临床神经病学杂志2017，Vol.30Issue(1)：40-44,5.
钙调激酶Ⅱ抑制剂在异动症发生中的分子机制研究
Study on the molecular mechanism of calmodulin kinase Ⅱ inhibitor on the occurrence of dyskinesia
摘要
Abstract
Objective To investigate the molecular mechanism of calmodulin kinase Ⅱ ( CaMK Ⅱ) inhibitor on the occurrence of dyskinesia. Methods 6-hydroxy dopamine was injected into the right medial forebrain bundle to establish the model of Parkinson's disease (PD) mice. The established PD mice were divided into PD group(n=11), KN-92 group(n=11) and KN-93 group(n=11). Another group of sham-operated mice was involved as control(n=10). KN-92 group and KN-93 group received intraperitoneal injection of L-DOPA (15 mg/kg) and Benserazide (12 mg/ kg) , mice in sham-operated group and PD group were injected with the same volume of physiological saline, once daily for 14 d. At 11 d and 12 d, KN-92 or KN-93 (2 μl, concentration 0. 1 μg/μl) was intrastriatumly injected into mice before L-doap treatment. Mice in sham -operated group and PD group received normal saline treatment. At 1 d, 3 d, 5 d, 8 d, 10 d and 13 d of treatment, abnormal involuntary movement scale ( AIM) was performed in sham -operated group, PD group,KN-92 group and KN-93 group. At 4 d, 9 d and 14 d of treatment,the rate of usage of left fore was evaluated. The levels of cAMP-response element binding protein ( CREB) , dopamine and cAMP-regulated phosphoprotein of Mr 32000 ( DARPP-32 ) , phosphorylated levels of CREB and DARPP-32 were determined by western blot. Results There was no difference in the rate of usage on left fore after operation when compared with original condition in sham-operated group. Surprisingly the rate of usage on left fore decreased clearly after treatment and there was significantly difference in using rate when compared with it before treatment in PD treatment group ( P<0. 01 ) . The rate of usage on left fore increased clearly after treatment and there was significantly difference in KN-92 and KN-93 treating group when compared with it before treatment ( P<0. 01 ) . Before treatment there was no difference in rate of usage on left fore among all groups. At 4 d and 9 d after treatment the rate of usage on left fore in PD, KN-92, and KN-93 group was lower than that in sham-operated group ( P<0. 01). At 14 d, there was no difference between KN-93 group and sham-operated group but in both PD and KN-92 treating group rate of usage still lower than that in sham-operated group (P<0. 01). The rate of usage on left fore at 4 d, 9 d and 14 d in KN-92 group and at 4 d and 9 d in KN-93 group were significantly higher than those in PD group in corresponding days ( all P<0. 01 ) . Especially at 14 d, the rate of usage on left fore in KN-93 group was significantly higher than that in KN-92 group (P<0. 01). The AIM score in mice increased gradually in 1 d, 3 d, 5 d, 8 d and 10 d after treatment with KN-92 and KN-93 when compared with PD treatment (P<0. 01). Except at 14 d, the AIM score was higher after KN-92 treatment there was no difference on all other observing days between KN-92 and KN-93 treatment. The phosphorylated levels of CREB and DARPP-32 were both higher in mice of KN-92 group than those in PD group, and the differences were statistically significant (P<0. 01). Moreover, the phosphorylated levels of CREB and DARPP-32 in mice of KN-93 group were significantly reduced, and the differences were statistically significant (P<0. 01). Conclusion L-DOPA induced dyskinesia may be associated with CaMKⅡand its inhibitor KN-93 may be used in the treatment of PD motor complications as a new option.关键词
帕金森病/运动并发症/磷酸化/钙调激酶Ⅱ/KN-93Key words
Parkinson's disease/motor complications/phosphorylation/calmodulin kinase Ⅱ/KN-93分类
医药卫生引用本文复制引用
朱忠方,丁喜晴,杨新新,崔桂云,花放,沈霞..钙调激酶Ⅱ抑制剂在异动症发生中的分子机制研究[J].临床神经病学杂志,2017,30(1):40-44,5.基金项目
国家自然科学基金(81301077) （81301077）
江苏省高校自然科学基金(13KJB320026) （13KJB320026）
