Zhu C Y, He C J, Yao M, Xu L S, An K, Liu Q Y, Chen Y J, He Q L, Huang B, Zhou X Y
Department of Anesthesiology, Bengbu Medical College, Bengbu 233030, China.
Zhonghua Yi Xue Za Zhi. 2018 Jan 23;98(4):289-293. doi: 10.3760/cma.j.issn.0376-2491.2018.04.010.
To investigate the relationship between C-C chemokine receptor type 2(CCR2) and P38 mitogen-activated protein kinase (P38MAPK) signaling pathway in the spinal cord of rats and further clarify the mechanism of bone cancer pain (BCP). A total of 92 healthy female SD rats, of which 60 were subjected to behavioral tests using a ciliary mechanical stimulation needle. SD rats were randomly divided into six groups: sham operation group (group S), bone cancer pain group (group B), sham operation + DMSO solvent group (group SD), bone cancer pain + DMSO solvent group (group BD), sham operation + RS102895 CCR2 inhibitor group (group SR), bone cancer pain + RS102895 CCR2 inhibitor group (group BR), and Von Frey was used in the behavioral test. Another 32 SD rats were randomly divided into the following 8 groups (n=4): sham operation group (group S), bone cancer pain 5 d group (group B5), bone cancer pain 9 d group (group B9), bone cancer pain 14 d group (group B14), bone cancer pain + DMSO solvent group (group BD), bone cancer pain + RS102895 CCR2 inhibitor 0.5 h group (group BR0.5 h), bone cancer pain + RS102895 CCR2 inhibitor 4 h group (group BR4 h), bone cancer pain + RS102895 CCR2 inhibitor 12 h group (group BR12 h). Western blot was used to detect the expression of P38, p-P38 and CCR2 in spinal cord of rats. At day 5, 7, 9, 14, 21 post-injection, mechanical withdrawal thresholds of group S were(30.9±1.5), (31.9±1.2), (32.0±1.1), (31.6±1.5), (32.2±1.4)g respectively, the mechanical withdrawal thresholds of group B were( 26.4±0.7), (24.4±0.8), (21.4±0.8), (13.5±0.4), (9.9±0.2)g respectively, the mechanical withdrawal thresholds in group B decreased obviously versus group S, and the differences were statistically significant(=-13.177, -16.660, -23.778, -35.574, -48.401, all <0.01). At day 9 post-injection, the mechanical withdrawal thresholds in SD, BD, SR and BR groups were (32.4±1.7), (19.4±1.1), (32.1±1.3), (26.3±1.0) g respectively, the difference was statistically significant (=224.681, <0.01), and the mechanical withdrawal thresholds in group BD decreased obviously versus group SD, while the mechanical withdrawal thresholds in group BR increased obviously versus group BD. The expression levels of p-P38 in spinal cord of group S, group B5, group B9 and group B14 were(0.08±0.03), (0.20±0.05), (0.40±0.17), (0.65±0.14)respectively, the expression levels of CCR2 were(0.08±0.04), (0.18±0.05), (0.30±0.09), (0.58±0.07)respectively, the difference was statistically significant(=19.123, 40.746, all <0.01), and the expression of p-P38 and CCR2 in group B9 were showed a significant up-regulation versus group S. The expression levels of p-P38 in spinal cord of group BD, group BR0.5 h, group BR4 h and group BR12 h were (0.57±0.06), (0.17±0.11), (0.03±0.01), (0.25±0.11)respectively, and the difference was statistically significant(=29.582, <0.01). The expression of p-P38 in group BR0.5 h, BR4 h, BR12 h showed a significant down-regulation versus group BD. CCR2 in the spinal cord may be involved in the development of bone cancer pain by activating P38MAPK signaling pathway in rats.
探讨大鼠脊髓中C-C趋化因子受体2（CCR2）与P38丝裂原活化蛋白激酶（P38MAPK）信号通路之间的关系，进一步阐明骨癌痛（BCP）的机制。选取92只健康雌性SD大鼠，其中60只采用纤毛机械刺激针进行行为学测试。将SD大鼠随机分为6组：假手术组（S组）、骨癌痛组（B组）、假手术+二甲基亚砜溶剂组（SD组）、骨癌痛+二甲基亚砜溶剂组（BD组）、假手术+RS102895 CCR2抑制剂组（SR组）、骨癌痛+RS102895 CCR2抑制剂组（BR组），行为学测试采用von Frey法。另取32只SD大鼠随机分为以下8组（n=4）：假手术组（S组）、骨癌痛5 d组（B5组）、骨癌痛9 d组（B9组）、骨癌痛14 d组（B14组）、骨癌痛+二甲基亚砜溶剂组（BD组）、骨癌痛+RS102895 CCR2抑制剂0.5 h组（BR0.5 h组）、骨癌痛+RS102895 CCR2抑制剂4 h组（BR4 h组）、骨癌痛+RS102895 CCR2抑制剂12 h组（BR12 h组）。采用蛋白质免疫印迹法检测大鼠脊髓中P38、p-P38及CCR2的表达。注射后第5、7、9、14、21天，S组机械缩足阈值分别为（30.9±1.5）、（31.9±1.2）、（32.0±1.1）、（31.6±1.5）、（32.2±1.4）g，B组机械缩足阈值分别为（26.4±0.7）、（24.4±0.8）、（21.4±0.）、（13.5±0.4）、（9.9±0.2）g，B组机械缩足阈值较S组明显降低，差异有统计学意义（t=-13.177、-16.660、-23.778、-35.574、-48.,均P<0.01）。注射后第9天，SD组、BD组、SR组和BR组机械缩足阈值分别为（32.4±1.7）、（19.4±1.1）、（32.1±1.3）、（26.3±1.0）g，差异有统计学意义（F=224.681，P<0.01），BD组机械缩足阈值较SD组明显降低，BR组机械缩足阈值较BD组明显升高。S组、B5组、B9组和B14组脊髓中p-P38表达水平分别为（0.08±0.03）、（0.20±0.05）、（0.40±0.17）、（0.65±0.14），CCR2表达水平分别为（0.08±0.04）、（0.18±0.05）、（0.30±0.09）、（0.58±0.07），差异有统计学意义（F=19.123、40.746，均P<0.01），B9组p-P38和CCR2表达较S组明显上调。BD组、BR0.5 h组、BR4 h组和BR12 h组脊髓中p-P38表达水平分别为（0.57±0.06）、（0.17±0.11）、（0.03±0.01）、（0.25±0.11），差异有统计学意义（F=29.582，P<0.01）。BR0.5 h组、BR4 h组、BR12 h组p-P38表达较BD组明显下调。脊髓中的CCR2可能通过激活大鼠P38MAPK信号通路参与骨癌痛的发生发展。
