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目的研究姜黄素对脂多糖诱导的人肾小管上皮细胞炎症反应及生长抑制的保护作用,并初步探讨其机制。方法选用人肾小管上皮细胞系HK-2体外培养,根据实验要求设置阴性对照组、脂多糖处理组、姜黄素预保护组、姜黄素对照组。采用四甲基偶氮唑蓝(MTT)实验检测脂多糖和姜黄素对HK-2细胞增殖的影响;实时定量PCR检测HK-2细胞IL-8基因表达水平;酶联免疫吸附测定(ELISA)法检测白介素-8(IL-8)释放水平;Western-blot检测转录因子激活蛋白-1(AP-1)的表达水平。结果脂多糖处理可导致HK-2细胞生长抑制、IL-8表达释放增加及AP-1表达升高,姜黄素预处理则能够显著抑制IL-8和AP-1的表达。结论姜黄素对脂多糖诱导的HK-2细胞IL-8表达及释放具有抑制作用,提示姜黄素具有减轻肾小管上皮细胞炎性反应的作用,抑制细胞AP-1信号通路可能是其保护作用机制之一。
Objective To study the protective effect of curcumin on inflammatory reaction and growth inhibition induced by lipopolysaccharide in human renal tubular epithelial cells and to explore its mechanism. Methods Human renal tubular epithelial cell line HK-2 was cultured in vitro. According to the experimental requirements, a negative control group, a lipopolysaccharide treatment group, a curcumin pre-protection group and a curcumin control group were set up. The effect of lipopolysaccharide and curcumin on the proliferation of HK-2 cells was detected by MTT assay. The expression of IL-8 gene in HK-2 cells was detected by real-time quantitative polymerase chain reaction (RQ-PCR). The enzyme linked immunosorbent assay (ELISA) Method was used to detect the release of interleukin-8 (IL-8). The expression level of transcription factor activator protein-1 (AP-1) was detected by Western-blot. Results Lipopolysaccharide treatment could inhibit the growth of HK-2 cells, increase the expression of IL-8 and increase the expression of AP-1. Curcumin pretreatment could significantly inhibit the expression of IL-8 and AP-1. Conclusion Curcumin can inhibit the expression and release of IL-8 in HK-2 cells induced by lipopolysaccharide, suggesting that curcumin may reduce the inflammatory reaction of renal tubular epithelial cells and inhibit the AP-1 signaling pathway may be the protective mechanism one.