基于斑马鱼模型的纯绿青霉醇抗炎活性研究

Anti-inflammatory activity of viridicatol based on zebrafish model

目的利用斑马鱼急性炎症模型,对纯绿青霉醇(viridicatol)抗炎作用及其机制进行初步研究。方法采用硫酸铜(CuSO4)构建斑马鱼急性炎症模型,评价纯绿青霉醇的抗炎活性。将炎症状态下的斑马鱼(3 dpf)暴露在不同浓度的纯绿青霉醇(30、60和90μg/mL)中2 h,布洛芬(ibuprofen,IBF)作为阳性对照,观察斑马鱼神经侧线炎症细胞迁移和聚集的数量变化情况并计算荧光强度;利用RT-PCR技术测定各组斑马鱼炎症相关基因的转录水平。结果与炎症模型组相比,布洛芬组和纯绿青霉醇组斑马鱼炎症细胞迁移和聚集的数量以及荧光强度均显著降低,同时30、60、90μg/mL剂量下的纯绿青霉醇能显著提高pparγ的mRNA表达水平,抑制炎症相关因子iκbαa、ap-1、nf-κb、il-1b、il8、ptges和myd88的mRNA表达。结论本研究通过建立斑马鱼急性炎症模型探究了纯绿青霉醇的抗炎作用,首次发现其机制可能是纯绿青霉醇激活pparγ表达,抑制nf-κb和ap-1转录活性,降低炎症因子表达,从而缓解炎症,为纯绿青霉醇的应用开发提供了新思路。

Objective The anti-inflammatory effect and mechanism of viridicatol were studied by using zebrafish acute inflammation model.Methods The acute inflammation model of zebrafish was constructed with CuSO4 to evaluate the anti-inflammatory activity of viridicatol.zebrafish(3 dpf)in inflammatory state were exposed to different concentrations of viridicatol(30,60,or 90μg/mL)for two hours,with ibuprofen as a positive control.The migration and aggregation of inflammatory cells in zebrafish nerve lateral line were observed,and the fluorescence intensity was calculated.The transcription levels of inflammation related genes in zebrafish were measured by RTPCR.Results Compared with the inflammation model group,the number and fluorescence intensity of inflammatory cell migration and aggregation in zebrafish in ibuprofen group and viridicatol group were significantly reduced.At the same time,viridicatol at the dose of 30,60,or 90μg/mL could significantly increase the mRNA level of pparγand inhibit the mRNA expression of inflammation related factors,such as iκbαa、ap-1、nf-κb、il-1b、il8、ptges and myd88.Conclusion This study explored the anti-inflammatory effect of viridicatol by establishing an acute inflammation model of zebrafish.It was found for the first time that the mechanism may be viridicatol activating the expression of pparγ,inhibiting the transcription activities of nf-κb and ap-1,and reducing the expression of inflammatory factors,so as to alleviate inflammation,which provides a new idea for the application and development of viridicatol.