甘草查尔酮A对小鼠真菌性角膜炎的抗炎作用

Anti-inflammatory effect of Licochalcone A on fungal keratitis in mice

目的 探讨甘草查尔酮A(LA)在小鼠烟曲霉菌(AF)性角膜炎中的抗炎作用。方法 采用CCK-8实验检测不同浓度的LA溶液(12、24、36、48、60μmol/L)对小鼠来源巨噬细胞(RAW264.7)增殖活力的影响;采用免疫荧光法检测AF感染后第3天小鼠角膜中性粒细胞的浸润程度;使用实时荧光定量PCR和酶联免疫吸附试验方法检测各组RAW264.7细胞中白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和血红素加氧酶(HO-1)的mRNA及蛋白表达水平。结果 60μmol/L的LA对RAW264.7细胞没有毒性(P>0.05),并显著降低感染后小鼠角膜的中性粒细胞浸润(t=5.94,P<0.01);60μmol/L的LA显著下调AF诱导的IL-6、IL-1β、TNF-α的mRNA和蛋白表达(F=15.35~507.53,P<0.001)并上调HO-1 mRNA表达(F=1 633.06,P<0.001)。结论 LA能够通过减少中性粒细胞浸润、下调炎症因子表达和增强HO-1表达,在AF性角膜炎中起到抗炎作用。

Objective To investigate the anti-inflammatory effect of Licochalcone A(LA)on Aspergillus fumigatus(AF)keratitis in mice.Methods CCK-8 assay was used to observe the effect of different concentrations of LA solution(12,24,36,48,and 60μmol/L)on the proliferative activity of murine macrophages(RAW264.7);immunofluorescence assay was used to measure the degree of neutrophil infiltration in the cornea of mice on day 3 of AF infection;quantitative real-time PCR and enzyme-linked immunosorbent assay were used to measure the mRNA and protein expression levels of interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and heme oxygenase(HO-1)in RAW264.7 cells.Results LA at the concentration of 60μmol/L had no toxicity to RAW264.7 cells(P>0.05)and significantly reduced neutrophil infiltration in the cornea after infection(t=5.94,P<0.01).LA at the concentration of 60μmol/L significantly downregulated the mRNA and protein expression levels of IL-6,IL-1β,and TNF-αinduced by AF(F=15.35-507.53,P<0.001)and upregulated the mRNA expression of HO-1(F=1633.06,P<0.001).Conclusion LA exerts an anti-inflammatory effect in AF infection by reducing neutrophil infiltration,downregulating the expression of inflammatory factors,and enhancing the expression of HO-1.