CAJ | 학술논문

目的通过观察骨癌痛小鼠痛行为学的变化,磷酸化环磷酸腺苷反应元件结合蛋白(phosphorylated cAMP response element binding protein,pCREB)和脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)在脊髓背角中表达的变化,探讨pCREB和BDNF在骨癌痛产生和维持中的作用. 方法 52只雄性C3H/HeJ小鼠,采用随机数字表法分为骨癌痛组(T组)和假手术组(S组),每组26例.T组小鼠右侧股骨骨髓腔内注射含2×105个纤维肉瘤细胞的20μl最小必须培养基(α-minimal essence medium,α-MEM),而S组注入不含肿瘤细胞的20μ1α-MEM,分别于接种肿瘤细胞前1d、接种后第4、7、10、14、21天测定自发抬足次数(spontaneous lifting times,SLTs)和机械缩足阈值(paw withdrawal mechanical threshold,PWMT).按照分组在相应时间点处死小鼠,用免疫印迹法测定小鼠L3～5脊髓背角pCREB和BDNF的蛋白表达水平. 结果与术前基础值和S组比较:接种后7、10、14、21d,T组小鼠SLTs[(4.43±0.91),(7.10±1.03),(11.27±1.35),(13.03±0.58)次]显著增加(P＜0.05);接种后10、14、21 d,T组小鼠PWMT[(0.63±0.20)、(0.32±0.12)、(0.24±0.12)g]明显下降(P＜0.05),脊髓背角pCREB[(3.78±0.58)、(3.92±0.46)、(4.92±0.37)]和BDNF[(2.13±0.31)、(2.88±0.15)、(2.58±0.41)]的表达显著增加(P＜0.05). 结论骨癌痛小鼠脊髓背角pCREB和BDNF表达增加,并且与痛行为学的改变具有相关性,提示其可能参与了骨癌痛的产生和维持.
목적 통과관찰골암통소서통행위학적변화,린산화배린산선감반응원건결합단백(phosphorylated cAMP response element binding protein,pCREB)화뇌원성신경영양인자(brain-derived neurotrophic factor,BDNF)재척수배각중표체적변화,탐토pCREB화BDNF재골암통산생화유지중적작용. 방법 52지웅성C3H/HeJ소서,채용수궤수자표법분위골암통조(T조)화가수술조(S조),매조26례.T조소서우측고골골수강내주사함2×105개섬유육류세포적20μl최소필수배양기(α-minimal essence medium,α-MEM),이S조주입불함종류세포적20μ1α-MEM,분별우접충종류세포전1d、접충후제4、7、10、14、21천측정자발태족차수(spontaneous lifting times,SLTs)화궤계축족역치(paw withdrawal mechanical threshold,PWMT).안조분조재상응시간점처사소서,용면역인적법측정소서L3～5척수배각pCREB화BDNF적단백표체수평. 결과 여술전기출치화S조비교:접충후7、10、14、21d,T조소서SLTs[(4.43±0.91),(7.10±1.03),(11.27±1.35),(13.03±0.58)차]현저증가(P＜0.05);접충후10、14、21 d,T조소서PWMT[(0.63±0.20)、(0.32±0.12)、(0.24±0.12)g]명현하강(P＜0.05),척수배각pCREB[(3.78±0.58)、(3.92±0.46)、(4.92±0.37)]화BDNF[(2.13±0.31)、(2.88±0.15)、(2.58±0.41)]적표체현저증가(P＜0.05). 결론 골암통소서척수배각pCREB화BDNF표체증가,병차여통행위학적개변구유상관성,제시기가능삼여료골암통적산생화유지.
Objective To investigate the role of phosphorylated cyclic-AMP response element binding protein (pCREB) and brain-derived neurotrophic factor (BDNF) in bone cancer pain,via observing the change of pain behavior and the expression of pCREB and BDNF in dorsal horn of spinal cord.Methods A total of 52 male mice (C3H/HeJ) were randomly divided into tumor group (group T,n=26)and sham group (group S,n=26).In group T,bone cancer pain was induced by implantation 20 μl of α-minimal essence medium (α-MEM) containing osteosarcoma cells into the intramedullary space of right femur.In group S,20 μl culture medium α-MEM containing no cancer cells was injected instead.Pain behavior of animals in both groups was assessed by the spontaneous lifting times (SLTs) and paw withdrawal mechanical threshold (PWMT) measured at 1 day before and at 4,7,10,14 d,and 21 d after inoculation.Another 3 mice were sacrificed at the corresponding time point and the L3-5 spinal cords were obtained for determining the expressions of pCREB and BDNF by western blotting.Results Compared with preoperative base level,the levels of SLTs in group S were significantly increased at 7,10,14,21 d after inoculation,the obvious increasing of SLTs [(4.43±0.91),(7.10±1.03),(11.27 ±1.35),(13.03 ±0.58),respectively] (P＜0.05),at 10,14,21 d after inoculation,the significant decreasing of PWMT[(0.63±0.20),(0.32±0.12),(0.24±0.12) g] were observed in group T (P＜0.05) and the expressions of pCREB[(3.78±0.58),(3.92±0.46),(4.92±0.37)] and BDNF [(2.13±0.31),(2.88±0.15),(2.58±0.41)] in group T were obviously increased (P＜0.05).Conclusions The expressions of pCREB and BDNF in dorsal horns of spinal cords were increased in a mouse model of bone cancer pain,accompanied by the pain behavior,suggesting spinal cord pCREB and BDNF may be involved in the development and maintenance of bone cancer pain in mice.