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纸质出版日期:2018,
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姜黄素抑制脂多糖诱导视网膜色素上皮细胞炎症反应[J]. 重庆医科大学学报, 2018,43(7):994.
Inhibitory effect of curcumin on inflammatory responses induced by LPS in human retinal pigment epithelial cells[J]. Journal of Chongqing Medical University, 2018,43(7):994.
目的:探讨姜黄素(curcumin)对脂多糖(lipopolysaccharide,LPS)诱导视网膜色素上皮细胞(retinal pigment epithelial cells
RPE)炎症反应的抑制作用。方法:体外培养人RPE细胞株ARPE-19,CCK-8测定不同浓度姜黄素(5、10、20、30、40 ?滋mol/L)或不同浓度姜黄素和脂多糖(5 ?滋g/mL)刺激后细胞的活性,以此确定姜黄素的最适作用浓度。实时荧光定量(real-time poly-merase chain reaction
RT-PCR)检测白细胞介素(interleukin,IL)-8和IL-6的mRNA表达变化,酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)测定IL-6和IL-8的蛋白表达水平。Western blot检测IκB-α、p-IκB-α蛋白表达变化。结果:CCK-8:不同浓度姜黄素对ARPE-19细胞活性无影响(P5 ?滋mol/L=0.998,P10 ?滋mol/L=1.000,P20 ?滋mol/L=0.124,P30 ?滋mol/L=0.092,P40 ?滋mol/L=0.078),预先作用不同浓度的姜黄素对细胞活性有保护作用,20 ?滋mol/L保护作用明显(P=0.041)。PCR:姜黄素+LPS组IL-8的mRNA相对表达量(4.965±3.863)比LPS组(106.331±37.314)明显降低(P=0.000),姜黄素+LPS组IL-6的mRNA相对表达量(0.992±0.374)比LPS组(2.175±0.560)明显降低(P=0.005)。ELISA:姜黄素+LPS组IL-8的蛋白表达量[(191.600±18.773) pg/mL]比LPS组[(589.025±38.880) pg/mL]明显降低(P=0.000),姜黄素+LPS组IL-6的蛋白表达量[(113.906±27.947) pg/mL]比LPS组[(243.380±38.001) pg/mL]明显降低(P=0.000)。Western blot:与LPS组相比较,姜黄素+LPS组的p-IκB-α蛋白的表达水平降低(P=0.001)。结论:姜黄素通过抑制NF-κB活性抑制了脂多糖诱导视网膜色素上皮细胞的炎症反应,保护视网膜色素上皮细胞。
Objective:To explore the inhibitory effect of curcumin on inflammatory responses induced by lipopolysaccharide (LPS) in retinal pigment epithelial(RPE) cells. Methods:ARPE-19 cells were cultured in vitro. Before being stimulated with or without LPS (5 μg/mL),ARPE-19 cells were pre-incubated with different concentrations of curcumin(5,10,20,30,40 ?滋mol/L). The optimum con-centration of curcumin was detected by CCK-8 assay. The mRNA level of IL-6 and IL-8 was measured by real-time polymerase chain reaction(RT-PCR) and their protein expression was detected by enzyme-linked immunosorbent assay(ELISA). Protein expres-sion levels of IκB-α and p-IκB-α were examined by Western blot. Results:The CCK-8 results showed that the activities of ARPE-19 cells were not affected by different concentrations of curcumin(P5 ?滋mol/L=0.998,P10 ?滋mol/L=1.000,P20 ?滋mol/L=0.124,P30 ?滋mol/L=0.092,P40 ?滋mol/L=0.078). Curcumin was indicated to be able to protect ARPE-19 cells since pre-treatment with different concentrations of curcumin following by LPS administration. This protection resulted with a maximal protective effect at 20 μmol/L(P=0.041). The RT-PCR results indicated that the relative mRNA level of IL-8 in curcumin+LPS group(4.965±3.863) was significantly lower than that in LPS group(106.331±37.314)(P=0.000) and the relative mRNA level of IL-6 in curcumin+LPS group(0.992±0.374) was significantly lower than that in LPS group(2.175±0.560)(P=0.005). The ELISA results showed that protein expression of IL-8 in curcumin+LPS group[(191.600±18.773) pg/mL] was significantly lower than that in LPS group[(589.025±38.880) pg/mL](P=0.000) and protein expression of IL-6 in curcumin+LPS group[(113.906±27.947) pg/mL] was significantly lower than that in LPS group[(243.380±38.001)pg/mL](P=0.000). Meanwhile,the Western blot results indicated that pre-treatment with curcumin inhibit-ed the phosphorylation of IκB-α(P=0.001) mediated by LPS stimulation. Conclusion:Curcumin inhibits the inflammatory responses induced by LPS via suppression NF-κB activity and protects retinal pigment epithelial cells.
姜黄素视网膜色素上皮细胞炎症反应
curcuminretinal pigment epithelial cellsinflammatory response
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