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目的:研究转IgG启动子调控GFP基因表达HepG2细胞激活过程中相关炎性因子表达的变化。方法:采用E lisa法评价葛根素和LPS后诱导IgG启动子转基因细胞分泌IL-1,βIL-8,TNF-α和MCP-1蛋白量,qPCR法评价炎性相关因子和固有免疫相关因子的mRNA转录表达量。结果:和HepG2细胞相比,IgG启动子转基因细胞不增加炎性因子分泌和基因表达量,不激活固有免疫。葛根素不增加转基因细胞炎性因子分泌和基因表达量,不激活固有免疫系统。LPS激活固有免疫系统,显著升高IL-8,TNF-α和MCP-1分泌量。结论:IgG启动子转基因HepG2细胞可以作为评价II型变态反应的特异性细胞模型,提示葛根素可以作为特异性激活IgG启动子的阳性对照品。
Objective: To study the changes of the expression of inflammatory cytokines involved in the activation of GFP gene HepG2 cells mediated by transfection of IgG promoter. Methods: The levels of IL-1, βIL-8, TNF-α and MCP-1 secreted by IgG promoter transgenic cells induced by puerarin and LPS were evaluated by ELISA. The levels of inflammatory factors and innate immunity-related factors were evaluated by qPCR mRNA transcriptional expression. Results: Compared with HepG2 cells, IgG promoter transgenic cells did not increase the secretion of inflammatory cytokines and gene expression, and did not activate innate immunity. Puerarin does not increase the secretion of inflammatory cytokines and gene expression in transgenic cells, does not activate the innate immune system. LPS activated the innate immune system, significantly increased the secretion of IL-8, TNF-α and MCP-1. Conclusion: The IgG promoter HepG2 cells can be used as a specific cell model to evaluate the type II allergy, suggesting that puerarin can be used as a specific positive control IgG promoter.