重组旋毛虫53-kDa蛋白对巨噬细胞表型影响的研究

Studies show helminth or helminth antigen have protection on inflammatory disease, such as inflammatory bowel disease, multiple sclerosis, even sepsis. However, helminth worm infections cannot be accepted by patients. The scanty source of antigens obtained from worms limits its practical application. So, we get recombinant Trichinella spiralis (TS) 53-kDa protein (rTsP53) by cloning, expression, purification and identification and have shown the protective effect of rTsP53 on experimental colitis in mice. In order to further study, we will detect if rTsP53 act on macrophage and how to affect macrophage phenotype and cytokines. We also detect the effect of rTsP53 on LPS induction macrophage in order to explain antiinflammation in vitro. Next, we detect STAT6 expression during rTsP53 inducing macrophage and macrophage change by rTsP53 after RNAi-STAT6. We also detect PI3K expression during rTsP53 inducing macrophage and macrophage change by rTsP53 after LY294002(PI3K inhibitor). We sudy the mechanism in depth and wish to provide new anti-inflmmation strategy.

研究表明蠕虫或蠕虫抗原对炎症性疾病如：炎症性肠病、多发性硬化,甚至脓毒症有保护作用，但用蠕虫直接作为治疗并不能被接受或蠕虫抗原资源的缺乏，给可能的临床应用带来困难。本课题组前期利用基因工程重组合成、纯化和鉴定后获得重组旋毛虫53-kDa蛋白（rTsP53）并揭示其对实验性结肠炎有保护作用。 为进一步深入探讨，我们将在细胞水平观察rTsP53能否作用于小鼠巨噬细胞，以及对巨噬细胞表型和细胞因子表达功能的影响；观察rTsP53对LPS诱导巨噬细胞因子表达的影响，从而在体外实验来阐述其抗炎作用。观察磷酸化STAT6在rTsP53作用于巨噬细胞中的表达，以及RNAi干扰STAT6后rTsP53作用巨噬细胞的变化；观察PI3K在rTsP53作用于巨噬细胞中的表达，LY294002(PI3K抑制剂)对rTsP53作用巨噬细胞的影响；进一步进行机制探讨，以期能为抗炎治疗提供新的策略。