IL-18R通过Rho家族GTP酶调控中性粒细胞迁移在肝缺血再灌注损伤中的作用机制研究

Hepatic ischemia-reperfusion injury (IRI) is an important factor leading to postoperative liver dysfunction. During reperfusion injury, neutrophils mobilized from bone marrow to ischemic pretreated liver and directly lead to liver injury. In our previous studies, we found that the expression of IL-18R on the neutrophil surface of liver transplant recipients increased significantly after the gene chip screening. At the same time, the migration rate of neutrophils increased. It is speculated that the migration of neutrophils after hepatic IRI may be related to IL-18R. This paper will explore the relationship between IL-18R and neutrophil migration through flow cytometry, RT-PCR, Transwell, siRNA interference and Rho GTPases technology.Cytoskeleton is one of the key steps in the migration of neutrophils. It is not clear whether IL-18R regulate the cytoskeletal protein by the activity of Rho GTPases.To explore its mechanism,we will identification the IL-18R and the Rho GTPases signal pathway in presence of interactions by Western Blot and Co-IP. Finally, we will verified the previous results by KO mouse liver IRI model. Thus providing a new way of thinking for the treatment of patients with liver IRI.

肝缺血再灌注损伤（IRI）是导致术后肝功能障碍的重要因素，再灌注损伤时中性粒细胞从骨髓动员到缺血预处理的肝脏中，直接导致肝损伤。前期研究中，我们通过基因芯片筛查发现肝移植患者术后中性粒细胞表面IL-18R表达显著增高，且中性粒细胞迁移率增加，推测肝IRI后中性粒细胞的迁移可能与IL-18R有关。本课题将通过流式细胞术、RT-PCR、Transwell小室、siRNA干扰、慢病毒转染等技术验证IL-18R与中性粒细胞迁移的关系；中性粒细胞迁移与细胞骨架密切相关，IL-18R是否直接影响Rho GTPases活性调控细胞骨架蛋白尚不清楚。我们采用Western Blot和Co-IP 鉴定分析IL-18R与Rho GTPases相应的信号通路是否存在相互作用，从而阐明其分子机制；最后构建KO小鼠肝IRI模型在体验证之前的研究结果，从而为肝IRI患者的治疗提供新的思路。

肝缺血再灌注损伤（IRI）是导致术后肝功能障碍的重要因素，再灌注损伤时中性粒细胞从骨髓动员到缺血预处理的肝脏中，直接导致肝损伤。前期研究中，我们通过基因芯片筛查发现肝移植患者术后中性粒细胞表面IL-18R表达显著增高，且中性粒细胞迁移率增加，推测肝IRI后中性粒细胞的迁移可能与IL-18R有关。本课题通过流式细胞术、RT-PCR、Transwell小室、siRNA干扰等技术验证了 IL-18R与中性粒细胞迁移的关系；中性粒细胞迁移与细胞骨架密切相关，我们采用Western Blot和Co-IP 鉴定分析IL-18R与Rho GTPases相应的信号通路存在相互作用，从而初步阐明了其分子机制；部分结果已经发表SCI文章一篇，从而为肝IRI患者的治疗提供了新的临床思路。