糖痹康干预糖尿病周围神经病变内质网应激诱导细胞凋亡作用机制研究

Diabetic peripheral neuropathy (DPN) is one of the three major complications of diabetes and has a high incidence. And it has been the main cause of Mutilation. Chinese medicine has accumulated rich experience in the treatment of DPN, especially in the field of improving symptoms of DPN patients. But we still don’t know how it does work. Endoplasmic reticulum stress-induced apoptosis is the key mechanism of DPN development process. IRE1α-XBP1 pathway-induced apoptosis plays an important roles in relieving endoplasmic reticulum stress and apoptosis in the treatment of DPN. We have found that herbal compound Tang bi kang can alleviate inflammation，oxidative stress，endoplasmic reticulum stress and reduce apoptosis, repair peripheral nerve injury and improve the sciatic nerve conduction velocity. In this study, based on previous studies, we predict that Tang bi kang protects peripheral nerve function may through regulates IRE1α-XBP1 pathway to recover endoplasmic reticulum stress and reduce apoptosis. Now we want to set up animal experiment to study how Tang bi kang regulates IRE1α-XBP1 pathway to against endoplasmic reticulum stress and apoptosis, and provide a basis for its clinical practice.

糖尿病周围神经病变(DPN)是糖尿病三大并发症之一，临床发病率较高，是糖尿病患者致残首因。中医药在治疗DPN方面积累了丰富经验，尤其在改善DPN患者临床症状方面疗效显著，其科学内涵亟待阐明。内质网应激诱导细胞凋亡是DPN的关键发病机制。干预内质网应激IRE1α-XBP1通路诱导的细胞凋亡对治疗DPN有重要意义。前期研究发现，临床有效中药复方糖痹康（由黄芪桂枝五物汤化裁）具有抗炎、抗氧化、抗内质网应激并减少细胞凋亡造成的周围神经损伤，改善坐骨神经传导速度的作用。本研究在前期研究基础上，拟从内质网应激诱导细胞凋亡角度，通过动物实验，从整体、组织、分子水平观察糖痹康干预内质网应激诱导的细胞凋亡以及多靶点调节IRE1α-XBP1通路的作用机制，研究和探讨中药复方治疗DPN的作用机理，为中药复方糖痹康治疗DPN提供理论和实验依据，为研发治疗DPN的创新中药奠定基础。

项目背景：前期实验研究表明糖痹康具有抗炎抗氧化作用，糖痹康可能存在良好的改善机体抗内质网应激能力，为证实这一假说进行本实验研究。本研究通过是观察中药复方糖痹康对糖尿病周围神经病变内质网应激诱导细胞凋亡的影响，探讨中药复方治疗DPN 的作用机理，为研发治疗DPN 的创新中药奠定基础。. 主要内容：高脂饲料联合STZ 诱导DPN动物模型，糖痹康干预16周，α-硫辛酸作西药对照组，实验中记录大鼠一般状况，体重，血糖，给药结束后测定坐骨神经传导速度，取坐骨神经组织做病理切片，行HE，LBF快蓝染色观察坐骨神经形态变化，透视电镜观察坐骨神经超微结构。检测血清中内质网应激相关指标ROS和SOD的含量， Western blot检测坐骨神经组织中IRE1α，p-IRE1α，XBP1s，CHOP，Bcl-2，Bax，caspase12蛋白的表达，Real-time PCR检测坐骨神经组织中IRE1α，XBP1s，CHOP，Bcl-2，Bax，caspase12 mRNA的表达。. 重要结果：动物实验表明，中药复方糖痹康能降低有效DPN大鼠血糖，抑制体重减轻，提高坐骨神经传导速度，减轻神经组织病理学损害，上调SOD水平及Bcl-2表达，下调ROS水平及IRE1α，p-IRE1α，XBP1s，CHOP，Bax，caspase12表达。. 科学意义：中药复方糖痹康能有效治疗糖尿病周围神经病变，保护和修复坐骨神经损伤，减轻机体和组织细胞的损伤，可能与其调节 IRE1α- XBP1 通路，减少细胞凋亡，恢复机体内质网稳态有关。