复方地黄颗粒调控MAPK/ERK信号通路治疗帕金森病的机制研究

Parkinson's disease (PD) can severely affect the patient’s quality of life and its incide is on the rise. Compound rehmannia granules (National Invention Patent) was developed on the basis of distinguished Chinese medical physicians and Chinese medicine literature. It has been proven effective for PD by clinical practice. To date, we’ve completed two NSFC (National Science Foundation of China) projects and one Shanghai Key Science & Technology Project regarding compound rehmannia granules. The laboratory and clinical study findings have suggested that the granules can significantly alleviate PD symptoms, possibly through regulating the MAPK/ERK signaling pathway. .This study adopted stereotaxic injections of 6-hydroxydopamine (6-OHDA) in the substania nigra of rats and 2-spot injection method for PD model preparation. The PD models were randomly allocated into a PD model group, PD98059 (ERK1/2 inhibitor) group, compound PD98059 group (compound rehmannia granules +PD98059) and compound rehmannia granules group, coupled with a normal control group and a sham-operation gorup. Aiming at the pathological role of MAPK signaling pathway in PD, we’ve conducted a comprehensive and dynamic observation on the action and mechanism of compound rehmannia granules for PD at different time frames (4 weeks and 6 weeks) in terms of functions, cells, proteins and molecular biology, with the goal of providing scientific basis for clinical PD treatment.

帕金森病(PD)的发病率日渐攀升，严重影响患者的生活质量。复方地黄颗粒（获国家发明专利）是挖掘名老中医经验及中医文献并经临床验证治疗PD有效的中药制剂。我们已经完成的2项国家自然基金项目和1项上海市科技攻关项目，从实验与临床方面证实了复方地黄颗粒对PD有明显的缓解作用，其作用可能与调控MAPK/ERK信号通路有关。.本研究采用经典的6-羟基多巴胺大鼠脑部黑质偏侧注射，并采用本课题组探索的2点法制备PD模型。PD模型随机分为PD模型组、PD98059组（PD98059为ERK1/2抑制剂）、复方PD98059组（复方地黄颗粒+PD98059）、复方地黄颗粒组（复方地黄颗粒），并设正常对照组、假手术组。针对PD发病的MAPK/ERK信号通路环节，每组在不同的时间点（4周、6周），分别从功能、细胞、蛋白、分子生物学角度，全面、动态揭示复方地黄颗粒治疗PD的作用及机制，以期为PD的临床治疗提供依据。

帕金森病(PD)的发病率日渐攀升，严重影响患者的生活质量。复方地黄颗粒是挖掘名老中医经验及中医典籍并经临床验证治疗PD有效的中药制剂。本研究采用经典的并经本课题组反复验证的6-羟基多巴胺大鼠脑部黑质偏侧2点法注射制备PD模型。将成功模型随机分为PD模型组、MAPK抑制剂组、复方地黄颗粒+抑制剂组、复方地黄颗粒组，并设正常对照组、假手术组。每组在4周、6周，分别从功能、细胞、蛋白、分子生物学角度，观察了复方地黄颗粒对MAPK信号通路的影响及其作用机制。结果发现，与正常对照组、假手术组比较，PD模型组大鼠黑质纹状体P-ERK1/2、p-CREB蛋白表达明显升高，TH、Nurr1、Lxm-1b、Pitx3蛋白、Nurr1 mRNA、Lmx-1b mRNA、Pitx3 mRNA、NGF mRNA、BDNF mRNA、GDNFmRNA表达明显降低，SOD、GSH、GSH-Px含量明显降低，MDA含量明显升高，Cleaved Caspase3、Caspase9、Cyt-c、p53、C-myc蛋白表达、TUNEL阳性细胞数明显升高，Bcl-2/Bax表达明显降低，p-JNK/ JNK、p-c-Jun/c-Jun、p-c-Fos/c-Fos表达明显升高，差异均有统计学意义（P﹤0.05，或P﹤0.01，或P﹤0.001）；复方地黄颗粒组可明显降低P-ERK1/2、p-CREB蛋白表达，升高TH、Nurr1、Lxm-1b、Pitx3蛋白、Nurr1 mRNA、Lmx-1b mRNA、Pitx3 mRNA、NGF mRNA、BDNF mRNA、GDNFmRNA表达，升高SOD、GSH、GSH-Px含量，降低MDA含量，降低Cleaved Caspase3、Caspase9、Cyt-c、p53、C-myc蛋白表达、TUNEL阳性细胞数，升高Bcl-2/Bax表达量，降低p-JNK/ JNK、p-c-Jun/c-Jun、p-c-Fos/c-Fos表达，差异有统计学意义（P﹤0.05，或P﹤0.01，或P﹤0.001），且随干预时间的延长，疗效更加明显。提示复方地黄颗粒通过调控MAPK/ERK信号通路，保护DA能神经元，促进神经元干细胞分化、增殖，调控MAPK/JNK信号通路，抗神经细胞凋亡，从而起到治疗PD的作用。