钙调激酶Ⅱ参与帕金森病运动并发症发病机制的实验研究  

A experimental research on the involvement of CaMKⅡ in the pathogenesis of motor complications in Parkinson＇s disease

作　　者：丁喜晴 朱忠方 崔桂云[1] 沈霞[1] 杨新新[1] 赵辉[2] 

机构地区：[1]徐州医学院附属医院神经内科,江苏徐州221004 [2]徐州市中心医院神经内科,江苏徐州221009

出　　处：《徐州医学院学报》2015年第7期427-431,共5页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金（81301077）;徐州市社会发展科技计划基金（XM138058）

摘　　要：目的观察钙调激酶Ⅱ（CaMKⅡ）抑制剂KN-93对帕金森病（PD）小鼠运动症状及异动症发生的影响并探讨其机制。方法将6-羟基多巴立体定向注射于小鼠纹状体制备PD小鼠模型，将造模成功的PD小鼠随机分为PD组、KN-92组和KN-93组，并设假手术组，每组16只。KN-92组和KN-93组给予左旋多巴（15ms／kg）和苄丝肼（12ms／kg）腹腔注射，假手术组及PD组腹腔注射等量生理盐水，每天1次，持续14天。在第11、12天左旋多巴注射前，KN-92组给予KN-92处理，KN-93组给予KN-93处理（2μl，浓度0．1s／L），假手术组及PD组给予等量生理盐水。于左旋多巴给药的第1、3、5、8、10、13天对各组大鼠进行异常不自主运动（AIM）评分，并于第0、4、9、14天进行圆柱体实验评分。此外，采用Western blot法检测纹状体区CaMKli及PKA磷酸化水平。结果随左旋多巴治疗时间的延长，大鼠AIM评分逐渐升高，给予KN-93处理后，AIM评分则明显下降（P〈0．01）；圆柱体实验提示给予左旋多巴后，KN-93组、KN-92组左前肢使用率较PD组明显升高（均P〈0．01）。Westernblot结果显示PD组及KN-92组CaMKlI及PKA磷酸化水平较假手术组明显升高（均P〈0．01），KN-93组CaMKⅡ及PKA磷酸化水平较KN-92组明显降低（均P〈0．01）。结论CaMKⅡ抑制剂KN-93可以减少PD小鼠异动症的发生而不影响左旋多巴的抗帕金森病作用。其机制可能是通过抑制PKA信号通路从而减少异动症的发生，提示抑制CaMKⅡ信号可能有助于减少PD小鼠异动症的发生。Objective To investigate the effects of a Ca2＋/calmodulin - dependent protein kinase Ⅱ （ CaMK Ⅱ ） inhibitor KN -93 on the motor deficits and dyskinesia in mice induced by 6 -hydroxydopamine （6 -OHDA） and potential mechanisms. Methods A Parkinson disease （PD） model was established in mice through stereotaxical injection of 6 - OHDA into the striatum. Then these mice were randomly divided into a PD group, a KN - 92 group, and a KN - 93 group （n = 16 ）. Meanwhile, another sixteen mice were subjected to sham -operation and set as controls. Mice in Groups KN -92 and KN- 93 were intraperitoneally administrated with 15 mg/kg of levodopa （L - DOPA） and 12 mg/kg of benserazide once per day for 14 days. On Days 11 and 12, they were further intrastriatally injected with KN -92 and KN -93 （2μ1, at a final concentration of 0.1 g/L） , respectively. In contrast, those in Groups PD and sham operation were intraperitoneally given equal volumes of normal saline. Then, the scores of abnormal involuntary movement （AIM） were estimated on Days 1, 3, 5, 8, 10 and 13, while the cylinder test was performed on Days 0, 4, 9 and 14. Furthermore, the levels of p - CaMKⅡ and p - PKA were measured by Western blot. Results The score of AIM was gradually in- creased along with the administration of L - DOPA, which was then remarkably decreased after KN - 93 treatment （ P 〈 0.01 ）. According to the results of the cylinder test, mice in both Groups KN- 93 and KN- 92 used the left forepaw more frequently than those in Group PD （ both P 〈 0.01 ）. Western blot analysis indicated that the levels of p - CaMK Ⅱ and p - PKA in Groups PD and KN - 92 were significantly higher than those in the sham operated group （ both P 〈 0.01 ）, while Group KN - 93 produced a markedly decreased level of p - CaMK Ⅱ and p - PKA than Group KN - 92 （ P 〈 0.01 ）. Conclusion The CaMKⅡ inhibitor KN - 93 can reduce the incidence of dyskinesia in PD mice, while not influencing the effects of L - DOPA against PD

关 键 词：帕金森病 运动并发症 磷酸化 钙调激酶Ⅱ KN-93 

分 类 号：R742[医药卫生—神经病学与精神病学]