脊髓Kalirin-7通过PSD95-NR2B在2型糖尿病神经病理性疼痛中的作用及机制

The World Health Organization estimates that the global prevalence of diabetes is currently approaching 5%，90% of all diabetic patients are diabetes type 2. Diabetic peripheral neuropathy is one of the most common consequences of diabetes and might be associated with diabetes type 1 and type 2 and the proposed therapies are inefficient. However, diabetic neuropathy is not always painful, almost 50% of all diabetic patients are affected with diabetic neuropathic pain (DNP).The pathogenesis of DNP is complicated, and the mechanism of this disease remains poorly understood. Emerging evidence suggested that NMDA receptor 2B subunit NR2B mediated central sensitization of neuropathic pain, and Kalirin-7 was supposed to involved in neuropathic pain. In the conditioning with high blood glucose, which mechanism induced the neuropathic pain? The NR2B of spinal cord suggested to play a crucial role in the produce and maintenance of pathological pain,our previous studies indicated that the expressions of phospholicated NR2B was increased in neurons of type 2 diabetic neuropathic pain rats in spinal cord, NR2B maybe involved in the pathological mechanisms of DNP.In the present project,diatetes rats with or without DNP will be the ke directly interacted with NR2B in neuropathic pain. But those main suppositiony point,and kalirin-7 will be supposed be the link with PSD95 and NR2B, animals with or without DNP will be used,the experiment of animal will be the first step to investigated the expression and location of kalirin-7,PSD95 and NR2B in neuron,microglia,astrocytes in spine .Then we will culture microglia, astrocytes, neuron and co-culture microglia or astrocyte with neuron to explain the effect of high glucose on the the expressions and interation of Kalirin-7, PSD95 and NR2B in spine of type II diabetic neuropathic pain in rats.At the end ,the inhibitions of kalirin-7 and NR2B using inhibitors or SiRNA technique will be used to explain the kalirin-7 via PSD95-NR2B play an important role in type 2 diabetic neuropathic pain rats. The implementation of this project will provide the theoretical and experimental basis for the clinic therapy of DNP.

糖尿病患者占总人口5%，其中90%为Ⅱ型糖尿病，有近50%的糖尿病患者伴有糖尿病神经病理性疼痛（DNP）。研究提示脊髓NR2B在中枢敏化和慢性痛的发生发展中具有重要作用。现有关于神经病理性疼痛产生机制的研究结果均为已发生病理性疼痛的动物和正常动物比较得出。我们前期研究提示，DNP大鼠脊髓和背根神经节NR2B的表达远高于正常大鼠，而NR1未见变化。进一步选取同样在2型糖尿病情况下，伴有和不伴有神经病理性疼痛大鼠，发现伴有神经病理性疼痛大鼠脊髓NR2B的表达明显高于不伴有疼痛的糖尿病大鼠。本实验将以糖尿病伴有或不伴有DNP为切入点，以Kalirin-7为纽带，选取血糖升高但痛阈变化明显或不明显的动物，通过整体动物-离体培养细胞-整体动物实验，阐明脊髓Kalirin-7通过PSD95-NR2B在DNP产生和维持中的作用及确切机制，为DNP的临床治疗提供实验基础和理论依据。

糖尿病患者占总人口5%，其中90%为Ⅱ型糖尿病，有近50%的糖尿病患者伴有糖尿病神经病理性疼痛（DNP）。研究提示脊髓NR2B在中枢敏化和慢性痛的发生发展中具有重要作用。本项目以糖尿病伴有或不伴有DNP为切入点，以Kalirin-7为纽带，选取血糖升高但痛阈变化明显或不明显的动物，通过整体动物-离体培养细胞-整体动物实验，发现：DNP大鼠脊髓Kalirin-7表达增强，在PDZ区和PSD95结合，引起NR2B磷酸化，NMDA受体功能增强，中枢敏化形成，最终导致DNP的发生；脊髓SGK1通过介导HDAC4磷酸化和HDAC4易位在异常性疼痛发展中起重要作用；SIRT1-CDK5-Kalirin7-p-NR2B信号通路参与Ⅱ型糖尿病神经病理性疼痛的形成与发展；ROS通过调节TXNIP-NLRP3-NR2B信号通路参与大鼠Ⅱ型糖尿病神经病理性疼痛。本研究阐明脊髓Kalirin-7通过PSD95-NR2B在DNP产生和维持中的作用及确切机制，为DNP的临床治疗提供实验基础和理论依据。