目的 观察鞘内注射内皮素A受体(ETAR)拮抗药对大鼠骨癌疼痛(BCP)改善作用及其对细胞外调节蛋白激酶(ERK)通路的影响。方法 取60只大鼠均进行鞘内置管,随机分为假手术(sham)组、假手术+ETAR拮抗药(sham+BQ123)组、骨癌痛(BCP)组、骨癌痛+ETAR拮抗药(BCP+BQ123)组。BCP组、BCP+BQ123组采用股骨远端骨髓腔内接种Walker256细胞法建立BCP大鼠模型,sham组、sham+NS同法注射等量生理盐水。建模成功大鼠于建模第14 天,BCP+BQ123组和sham+BQ123组各14只,鞘内注射7 μl BQ123;BCP组13只、sham组14只鞘内注射等量生理盐水。鞘内注射前即刻、注射后0.5、1.0、1.5、2.0、2.5、3.0 h分别评估各组大鼠疼痛行为学:机械性缩足反射阈值(PWT)、自发抬足次数(NSF);末次评估疼痛行为学后,影像学评估各组骨质破坏情况;RT-qPCR、Western blot法检测脊髓组织中内皮素1(ET1)、ETAR、ERK1/2 mRNA和蛋白表达量及p-ERK1/2蛋白表达量。结果 与sham组、sham+BQ123组比较,注射前BCP组、BCP+BQ123组的PWT降低和NSF增多(P<0.05),鞘内注射后T0.5~3.0 h期间,BCP组的PWT和NSF均保持不变,而BCP+BQ123组的PWT先升高后降低,NSF先减少后增多,PWT和NSF均于1.5 h达到最高和最少,3.0 h恢复至注射前水平。胫骨骨质X线片显示,sham组和sham+BQ123组胫骨骨质密度均匀、骨皮质连续无缺失;BCP组注射后14 d出现大范围骨破坏、骨皮质缺损严重;BCP+BQ123组股骨远端见较小骨破坏病灶,部分皮质缺损。与sham组、sham+BQ123组比较,BCP组、BCP+BQ123组脊髓ET1、ETAR mRNA和蛋白及p-ERK1/2蛋白相对表达量均升高,且BCP+BQ123组低于BCP组,差异有统计学意义(P<0.05)。结论 BCP疼痛反应与脊髓ET-1及ETAR有关,鞘内注射ETAR拮抗药可有效减轻BCP大鼠的疼痛反应,可能通过抑制脊髓ERK通路发挥调控作用。
Abstract
Objective To observe the extent to which intrathecal injection of endothelin A receptor (ETAR) antagonist alleviates the pain of bone cancer pain (BCP) rats and the effect on extracellular regulatory protein kinase (ERK) pathway.Methods Sixty rats were selected for intrathecal catheterization and randomly divided into the sham operation group, sham operation + ETAR antagonist (sham+BQ123) group, bone cancer pain (BCP) group, and bone cancer pain + ETAR antagonist (BCP+BQ123) group. The BCP group and BCP+BQ123 group were inoculated with Walker256 cells in the bone marrow cavity of the distal femur to establish a BCP rat model. On the 14th day of modeling, 14 rats in the BCP+BQ123 group and sham + BQ123 group were injected with 7 μl of BQ123 intrathecally while 13 rats in the BCP group and 14 rats in the sham group were injected intrathecally with the same amount of normal saline. Immediately before intrathecal injection and 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 hours after injection, the pain behavior of rats in each group was evaluated in terms of the mechanical paw withdrawal threshold (PWT) and the number of spontaneous flinches (NSF). After the last assessment of pain behavior, the severity of bone destruction in each group was assessed via imaging. The expressions of endothelin 1 (ET1), ETAR, ERK1/2 and p-ERK1/2 were detected by RT-qPCR and Western blot.Results Compared with the sham group and sham + BQ123 group, PWT decreased while NSF increased before injection in the BCP group and BCP + BQ123 group (P<0.05). At T0.5-3.0 h after intrathecal injection, the PWT and NSF of the BCP group remained unchanged, while the PWT of the BCP + BQ123 group increased before it decreased, while the NSF decreased before it increased. Both the PWT and NSF reached the maximum and minimum at 1.5 hours, and returned to the pre-injection level at 3.0 hours. Tibia bone X-ray showed that tibia bone density was uniform in the sham group and sham + BQ123 group, and there was no loss of the bone cortex. BCP group suffered extensive bone destruction and severe bone cortical defects 14 days after injection. In the BCP + BQ123 group, there were small lesions of bone destruction in the distal femur, and some cortical defects. Compared with the sham group and sham + BQ123 group, the relative expressions of ET1, ETAR mRNA and protein and p-ERK1/2 protein in the spinal cord of the BCP group and BCP + BQ123 group were increased, but were lower in the BCP + BQ123 group than in the BCP group (P<0.05).Conclusions The pain response of BCP is related to spinal cord ET-1 and ETAR. Intrathecal injection of ETAR antagonist can effectively improve the pain response of BCP rats, possibly by inhibiting the ERK pathway of the spinal cord.
关键词
骨癌痛 /
内皮素A受体 /
疼痛 /
细胞外调节蛋白激酶
Key words
bone cancer pain /
endothelin A receptor /
pain /
extracellular regulatory protein kinase
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Kirshner J J, McDonald M C 3rd, Kruter F, et al. NOLAN: a randomized, phase 2 study to estimate the effect of prophylactic naproxen or loratadine vs no prophylactic treatment on bone pain in patients with early-stage breast cancer receiving chemotherapy and pegfilgrastim[J]. Support Care Cancer, 2018, 26(4):1323-1334.
[2] 范金艳, 刘丽雅, 马 洋. 康复外科护理对骨癌疼痛患者行微创手术的影响[J]. 中华肿瘤防治杂志, 2018, 25(2): 253-254.
[3] 刘 红, 蒋远静, 刘水霞. 癌痛规范化治疗提高中重度癌痛患者生活质量的临床及护理分析[J]. 第三军医大学学报, 2018, 40(10): 934-938.
[4] Tai L W, Pan Z, Sun L, et al. Suppression of Pax2 attenuates allodynia and hyperalgesia through ET-1-ETAR-NFAT5 signaling in a rat model of neuropathic pain[J]. Neuroscience, 2018, 384:139-151.
[5] Portenoy R K, Ahmed E. Cancer pain syndromes[J]. Hematol Oncol Clin North Am, 2018, 32(3):371-386.
[6] 李晓青, 俞卫锋, 孙玉明, 等. Walker 256乳腺癌细胞构建大鼠胫骨骨癌痛模型[J]. 中国肿瘤生物治疗杂志, 2008, 15(1): 41-45.
[7] Ibrahim M M, Patwardhan A, Gilbraith K B, et al. Long-lasting antinociceptive effects of green light in acute and chronic pain in rats[J]. Pain, 2017, 158(2): 347-360.
[8] Harton L R, Richardson J R, Armendariz A, et al. Dissociation of morphine analgesic effects in the sensory and affective components of formalin-induced spontaneous pain in male and female rats[J]. Brain Res, 2017, 1658: 36-41.
[9] 钟丽红, 陈文宇, 林伟龙, 等. 中西医综合治疗骨癌疼痛的临床观察[J]. 中华中医药学刊, 2019, 37(2): 409-413.
[10] Zhang H, Lund D M, Ciccone H A, et al. Peripherally restricted cannabinoid 1 receptor agonist as a novel analgesic in cancer-induced bone pain[J]. Pain, 2018, 159(9):1814-1823.
[11] Gomes L O, Chichorro J G, Araya E I, et al. Facial hyperalgesia due to direct action of endothelin-1 in the trigeminal ganglion of mice[J]. J Pharm Pharmacol, 2018, 70(7): 893-900.
[12] Pontes R B, Lisboa M R P, Pereira A F, et al. Involvement of endothelin receptors in peripheral sensory neuropathy induced by oxaliplatin in mice[J]. Neurotox Res, 2019, 36(4): 688-699.
[13] Tang Y, Peng H, Liao Q, et al. Study of breakthrough cancer pain in an animal model induced by endothelin-1[J]. Neurosci Lett, 2016, 617:108-115.
[14] Mantyh P W. Mechanisms that drive bone pain across the lifespan[J]. Br J Clin Pharmacol, 2019, 85(6):1103-1113.
[15] Lee S H, Kwon S C, Ok S H, et al. Levobupivacaine-induced vasoconstriction involves caldesmon phosphorylation mediated by tyrosine kinase-induced ERK phosphorylation[J]. Eur J Pharmacol, 2019, 842:167-176.
[16] Shenoy P A, Kuo A, Khan N, et al. The somatostatin receptor-4 agonist J-2156 alleviates mechanical hypersensitivity in a rat model of breast cancer induced bone pain[J]. Front Pharmacol, 2018, 9: 495.
[17] Nakamura A, Ono H, Ando A, et al. Suppression of the acute upregulation of phosphorylated-extracellular regulated kinase in ventral tegmental area by a μ-opioid receptor agonist is related to resistance to rewarding effects in a mouse model of bone cancer[J]. J Pharmacol Sci, 2017, 133(1):9-17.
基金
青海省基础研究计划项目(2018-0302-ZJC-0042)
PDF(939 KB)
