中文摘要
注意力缺陷-多动综合征（ADHD）发病与中枢神经系统皮质-纹状体-丘脑（CSTC）环路多巴胺（DA）代谢异常相关。前期研究表明以钩藤为君药的宁动颗粒治疗ADHD安全有效，钩藤提取物钩藤碱（RP）具有改善ADHD小鼠症状，体外能增加多巴胺能神经元（DAN）代谢产物浓度。据此，提出科学假设：RP通过调节CSTC环路DA代谢而减轻ADHD症状。为验证假设，探讨RP调节DA代谢机制：本研究拟对RP在体外72小时作用于SiRNA-DAT干扰后的DAN、体内12周作用于多巴胺转运体（DAT）基因敲除的ADHD小鼠。通过体内、体外实验：每周监测小鼠行为学改变，分时间段检测DAN代谢酶（MAO/TH）活性、代谢产物（DOPAC/HVA）浓度、受体（DAR1/DAR2）、DAT蛋白及mRNA表达，揭示RP调节ADHD小鼠CSTC环路DA代谢机制。成果将为探索ADHD中医药替代疗法及RP新药研发提供实验依据。
英文摘要
Attention deficit/hyperactivity disorder (ADHD) is involved in the pathogenesis of the metabolic abnormalities of dopamine (DA) in cortico-striatal-thalamo-cortical(CSTC) circuits in brain. As a Traditional Chinese Medicine (TCM) prescription Ningdong granule (NDG) has proved to be effective on the treatment of ADHD for several years in China. Uncaria is a kind of key herb of NDG. Our previous studies have also indicated that the herbal uncaria and its main extract rhynchophylline (RP) could improve some symptoms of ADHD and regulate DA metabolism in brain. However, the working mechanism on regulating DA still remains obscure. Previously, we have developed an ADHD mouse model with DAT gene knockout. To investigate how RP regulate DA metabolism, we plan to utilize the ADHD model mice to monitor their behaviors for 12 weeks by RP treatment in vivo, and to test the enzymes MAO/TH that are involved in the DA metabolism, the metabolic product HVA and DOPAC, the DA receptors (DAR1 and DAR2) and the DA transporte in dopaminergic neurons of CSTC for 72h at different time point in vitro. These results will provide some valuable information for the development of RP, and may also lead to the establishment of an effective therapy in TCM for ADHD children.
结题摘要
注意力缺陷-多动综合征（ADHD）发病与中枢神经系统皮质-纹状体-丘脑（CSTC）环路多巴胺（DA）代谢异常相关。前期研究表明以钩藤为君药的宁动颗粒治疗ADHD安全有效，钩藤提取物钩藤碱（RP）具有改善ADHD小鼠症状，体外能增加多巴胺能神经元（DAN）代谢产物浓度。据此，提出科学假设：RP通过调节CSTC环路DA代谢而减轻ADHD症状。为验证假设，探讨RP调节DA代谢机制：本研究拟对RP在体外作用于多巴胺转运体（DAT）基因敲除的ADHD小鼠。通过体内、体外实验：每周监测小鼠行为学改变，分时间段检测DAN代谢酶（MAO/TH）活性、代谢产物（DOPAC/HVA）浓度、受体（DAR1/DAR2），揭示RP调节ADHD小鼠CSTC环路DA代谢机制。研究结果表明：RP能有效改善DAT基因敲除小鼠的记忆；DAT模型组中，在时间上3、6h作用多巴胺的代谢达到高峰；DA浓度与DAT模型组及健康对照组比较，DAT+钩藤碱组与DAT+宁动颗粒组培养液中含量均明显降低（P＜0. 05），HVA浓度、MAO浓度、BDNF浓度均显著增高，且DAT+钩藤碱组与DAT+宁动颗粒组间无显著性差异；LPS介导的炎性组中，在时间上3、6h作用TNF-α，IL-1β的含量达到高峰；TNF-α，IL-1β浓度，与LPS模型组比较，空白对照组、LPS+钩藤碱组与LPS+宁动颗粒组培养液中IL-1β含量均显著降低（P＜0. 05），且LPS+钩藤碱组与LPS+宁动颗粒组间无显著性差异。结果表明：钩藤碱能有效改善ADHD模型小鼠记忆能力，可以通过调节MAO活性及BDNF活性促进多巴胺能神经元代谢，也能显著抑制LPS诱导的神经元及神经胶质细胞的炎性因子TNF-α，IL-1β，起到抗炎而保护神经元作用。