摘要
骨癌痛(BCP)是恶性肿瘤患者最常见的疼痛之一,严重影响患者的生活质量。BCP的分子作用机制和新药研发都迫在眉睫。2-溴棕榈酸(2-BP)作为一种蛋白质棕榈化抑制剂在病理性疼痛中有镇痛效果,而在骨癌痛中作用仍不清楚。酸敏感离子通道3型(ASIC3),作为一个重要的疼痛因子能否受到2-BP的调控也未知。为了检测2-BP在骨癌痛中的作用,并研究其对背根神经节(DRG)中ASIC3的调控,本文开展了相关工作。1)首先建立BCP大鼠模型,将大鼠乳腺癌细胞(MRMT-1)注射入雌大鼠胫骨骨髓腔内,21 d后通过X射线和机械痛检测,发现与假性手术组相比,BCP模型大鼠的胫骨被破坏;同时,BCP组大鼠的机械疼痛值明显上升(假性手术组PWT vs.BCP PWT:16.1±1.5 vs.5.3±1.5;P<0.01);表明大鼠乳腺癌骨转移疼痛模型成功构建。2)蛋白质免疫印迹检测结果显示,与正常和假性手术组相比,BCP大鼠L4-L6 DRG中酸敏感离子通道3蛋白表达上调(0.63±0.03,0.64±0.1和1.07±0.05)。3)在术后第21 d,给BCP大鼠腹腔注射2-BP,发现给药组BCP大鼠的机械疼痛值下调(6 h后,PWT对照vs.PWT 2-BP:6.9±2.0 vs.10.8±1.6,P<0.01),表明2-BP在骨癌痛模型大鼠中具有镇痛作用。4)蛋白质免疫印迹结果显示,与给药前相比,2-BP处理后降低了BCP大鼠L4-L6 DRG中膜上ASIC3蛋白的表达(1.05±0.13,0.66±0.12)。同时,在ASIC3介导的酸痛模型中,2-BP给药降低大鼠震颤的次数(对照组为27±1.8次,2-BP组为10±1.5次),表明2-BP给药阻断ASIC3介导的酸痛。5)在ASIC3转染的SH-SY5Y细胞中,与对照相比,2-BP给药后明显降低膜上ASIC3蛋白表达量(1.0±0.2,0.58±0.10)。这些结果表明,2-BP在骨癌痛中具有镇痛作用,其镇痛机制涉及到调控背根神经节中膜上酸敏感离子通道3的表达。
Bone cancer pain（BCP） is one of the most common pains in patients with malignant cancers,and greatly affects the quality of patient life. Understanding the mechanism and improving novel agents for BCP treatment is desirable. As an inhibitor of protein palmitoylation,2-bromopalmitate（2-BP）improved pain behaviors in neuropathic pain model,whereas its function on BCP was unclear. Acidsensing ion channel 3（ASIC3） plays an important role in various pain conditions. However,whether ASIC3 expression was regulated by 2-BP was unknown. In order to investigate the effect of 2-BP on BCP and detect the regulation of 2-BP on ASIC3,we did these research. First,we established BCP rats model through intrathecally injecting rat mammary gland carcinoma cells（MRMT-1） into the left tibia of Sprague-Dawley female rats. On post-operative day（POD） 21,tibia radiographs of BCP rats showed regional bone destruction and damage. And mechanical pain sensitivities of BCP rats were significantly increased compared with sham rats（Sham PWT vs BCP PWT： 16. 1 ± 1. 5 vs 5. 3 ± 1. 5; P 0. 01）,indicating that BCP rats model was working. Second,compared with normal and sham rats,ASIC3 expression in L4-L6 DRG neurons of BCP rats was significantly increased（0. 63 ± 0. 03,0. 64 ± 0. 1 and 1. 07 ± 0. 05）. Third,compared to the vehicle-treated BCP rats,2-BP treated BCP rats resulted in a significant increase in PWT（at post injection 6 h,PWT vehicle vs PWT 2-BP： 6. 9 ± 2. 0 vs 10. 8 ±1. 6,P 〈0. 01）,indicating 2-BP treatment had an antinociceptive effect in BCP rats. Forth,after 2-BP treated,membrane ACIS3（M-ASIC3） expression in DRG of BCP rats decreased compared with that before 2-BP treated（1. 05 ± 0. 13,0. 66 ± 0. 12）. And in ASIC3-mediated acidosis pain model,2-BP treatment decreased the number of flinches（vehicle treated rats： 27 ± 1. 8,2-BP treated rats： 10 ±1. 5）,indicating 2-BP treatment blocked ASIC3-mediated acidosis pain. Fifth,the levels of M-ASIC3 were reduced after treatment with 2-BP in ASIC3-transfected SH-SY5 Y cells（1. 0 ± 0. 2,0. 58 ±0. 10）. These data illustrate that 2-BP attenuated bone cancer pain by reducing membrane ASIC3 expression levels in DRG neurons.
作者
唐琼
李鸣粤
余良主
朱海丽
TANG Qiong;LI Ming-Yue;Yu Liang-Zhu;ZHU Hai-Li(Research Center of Basic Medical Sciences,Department of Physiology,School of Basic Medical Sciences Hubei University of Science and Technology,Xianning 437100,Hubei,China;）School of Pharmacy,Department of Pharmaceutical Preparations,Hubei University of Science and Technology,Xianning 437100,Hubei,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2018年第8期884-890,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
Supported by National Natural Science Foundation of China(No.81641154)
Research Project of Hubei Provincial Department of Education(No.Q20172804)~~
关键词
骨癌痛
2-溴棕榈酸
背根神经节
酸敏感离子通道3型
bone cancer pain（BCP）
2-bromopalmitate（BP）
dorsal root ganglion （DRG）
acid-sensingion channels 3 （ASIC3）
