中文摘要：

目的：构建两个高表达人TNFα和IL-1β细胞系,建立抗炎药物筛选细胞模型。方法：运用PCR的方法从载体pCMVSport-TNFα和p CMVSport-IL1β上扩增目的基因,以亚克隆方法将目的基因分别插入真核表达载体pcDNA3.1和pFLAG-CMV中,用单酶切、PCR扩增和基因测序的方法鉴定重组效果,然后将重组成功的质粒转入HEK293细胞系内,挑选能够稳定表达并遗传的单克隆细胞株,用蛋白免疫印迹（Western blot）法分析其表达效果。结果：三种鉴定方法均显示重组质粒构建成功。Western blot结果显示,细胞株T3、T4均能较高表达炎症因子TNF-α;细胞株I2、I3、I5均能较高表达炎症因子IL-1β。结论：成功构建了TNFα和IL-1β靶标的药物筛选细胞模型,为筛选具有抗炎作用的中药提供了一个新平台。

英文摘要：

Objective： Establish a novel cell-based model for drug screening targeting human TNFα and IL-1β. Methods： Clone the target genes of TNF-α and IL-1β from vectors p CMVSport-TNFα and p CMVSport-IL1β. The target genes were connected to p FLAG-CMV and pc DNA3.1 respectively. Single endonuclease digestion, PCR reactions and gene sequencing were used to verify whether the recombinant plasmids were constructed successfully and then they were stable transfected into HEK293 cell line. Detect the expression level of TNFα and IL-1β in monoclonal with the method of Western blot. Results： Recombinant plasmid was successfully constructed. Results of Western blot showed that cell line T3 and T4 can be highly express inflammatory cytokines TNF-α; cell line I2, I3 and I5 can be highly express inflammatory cytokines IL-1β. Conclusion： The HEK293 cell-based drug screening model targeting inflammatory cytokine TNFα or IL-1β has been established here and is available for the screening of anti-inflammatory drugs.