著性差异(t=3.798、6.199、7.834Pa<0.05)。结论Licofelone拮抗肾小球系膜细胞中IL-18诱导的趋化因子Fractalkine基因表达。Licofe-lone在治疗免疫性肾损伤方面有一定的应用前景。
Objective To investigate the effect of Licofelone, a new non-steroidal anti-inflammatory drug, on the expression of Fractalkine induced by IL-18 in rat mesangial cells. Methods Rat glomerular mesangial cells were cultured in vitro. The IL-18 stimulation group (stimulating mesangial cells with 10μg / L IL-18 for 24h) and Licofelone intervention group (mesangial cells were treated with Licofelone at 10, 50 and 100μmol / L for 30min, / LIL-18 stimulation of mesangial cells 24h), normal control group (without IL-18 stimulation and Licofelone intervention, plus an equal amount of 9g / L saline as a control). Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of Fractalkine mRNA in each group. The amount of Fractalkine protein in each cell culture supernatant was determined by ELISA. Results The relative expression of Fractalkine mRNA in normal control group was 179.0 ± 21.0. The mRNA expression of Fractalkine in mesangial cells was up-regulated to 1220.1 ± 185.7 after stimulation with IL-18 for 24 hours, which was significantly different from the normal control group (t = 9.646P <0.01). At doses of 10, 50 and 100μmol / L, Licofelone antagonized IL-18-induced up-regulation of Fractalkine mRNA in a dose-dependent manner. The relative expression of Fractalkine mRNA was 899.7 ± 85.3, 369.6 ± 36.7 and 236.6 ± 35.6, Significant difference (t = 2.798,7.780,9.006Pa <0.05). The expression of Fractalkine protein in mesangial cells was (2097.4 ± 293.8) ng / L, which was significantly higher than that of normal control group (380.4 ± 31.1) ng / L (t = 10.068P < 0.01). At doses of 10, 50 and 100μmol / L Licofelone antagonized the up-regulation of Fractalkine induced by IL-18 in a dose-dependent manner. The levels of Fractalkine in the cell culture medium were (1257.9 ± 245.6), (908.2 ± 155.3), ) ng / L, there was a significant difference compared with IL-18 stimulation group (t = 3.798,6.199,7.834Pa <0.05). Conclusion Licofelone antagonizes IL-18-induced chemokine Fractalkine gene expression in mesangial cells. Licofe-lone in the treatment of immune kidney injury has a certain application prospects.