健脾益气法治疗肌萎缩侧索硬化症的作用机制研究

Amyotrophic lateral sclerosis (ALS) is a relatively rare, adult-onset, rapidly progressive and fatal disease that involves degeneration of spinal cord motor neurons.Although disease progression varies between individuals, toward the end stages of disease, most patients require ventilator support. Individuals with ALS most commonly die of respiratory failure or pneumonia within 2-5 y of diagnosis. There are no current treatments for ALS. Use ADAR2 flox/flox mice (AR2) which remained part of motor functions by transgenic methods to decribe the machanism for the onset of amyotrophic lateral sclerosis (ALS).The Jiawei Sijunzi decoction which can invigorate spleen and supplement vitality,has been proved in using for ALS patients and also has been found could improve the motor disorder of AR2 in pilot research,it might modify GluR2 Q/R or it's ADAR enzyme and had functions for neuron protection, and delayed the development of ALS,but the molecule mechanism has not yet been understand.In this study,we will investigate the neuroprotectional function of Jiawei Sijunzi decoction by AR2 mice model, and find its functional locations in molecule level by GluR2,ADAR2 enzyme and the edit efficiency of RNA.We aim to find molecule level evidence to suport in using of Jiawei Sijunzi decoction to treat ALS.

肌萎缩侧索硬化症（ALS）通常2-5年死于呼吸衰竭，目前尚缺乏有效防治手段。申请者前期通过ADAR2flox/+/VAChT-Cre.Fast杂合鼠产生ADAR2flox/flox/ VAChT-Cre.Fast (AR2)ALS小鼠模型，发现AR2脊髓前角细胞GluR2Q/R编辑率和ADAR2酶活性均与ALS运动功能减退和死亡率相关。研究发现中药健脾益气法能延缓AR2的运动功能减退。与临床使用此法改善ALS患者运动功能，延长生存率相当一致，其分子机制尚待阐明。本课题提出"健脾益气法能够从上游减少AR2小鼠运动神经元（MN）变性，提高AR2的GluR2Q/R编辑率与改善ADAR2酶的活性，减少MN变性，保护MN，延缓ALS的进展，提高生存率"的假说，观察其减少MN变性数量，改善运动功能、神经微观结构、Q/R编辑率、ADAR2酶的变化及对MN特异靶点的作用，揭示健脾益气法治疗ALS的分子机制

申请者前期通过ADAR2flox/+/VAChT-Cre.Fast杂合鼠产生ADAR2flox/flox/ VAChT-Cre.Fast (AR ALS小鼠模型,发现AR2脊髓前角细胞GluR2Q/R编辑率和ADAR2酶活性均与ALS运动功能减退和死亡率相关。通过日本改良日本引进的ADAR2flox/+/VAChT-Cre.Fast杂合鼠产生ADAR2flox/flox/ VAChT-Cre.Fast 成功地制作了能够体现散发性ALS进展以及解释其病理进展的小鼠模型。使用健脾益气法加味四君子汤能够从上游减少AR2小鼠运动神经元(MN)变性,提高AR2的GluR2Q/R编辑率与改善ADAR 2酶的活性,减少MN变性,保护MN,延缓ALS的进展,提高生存率；观察到加味四君子汤能够减少MN变性数量，改善运动功能、神经微观结构、Q/R编辑率、ADAR2酶的变化及对MN特异靶点的作用,揭示健脾益气法治疗ALS的分子机制。