【作者】 贾静； 何芳； 谭琰； 宋晨晨； 王雅蕾； 万琴； 王雪； 徐自慧； 郭新志； 华茜；

【Author】 JIA Jing;HE Fang;TAN Yan;SONG Chenchen;WANG Yalei;WANG Xue;XU Zihui;WAN Liangqin;GUO Xinzhi;HUA Qian;College of Traditional Chinese Medicine, Beijing University of Traditional Chinese Medicine;College of Life Sciences,Beijing University of Traditional Chinese Medicine;Shanxi Cerebral Palsy Rehabilitation Hospital;

【机构】 北京中医药大学中医学院； 北京中医药大学生命科学学院； 山西省脑瘫康复医院；

【摘要】 研究背景:我们想要通过基因总片筛查CP的疾病基因表达量变化研究方法:1、CP患者基因表达谱的研究。招募先天性脑瘫、后天性脑瘫及正常儿童。采集所有研究对象的外周血,使用基因芯片进行扫描并分析找出与CP疾病相关的m RNA表达谱。2、天麻素对于CP的疗效评价及作用靶点。通过分析CP患者的临床资料,统计中药天麻素穴位注射治疗患者的内收肌角、粗大运动功能评分(GMFM-66)及精细运动功能评分(FMFM-45)的改变;并通过检测天麻素作用于M1型巨噬细胞的基因表达差异,确定天麻素在巨噬细胞参与炎性反应的关键作用靶点为BCL6基因。3、BCL6基因对天麻素作用于巨噬细胞的影响。通过观察天麻素对于M1及M2型巨噬细胞的作用,及敲除BCL6基因后该作用的变化,探讨其发挥抗炎抗凋亡作用的机制。研究结果:1.和正常对照组相比,CP病人组(无论先天CP还是后天CP组)有1008个基因表达有差异,其中214个基因表达升高,794个基因表达降低。2.通过对比CP患者天麻素穴位注射组和生理盐水穴位注射组治疗前后,以及天麻素穴位注射CP组及非脑瘫型神经疾病组治疗前后临床指标的变化,证实天麻素穴位注射对于缓解CP患者的内收肌高张力,提高GMFM-66以及FMFM-45评分有改善效果,且疗效显著优于非脑瘫型神经疾病患者;进而通过巨噬细胞实验,发现天麻素可以调节参与炎症反应为主的M1型巨噬细胞的BCL6基因表达上升。3.天麻素作用于M1型巨噬细胞使其BCL6基因表达量升高,对M2型则无此作用。此外天麻素可诱导M1型巨噬细胞分化为M2型,同时使受到氧化应激刺激巨噬细胞的细胞活力升高以及凋亡程度降低,但当敲除BCL6基因后,该诱导作用和调控细胞活性和细胞凋亡的能力消失。结论:1.与正常对照组相比,先天性及后天性脑瘫表达异常的基因与炎症、缺血缺氧、血管新生、细胞増殖及血管完整性的维持有关,提示脑瘫的病理机制确实与炎性反应和缺血缺氧反应共同相互作用有很大关联性。2.天麻素穴位注射对于缓解脑瘫疾病内收肌角、提高GMFM-66及FMFM-45评分的疗效显著,且明显高于非脑瘫型患者,提示天麻素发挥作用的机制与脑瘫的发病机制有共同通路或靶点。3.天麻素可诱导巨噬细胞由炎性反应为主的M1型分化为抗炎促修复为主的M2型,可以显著提高受到氧化应激刺激的巨噬细胞的活力,和降低其细胞凋亡的程度,且该作用是通过提高巨噬细胞的BCL6基因表达来介导。更多还原

【Abstract】 Background:we want to use themicroarray technology to screen the changes of the gene expression of CP. Methods: 1. m RNA expression profiles of CP Patients. We recruited congenital cerebral palsy,acquired cerebral palsy and normal children. The peripheral blood of all participants were collected and gene chip was used to scan and analyze the expression profiles of CP-related genes. 2. Effect of Gastrodin on CP and the related action target. The changes of adductor angle, gross motor function score(GMFM-66 and fine motor function score(FMFM-45) were measured by analyzing the clinical data of CP patients. Real time-PCR was used detect the difference of gene expression induced by LPS and differentiated into M1 type macrophage, and then determine the role of Gastrodin in macrophage involved in inflammatory response. 3. The effect of BCL6 gene on Gastrodin acting on macrophages.By observing the effects of Gastrodin on M1 and M2 macrophages and the changes of BCL6 gene knockout, the mechanism of Gastrodin exerting anti-inflammatory and anti-apoptotic effects was explored. Results: 1. Compared with the normal control group,there were 1008 differentially expressed genes in the CP patients group(whether congenital or acquired), of which 214 genes increased and 794 genes decreased. 2. By comparing the changes of clinical indexes before and after the treatment of Gastrodin acupoint injection group and saline acupoint injection group, as well as before and after the treatment of gastrodin acupoint injection group and non-cerebral palsy neurological disease group, it is confirmed that Gastrodin acupoint injection can alleviate the high tension of adductor muscle, improve the scores of GMFM-66 and FMFMFM-45 in patients with CP,and the curative effect is significantly better than that of non-cerebral palsy neurological disease group. Patient with disease; then through macrophage experiment, Gastrodin can regulate the increase of BCL6 gene expression in M1 macrophages,which are mainly involved in inflammatory reaction. 3. Gastrodin increased the expression of BCL6 gene in M1 macrophages,but had no effect on M2 macrophages. In addition, Gastrodin can induce M1 macrophages to differentiate into M2, and increase cell viability and decrease apoptotic degree of macrophages stimulated by oxidative stress. However, when BCL6 gene is knocked out, the induction and the ability to regulate cell activity and apoptosis disappear. Conclusion: 1.Compared with the normal control group, the abnormal expression of genes incongenital and acquired cerebral palsy is related to inflammation, ischemia and hypoxia, angiogenesis, cell proliferation and maintenance of vascular integrity,suggesting that the pathological mechanism of cerebral palsy is indeed closely related to the interaction between inflammatory response and ischemia and hypoxia response. 2.The changes of clinical indexes before and after treatment were compared with those of CP group and non-CP group. The results showed that the acupuncture point injection of Gastrodin in the patients with CP improved adductor angle release and GMFM-66 and FMFM-45 scores,and the curative effect was significantly better than that of non-CP neurological disease patients. Furthermore, macrophage experiment showed that Gastrodin could be adjusted to participate in.更多还原