促肾上腺皮质激素释放激素受体1拮抗剂CP154526对大鼠海马神经元凋亡的影响  

Effects of CP154526,the corticotropin releasing hormone receptor 1 antagonist,on rat hippocampal neuron apoptosis

作　　者：周毅[1] 刘巍[2] 张悦[3] 

机构地区：[1]河北医科大学第二医院神经内科,河北石家庄050017 [2]河北医科大学病理学教研室,河北石家庄050017 [3]河北医科大学临床诊断学教研室,河北石家庄050017

出　　处：《中国药理学通报》2015年第4期504-509,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81302625);河北省高等学校科学技术研究优秀青年基金项目(No Y2012001)

摘　　要：目的观察促肾上腺皮质激素释放激素(CRH)受体1拮抗剂CP154526对海马神经元凋亡的影响。方法原代培养大鼠海马神经元,噻唑蓝(MTT)法测定细胞存活率,然后分为4组:正常对照组(Con)、CRH刺激组(CRH)、CRH和CP154526共同刺激组(CRH+CP)、CP154526刺激组(CP)。TUNEL、流式细胞术AnnexinⅤ-PI法检测神经元凋亡率;Western blot检测凋亡蛋白Bax、Bcl-2、caspase-3表达水平。结果 10-8mol·L-1的CRH作用于海马神经元后,细胞存活率下降(P<0.05);50 mmol·L-1的CP154526可明显提升神经元存活率(P<0.05)。与正常对照组相比,CRH刺激后神经元凋亡率增加,Bax/Bcl-2比值增高,caspase-3表达增加;加用CP154526可明显降低神经元凋亡率、Bax/Bcl-2比值及caspase-3表达水平;而单独应用CP154526对凋亡没有明显影响。结论一定浓度的CRH可诱导海马神经元细胞凋亡,其受体1拮抗剂CP154526可有效减轻细胞凋亡,发挥神经保护作用。Aim To investigate the effects of CP154526,a corticotropin releasing hormone（ CRH）receptor 1 antagonist,on the hippocampal neuron apoptosis. Methods Rat hippocampal neurons were primarily cultured. Cell viability was estimated usingMTT assays. Neurons were randomly divided into four groups： Normal cultures（ Control）; CRH-exposed cultures（ CRH）; CRH and CP154526 co-exposed cultures（ CRH ＋ CP）; CP154526-exposed cultures（ CP）. Cell apoptosis was examined by TUNEL or flowcytometry Annexin Ⅴ-PI staining. The protein levels of Bax,Bcl-2 and Caspase-3 were investigated by Western Blotting. Results 10- 8mol·L^-1CRH decreased cell viability of cultured hippocampal neuron（ P 〈0. 05）,while 50 mmol ·L^-1CP154526 significantly increased neuron viability（ P 〈0. 05）. Compared with Control group,cell apoptotic rate,the ratio of Bax and Bcl-2 and the protein level of Caspase-3 were elevated in hippocampal neuron induced by CRH. Combinedwith CP154526 reversed the effects of CRH. Application of CP154526 alone had no obvious effects on cell apoptosis. Conclusions A certain concentration of CRH can induce hippocampal neuron apoptosis,and its receptor 1 antagonist CP154526 can effectively reduce the apoptosis and play a neuroprotective role.

关 键 词：促肾上腺皮质激素释放激素 CP154526 海马 神经元 凋亡 原代培养 

分 类 号：R714.21[医药卫生—妇产科学]