目的:探索苁蓉舒痉颗粒对帕金森病斑马鱼的治疗效果及其分子机制,为苁蓉舒痉颗粒的药效学研究提供体内实验基础,以挖掘其临床价值.方法:将受精后6 h的斑马鱼胚胎随机分为对照组(1X E3养殖水处理)、模型组(750 μmol 6-OHDA溶液处理)和药物组(25 μg/mL苁蓉舒痉颗粒溶液处理).从受精后 0～120 h期间,每 24 h检测各组斑马鱼胚胎/鱼苗出膜率、存活率,120 h时在显微镜下观察各组斑马鱼的形态发育情况,利用行为学方法对斑马鱼进行触尾实验及实时荧光定量聚合酶链式反应检测多巴胺信号通路相关基因mRNA表达的变化.结果:苁蓉舒痉颗粒能显著提高帕金森病斑马鱼的存活率(P<0.01),并减少其畸形发生率(如卵黄吸收延迟、鱼鳔发育迟缓、脊柱弯曲、心包水肿等);有效减轻多巴胺神经元的受损程度,保护大脑形态的正常发育,挽救帕金森病斑马鱼自主运动能力(P<0.05);有效提高多巴胺相关基因的转录水平,其中dat和drd2a基因的表达水平有显著提高(P<0.05),对多巴胺神经元损伤导致的帕金森病斑马鱼有一定的保护作用.结论:苁蓉舒痉颗粒可能通过调节多巴胺相关基因dat和drd2a的表达,减少多巴胺神经元的凋亡,从而提高帕金森病斑马鱼的行为反应能力和存活率,对帕金森病的症状有显著的改善作用.
目的:利用七氯对斑马鱼多巴胺神经元的影响,建立斑马鱼帕金森病(PD)模型.方法:将斑马鱼鱼卵分为正常对照组(不予干预),750μmol/L 6-羟基多巴胺(6-OHDA)组和不同浓度七氯组(1 ng/mL、10 ng/mL、100 ng/mL、1000 ng/mL、10000 ng/mL),分别恒温孵育,每24 h观察胚胎发育情况并记录直到受精后120 hpf(h post fertilization,hpf),在120 hpf做行为学测定,然后实时荧光定量PCR(RT-qPCR)测定多巴胺通路酪氨酸羟化酶(TH2)的相对表达量.结果:与正常对照组比较,6-OH-DA组幼体出现发育迟缓、脊柱弯曲,运动距离缩短(P<0.05),TH2基因表达降低(P<0.05);与正常对照组比较,1～100 ng/mL七氯组幼体发育无明显变化,但运动距离缩短(P<0.05),TH2基因表达降低(P<0.05),1000～10000 ng/mL七氯组出现色素沉着分布和脊柱弯曲,而且运动距离更短(P<0.05)、TH2基因表达更低(P<0.05),变化具有浓度依赖性;与6-OHDA组比较,1000～10000 ng/mL七氯组运动距离更短,TH2基因表达更低,其中10000 ng/mL七氯组运动距离缩短和TH2基因表达降低有统计学差异(P<0.05).结论:传统PD模型造模药物6-OHDA和1000～10000 ng/mL七氯都可通过影响多巴胺神经元功能成功建立斑马鱼PD模型,其中10000 ng/mL七氯的造模效果比6-OHDA更显著.
Cerebral small vessel disease (CSVD) is a senile brain lesion caused by the abnormal structure and function of arterioles, venules and capillaries in the aging brain. The etiology of CSVD is complex, and disease is often asymptomatic in its early stages. However, as CSVD develops, brain disorders may occur, such as stroke, cognitive dysfunction, dyskinesia and mood disorders, and heart, kidney, eye and systemic disorders. As the population continues to age, the burden of CSVD is increasing. Moreover, there is an urgent need for better screening methods and diagnostic markers for CSVD, in addition to preventive and asymptomatic- and mild-stage treatments. Integrative medicine (IM), which combines the holistic concepts and syndrome differentiations of Chinese medicine with modern medical perspectives, has unique advantages for the prevention and treatment of CSVD. In this review, we summarize the biological markers, ultrasound and imaging features, disease-related genes and risk factors relevant to CSVD diagnosis and screening. Furthermore, we discuss IM-based CSVD prevention and treatment strategies to stimulate further research in this field.
