Akt通路及细胞凋亡在瘦素介导的大鼠慢性吗啡镇痛耐受中的作用  

Roles of Akt pathway and cell apoptosis in leptin-mediated chronic morphine antinociceptive tolerance

作　　者：胡芬[1] 崔宇[2] 郭瑞鲜[2] 莫利求[3] 冯鉴强[2] 

机构地区：[1]中山大学附属第一医院病理学教研室,广东广州510080 [2]中山大学中山医学院生理学教研室,广东广州510080 [3]中山大学附属第一医院黄埔院区麻醉科,广东广州510700

出　　处：《中国病理生理杂志》2016年第12期2163-2167,共5页Chinese Journal of Pathophysiology

基　　金：广东省医学科学技术研究基金资助项目(No.A2015415)

摘　　要：目的:探讨Akt(又称蛋白激酶B)及活化的caspase-3在瘦素介导的大鼠慢性吗啡镇痛耐受中的作用。方法:在SD大鼠建立慢性吗啡镇痛耐受模型;采用Western blotting法测定脊髓Akt和cleaved caspase-3的水平;免疫组织化学染色法检测脊髓磷酸化Akt(p-Akt)及cleaved caspase-3阳性细胞;免疫双染检测p-Akt及cleaved caspase-3阳性细胞的定位。结果:鞘内慢性注射吗啡(15μg)7 d可明显上调大鼠脊髓p-Akt及cleaved caspase-3的蛋白水平;在注射吗啡前30 min,鞘内注射瘦素拮抗剂(3μg)7 d能显著地抑制慢性吗啡处理对p-Akt和cleaved caspase-3的上调作用;p-Akt只定位在脊髓神经元,而cleaved caspase-3仅定位在星形胶质细胞;瘦素拮抗剂、Akt抑制剂及caspase-3抑制剂均能抑制慢性吗啡镇痛耐受。结论:脊髓Akt通路及活化的caspase-3参与瘦素介导的大鼠慢性吗啡镇痛耐受。AIM：To explore the roles of Akt（ also called protein kinase B） and active caspase-3 in the leptinmediated chronic morphine antinociceptive tolerance in rats.METHODS：A model of chronic morphine antinociceptive tolerance was established in the SD rats.The protein levels of spinal Akt and cleaved caspase-3 were tested by Western blotting.The technique of immunohistochemical staining was used to detect the immunoreactivity positive cells of phosphorylated（ p）-Akt and cleaved caspase-3 in the spinal cord.Double staining of immunohistochemistry was used to examine the cellular location of the p-Akt and cleaved caspase-3 positive cells.RESULTS：The chronic intrathecal injection of morphine（ 15 μg） for 7 d markedly upregulated the spinal protein levels of p-Akt and cleaved caspase-3 in the rats.Thirty min before injection of morphine,intrathecal injection of leptin antagonist（ 3 μg） for 7 d significantly attenuated the upregulation of the protein levels of p-Akt and cleaved caspase-3 induced by chronic morphine treatment.The p-Akt was exclusively observed in the spinal neurons.The cleaved caspase-3 was only localized with the spinal astrocytes.Intrathecal injecting the inhibitors of leptin,Akt and caspase-3 ameliorated the chronic antinociceptive tolerance.CONCLUSION：The spinal Akt pathway and active caspase-3 are involved in the leptin-mediated chronic morphine antinociceptive tolerance in rats.

关 键 词：瘦素 AKT CASPASE-3 吗啡镇痛耐受 

分 类 号：R363[医药卫生—病理学]