抑制TRAF6缓解高糖诱导的炎症反应和糖尿病肾病的作用机制研究

Chronic inflammation is one of the important pathological processes of diabetic nephropathy (DN). It will provide new strategy for treating and preventing DN when searching new targets and mechanisms on regulating inflammation in the development of DN. It is reported that TNF receptor related factor 6 (TRAF6) is an important protein on mediating TLR4 induced inflammatory response and plays a vital role in the development of several metabolic diseases. However, it is not clear whether TRAF6 involved in the development of DN. In our previous experiments, we found that TRAF6 inhibitor NBP2 attenuates STZ-induced DN, followed by inhibition of inflammatory response in vivo. In vitro, NBP2 inhibited high glucose-induced inflammatory response and cell injury. Thus, we hypothesis that TRAF6 plays an important role in DN and kidney inflammation and targeting inhibition of TRAF6 can attenuate the development of DN. We will elucidate the important effects of TRAF6 on high glucose-induced kidney inflammation and kidney injury in the levels of cellular, gene and animal through several molecular biology techniques, including gene silence, knockout, over-expression and RNA sequence. Meanwhile, we will illustrate the molecular mechanism of NBP2 on attenuating DN. This project will provide new target and drug in the treatment of DN.

慢性炎症是糖尿病肾病（DN）的重要病理过程之一，寻找DN过程中调控炎症的新机制和新靶点可能为DN防治提供新策略。TNF受体相关因子6（TRAF6）是TLR4激活炎症反应信号通路中的重要介导蛋白，已被发现在一些代谢性疾病发病过程中也有着重要作用，但TRAF6是否参与DN发生发展尚不清楚。我们前期实验发现，TRAF6抑制剂NBP2在小鼠体内能够缓解I型糖尿病肾病并伴随着炎症反应的下降，在体外NBP2能够抑制高糖诱导的炎症反应和细胞损伤。因此，我们假设TRAF6在DN和糖尿病肾炎中起重要介导作用，抑制TRAF6能够缓解DN的发生发展。本项目拟利用基因沉默、基因敲除、高表达、转录组测序等多种分子生物学技术，在细胞、基因和动物三个层面阐明TRAF6在高血糖诱导肾脏炎症和肾损伤中的重要作用，证实靶向TRAF6的小分子抑制剂能够缓解DN。本项目的实施将为DN的防治提供新的靶点和候选药物。

炎症在糖尿病肾病的发病机制中起着重要作用。TNF受体相关因子6（TRAF6）在toll样受体4和NF-κB信号传导过程中起重要作用。山奈酚是一种具有多功能药理财产的天然产物。在本项目中，我们研究了山奈酚是否能够减轻糖尿病肾病中的炎症反应。我们的数据显示，山奈酚显著降低了糖尿病小鼠的肾脏炎症、纤维化和肾功能障碍。这些效应与糖尿病小鼠肾脏中TRAF6的下调有关，表明TRAF6的潜在作用。通过AAV2-shTRAF6在小鼠中敲除TRAF6证实了TRAF6在糖尿病肾病中的重要性。在体外，用山奈酚处理NRK-52E细胞减弱了高糖诱导的炎症反应，与TRAF6表达下调相关。这一结论在NRK-52E细胞中通过敲低TRAF6的表达和过度表达得到进一步证实。我们的研究结果为TRAF6介导糖尿病引起的炎症导致肾功能障碍提供了直接证据。为糖尿病肾病的治疗提供了新的治疗方向和潜在治疗药物。