Guo Q M, Liu J S
Department of Pharmacology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China.
Yao Xue Xue Bao. 2001 Sep;36(9):652-6.
To observe the change of Ca2+/calmodulin dependent protein kinase II (CaMK II) signal pathway in opioid dependent NG108-15 cells.
NG108-15 cells were used as an in vitro model system. Competitive protein binding assay and radioimmunoassay were used to examine the intracellular cAMP accumulation. Calmodulin activity was assayed by PDE method. CaMK II activity was assayed by gamma-32 P incorporation of syntide-2.
DPDPE long-term treatment increased calmodulin activity and CaMK II activity in both cytoplasm and nucleus of NG108-15 cells. Specific calmodulin antagonist W-7 was found to significantly inhibit the elevation of calmodulin and CaMK II activity which resulted from DPDPE long-term treatment, and CaMK II inhibitor KN-62 also inhibited elevation of CaMK II activity by DPDPE long-term treatment. When naloxone was added to NG108-15 cells which were long-term treated by DPDPE, calmodulin and CaMK II activity increased, indicating that naloxone withdrawal can increase Ca2+/CaMK II pathway activity.
The results indicate that Ca2+/CaMK II pathway was involved in the mechanisms of opioids dependence when DPDPE was long-term administered to NG108-15 cells.
观察阿片类药物依赖的NG108 - 15细胞中Ca2+/钙调蛋白依赖性蛋白激酶II（CaMK II）信号通路的变化。
以NG108 - 15细胞作为体外模型系统。采用竞争性蛋白结合试验和放射免疫分析法检测细胞内cAMP的积累。通过磷酸二酯酶（PDE）法检测钙调蛋白活性。通过syntide - 2的γ-32P掺入法检测CaMK II活性。
DPDPE长期处理增加了NG108 - 15细胞胞质和细胞核中的钙调蛋白活性和CaMK II活性。发现特异性钙调蛋白拮抗剂W - 7可显著抑制DPDPE长期处理导致的钙调蛋白和CaMK II活性升高，CaMK II抑制剂KN - 62也可抑制DPDPE长期处理引起的CaMK II活性升高。当向经DPDPE长期处理的NG108 - 15细胞中加入纳洛酮时，钙调蛋白和CaMK II活性增加，表明纳洛酮戒断可增加Ca2+/CaMK II信号通路活性。
结果表明，当对NG108 - 15细胞长期给予DPDPE时，Ca2+/CaMK II信号通路参与了阿片类药物依赖的机制。
