腺苷信号通路在肾缺血再灌注损伤后肾纤维化的作用研究

Ischemia reperfusion injury is common in clinical surgery,particularly is inevitable process in kidney transplantaion. Subsequent chronic renal fibrosis is the main reason for the kidney damage and renal allograft loss. Adenosine is a very important signal molecule produced in the process of metabolism of kidney underwent ischemia. long-term accumulation of adenosine in kidney tissues will damage the kidney with chronic fibrosis. In mice kidney ischemia-reperfusion injury model, research for chronic kidney adenosine signal pathway in the process of kidney fibroblasts proliferation, tubular epithelial -messenchyal transition, the accumulation of extracellular matrix, and related kidney dysfunction, will verify the main adenosine receptor subtype activated in the process of ischemia-reperfusion injury.Application A2B blockers and in the A2B receptor gene knock out mice model, demonstrate the important role of A2B receptor pathway in kidney fibrosis originated from kidney ishchemia reperfusion injury and screening the main fibrotic mediators.It will help to specify the metabolic mechanism for chronic kidney fibrosis. Hope to find novel therapy and prevention methods for chronic kdiney fibrosis related to kidney ischemia reperfusion injury. In view of the renal allograft inevitable ischemia-reperfusion process, puts forward the preventive treatment of chronic kidney fibrosis post-transplantation and save the kidney function,expected to enlarge donor source, relieve lack of kdiney donation, and also can prolong the graft survival time.

肾缺血再灌注损伤在外科临床非常普遍，并且是肾移植不可避免的过程，继发的肾纤维化是肾功能损害及移植肾失功的主要原因。缺血缺氧条件下产生的腺苷是代谢过程中重要的信号分子，腺苷的长期积聚会对肾组织产生致纤维化损伤。在小鼠肾缺血再灌注损伤模型上研究腺苷信号通路对肾纤维化的过程中肾成纤维细胞、肾小球系膜细胞、肾小管上皮细胞-间质细胞转化、细胞外间质沉积及其伴随的肾功能的变化的影响，明确缺血再灌注损伤过程中激活的主要腺苷受体亚型；应用受体阻断剂及受体基因敲除小鼠模型进一步证明A2B受体途径在慢性肾纤维化中的重要作用并筛选主要致纤维化因子，有助于从代谢途径明确慢性肾病肾纤维化的确切机制，并为慢性肾病肾纤维化提供新的预防治疗方法，针对移植肾不可避免的缺血再灌注过程，提出预防治疗移植肾慢性纤维化失功的新的途径和方法，有望可扩大肾源，缓解肾源缺乏难题，而且可延长移植肾生存时间。

肾缺血再灌注损伤在外科临床非常普遍，并且是肾移植不可避免的过程，继发的肾纤维化是肾功能损害及移植肾失功的主要原因。缺血缺氧条件下产生的腺苷是代谢过程中重要的信号分子，腺苷的长期积聚会对肾组织产生致纤维化损伤。本项目明确了在腺苷受体A2BR亚型是小鼠肾脏、原代培养小鼠肾成纤维细胞以及人肾小管上皮细胞系中的腺苷受体在正常情况及缺血再灌注损伤后的主要亚型，且再灌注损伤后它们的A2BR上调。通过MTT实验证实A2BR受体能够促进小鼠纤维细胞增殖活化，通过qRT-PCR技术发现小鼠纤维细胞缺氧损伤后促纤维化细胞因子的生成增多。通过A2BR激动剂和抑制剂干预小鼠成纤维细胞缺血再灌注损伤模型、人肾小管上皮细胞纤维化模型，敲减人肾小管上皮细胞A2BR受体，从细胞水平证实A2BR能使促纤维化因子的产生增加，并揭示A2BR通过调控TGF-β1/samd3/p-samd3信号通路参与肾纤维化的发生与发展。通过A2BR激动剂和抑制剂干预小鼠缺血再灌注损伤模型进一步揭示A2BR能促进肾缺血再灌注损伤后肾纤维化，并在器官水平验证了A2BR通过调控TGF-β1/samd3/p-samd3信号通路参与肾纤维化的发生与发展。以上发现有助于明确A2BR促进慢性肾病肾纤维化的确切机制，并为慢性肾病肾纤维化提供新的预防治疗方法。