CAJ | 학술논문

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（15 mg／kg）和苄丝肼（12 mg／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111、2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0．1 g／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CaMKⅡ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）。 Western blot结果显示PD组及KN－92组CaMKⅡ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。
목적：관찰개조격매Ⅱ（ CaMKⅡ）억제제KN－93대파금삼병（ PD）소서운동증상급이동증발생적영향병탐토기궤제。방법장6－간기다파입체정향주사우소서문상체제비PD소서모형，장조모성공적PD소서수궤분위PD조、KN－92조화KN－93조，병설가수술조，매조16지。 KN－92조화KN－93조급여좌선다파（15 mg／kg）화변사정（12 mg／kg）복강주사，가수술조급PD조복강주사등량생리염수，매천1차，지속14천。재제111、2천좌선다파주사전，KN－92조급여KN－92처리，KN－93조급여KN－93처리（2μl，농도0．1 g／L），가수술조급PD조급여등량생리염수。우좌선다파급약적제1、3、5、8、10、13천대각조대서진행이상불자주운동（ AIM）평분，병우제0、4、9、14천진행원주체실험평분。차외，채용Western blot법검측문상체구CaMKⅡ급 PKA린산화수평。결과수좌선다파치료시간적연장，대서AIM평분축점승고，급여KN－93처리후，AIM평분칙명현하강（ P＜0．01）；원주체실험제시급여좌선다파후，KN－93조、KN－92조좌전지사용솔교PD조명현승고（균P＜0．01）。 Western blot결과현시PD조급KN－92조CaMKⅡ급PKA린산화수평교가수술조명현승고（균P＜0．01），KN－93조CaMKⅡ급PKA 린산화수평교KN－92조명현강저（균P＜0．01）。결론 CaMKⅡ억제제KN－93가이감소PD소서이동증적발생이불영향좌선다파적항파금삼병작용。기궤제가능시통과억제PKA신호통로종이감소이동증적발생，제시억제CaMKⅡ신호가능유조우감소PD소서이동증적발생。
Objective To investigate the effects of a Ca 2+/calmodulin -dependent protein kinase Ⅱ ( CaMKⅡ) inhibitor KN-93 on the motor deficits and dyskinesia in mice induced by 6-hydroxydopamine (6-OHDA) and poten-tial 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 (2μl, 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 -CaMKⅡand 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 -CaMKⅡand 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.This is probably related with the inhibition of the PKA signaling path-way.In other words, inhibition of CaMKⅡsignal may assist to reduce the incidence of dyskinesia in PD mice .