血根碱在小鼠烟曲霉菌性角膜炎中的抗炎作用

ANTI-INFLAMMATORY EFFECT OF SANGUINARINE AGAINST MOUSE ASPERGILLUS FUMIGATUS KERATITIS

目的探索血根碱(SAN)在小鼠烟曲霉菌感染角膜炎模型中的抗炎作用。方法应用CCK-8实验检测不同浓度SAN(0、62.5、125.0、250.0、500.0、1000.0μg/L)对小鼠来源巨噬细胞(RAW264.7)增殖活力的影响;制备小鼠烟曲霉菌角膜炎模型,取感染第3天眼球,应用苏木精-伊红(HE)染色检测小鼠角膜组织炎症细胞浸润程度,ELISA法检测小鼠角膜组织肿瘤坏死因子α(TNF-α)及环氧化酶2(COX-2)的蛋白表达水平;采用RT-PCR和ELISA方法检测巨噬细胞中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、TNF-α及单核细胞趋化蛋白(MCP-1)的蛋白和mRNA的表达。结果500.0μg/L的SAN对RAW264.7细胞活性无影响(F=0.292,P>0.05);HE染色显示,500.0μg/L SAN组小鼠角膜组织炎症细胞浸润相较于模型组明显减少;500.0μg/L SAN可显著下调烟曲霉菌诱导的RAW264.7细胞中IL-1β、IL-6、TNF-α、MCP-1的mRNA和蛋白表达及小鼠模型中TNF-α、COX-2的蛋白表达水平,差异均有显著性(F=20.939~2690.170,P<0.01)。结论SAN通过抑制炎症细胞浸润和下调炎症递质的表达在烟曲霉菌感染中发挥抗炎作用。

Objective To explore the anti-inflammatory effect of sanguinarine(SAN)in a mouse model of Aspergillus(A.)fumigatus keratitis.Methods The effects of SAN at different concentrations(0,62.5,125.0,250.0,500.0,1000.0μg/L)on the proliferative activity of mouse-derived macrophages(RAW264.7)were determined by using Cell Counting Kit-8.A mouse model of A.fumigatus keratitis was prepared.The eyes were taken on the third day of infection.The degree of inflammatory cell infiltration in the mouse cornea was determined with hematoxylin-eosin(HE)staining.The protein expression of tumor necrosis factor-α(TNF-α)and cyclooxygenase-2(COX-2)in the cornea was measured by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expression levels of interleukin 1β(IL-1β),interleukin 6(IL-6),TNF-α,and monocyte chemotactic protein-1(MCP-1)in macrophages were measured by RT-PCR and ELISA.Results SAN at 500.0μg/L had no significant effect on the activity of RAW264.7 cells(F=0.292,P>0.05).HE staining revealed a significant decrease in inflammatory cell infiltration in the cornea in the SAN(500.0μg/L)group compared with the model group;SAN at 500.0μg/L significantly down-regulated the mRNA and protein expression of IL-1β,IL-6,TNF-α,and MCP-1 in A.fumigatus-induced RAW264.7 cells and the protein le-vels of TNF-αand COX-2 in the mouse model(F=20.939-2690.170,P<0.01).Conclusion SAN exerted anti-inflammatory effects against A.fumigatus infection via inhibiting inflammatory cell infiltration and down-regulating the expression of inflammatory mediators.