摘要
目的:观察蛋白激酶A拮抗剂H-89对骨癌痛大鼠疼痛行为和背根神经节环磷腺苷效应元件结合蛋白(c AMP-response element binding protein,CREB)磷酸化水平的影响。方法:1检测骨癌痛大鼠背根神经节CREB磷酸化水平:将32只雌性SD大鼠随机分为假手术组和骨癌痛组(n=16),分组后在大鼠右侧胫骨髓腔内分别注射PBS或乳腺癌Walker 256细胞,肿瘤细胞接种后1,3,7,10,14 d检测大鼠机械痛阈,第14天提取骨癌痛组大鼠第2至第5腰椎右侧的背根神经节,应用免疫印迹法检测磷酸化CREB(p-CREB)的表达量。2检测H-89对大鼠疼痛行为及CREB磷酸化的影响:将32只骨癌大鼠随机均分为DMSO组和H-89组。在肿瘤细胞接种后的第9天至第14天每天鞘内注射DMSO或H-89。分别在给药前、给药后15,30,45和60 min进行疼痛行为测试。在第14天最后一次鞘内给药1 h后,快速提取大鼠右侧背根神经节,应用免疫印迹法检测p-CREB的表达量。结果:与假手术组比较,骨癌痛组大鼠机械痛阈明显降低,背根神经节中CREB磷酸化水平升高(P<0.05)。鞘内注射DMSO或H-89后,H-89组大鼠机械痛阈较DMSO组明显升高。同时H-89组CREB磷酸化水平较DMSO组明显降低(P<0.01)。结论:蛋白激酶A通过CREB磷酸化加剧大鼠骨癌痛。
Objective: To investigate the phosphorylation level of cyclic AMP response element binding protein( CREB) and evaluate the effects of intrathecal treatment with H-89,a protein kinase A( PKA) inhibitor,on dorsal root ganglion( DRG) CREB phosphorylation and nociceptive behavior in the rats with bone cancer pain. Methods: Thirty-two normal female Sprague-Dawley rats weighing 180- 200 g were randomly divided into 2 groups( n = 16 for each group) : sham operation group,bone cancer pain( BCP)group. The rat model of bone cancer pain was induced by injection of breast cancer cells( Walker 256) into the right proximal tibia bone marrow cavity. Sham operation group was injected by PBS. Mechanical pain threshold was measured at day 1,3,7,10,14 after inoculation. At day 14 after inoculation,the right side lumbar 2-5 dorsal root ganglion were harvested to detect the expression of phosphorylated CREB. Mechanical hyperalgesia was measured before inoculation and on day 1,3,7,10,14 after inoculation. After the behavioral test on day 14,the right lumbar 2- 5 DRG were removed to measure phosphorylated CREB with Western blotting. Thirty-two rats with bone cancer pain were randomly assigned to DMSO or H-89 group( n= 16 per group). On the 9th day after inoculation,intrathecal administration of DMSO or H-89 was given once daily until 14 d after inoculation. Mechanical pain threshold was measured before intrathecal injection and at 15,30,45 min and 1 h after intrathecal injection. The right side lumbar 2- 5 DRG was harvested af-ter the last injection. Western blotting analysis was used to examine the levels of p-CREB protein in different groups. Results: Compared with sham operation group,the mechanical pain threshold was significantly decreased in bone cancer pain group. The expression of p-CREB proteins was significantly higher in BCP rats than that of sham operation group rats( P < 0. 05). Compared with the DMSO group,mechanical pain threshold increased dramatically in H-89 group. The mechanical pain threshold of H-89 group reached peak at 15 min after drug injection and returned to baseline level at 1 h. The expression of p-CREB in H-89 group was much lower than the DMSO group( P < 0. 01). Conclusion: Protein kinase A may be involved in the development of bone cancer pain in rats through increasing the phosphorylation level of CREB in DRGs.
出处
《江苏大学学报（医学版）》
CAS
2015年第2期114-118,共5页
Journal of Jiangsu University：Medicine Edition
关键词
蛋白激酶A
骨癌痛
背根神经节
环磷腺苷效应元件结合蛋白
protein kinase A
bone cancer pain
dorsal root ganglion
cyclic AMP response element binding protein
