CAJ | 학술논문

目的评价脊髓小胶质细胞CC趋化因子受体2(CCR2)在大鼠骨癌痛维持中的作用.方法健康雌性未交配SD大鼠50只,2月龄,体重160～ 180 g,采用随机数字表法,将其分为5组(n=10):假手术组(Ⅰ组)、假手术+RS102895(CCR2拮抗剂)组(Ⅱ组)、骨癌痛组(Ⅲ组)、骨癌痛+DMSO组(Ⅳ组)、骨癌痛+RS102895组(Ⅴ组).左侧胫骨干骺端骨髓腔内接种Walker256乳腺癌细胞制备大鼠骨癌痛模型.于术后10-12 d,Ⅱ组和Ⅴ组分别鞘内注射3 μg/μl RS102895 10μl,Ⅳ组鞘内注射DMSO 10μl,Ⅰ组和Ⅲ组鞘内注射生理盐水10μl,1次/d.于术前1d、术后3、6、9、10、11、12 d时测定机械痛阈,术后12 d痛阈测定后,取L4-6脊髓组织,采用免疫组织化学法测定脊髓背角小胶质细胞激活标记物(ox-42)水平,采用ELISA法测定脊髓IL-1β、IL-6及TNF-α的含量.结果与Ⅰ组比较,Ⅲ组、Ⅳ组、Ⅴ组术后6-12 d时机械痛阈降低,ox-42阳性细胞数、脊髓IL-1β、IL-6及TNF-α含量升高(P＜0.01),Ⅱ组上述指标差异无统计学意义(P＞0.05);与Ⅲ组比较,Ⅴ组术后10-12 d时机械痛阈升高,ox-42阳性细胞数、脊髓IL-1β、IL-6及TNF-α含量降低(P＜0.01),Ⅳ组上述指标差异无统计学意义(P＞0.05).结论脊髓小胶质细胞CCR2参与大鼠骨癌痛的维持,其机制可能与激活小胶质细胞从而促进炎性细胞因子释放有关.
목적 평개척수소효질세포CC추화인자수체2(CCR2)재대서골암통유지중적작용.방법 건강자성미교배SD대서50지,2월령,체중160～ 180 g,채용수궤수자표법,장기분위5조(n=10):가수술조(Ⅰ조)、가수술+RS102895(CCR2길항제)조(Ⅱ조)、골암통조(Ⅲ조)、골암통+DMSO조(Ⅳ조)、골암통+RS102895조(Ⅴ조).좌측경골간후단골수강내접충Walker256유선암세포제비대서골암통모형.우술후10-12 d,Ⅱ조화Ⅴ조분별초내주사3 μg/μl RS102895 10μl,Ⅳ조초내주사DMSO 10μl,Ⅰ조화Ⅲ조초내주사생리염수10μl,1차/d.우술전1d、술후3、6、9、10、11、12 d시측정궤계통역,술후12 d통역측정후,취L4-6척수조직,채용면역조직화학법측정척수배각소효질세포격활표기물(ox-42)수평,채용ELISA법측정척수IL-1β、IL-6급TNF-α적함량.결과 여Ⅰ조비교,Ⅲ조、Ⅳ조、Ⅴ조술후6-12 d시궤계통역강저,ox-42양성세포수、척수IL-1β、IL-6급TNF-α함량승고(P＜0.01),Ⅱ조상술지표차이무통계학의의(P＞0.05);여Ⅲ조비교,Ⅴ조술후10-12 d시궤계통역승고,ox-42양성세포수、척수IL-1β、IL-6급TNF-α함량강저(P＜0.01),Ⅳ조상술지표차이무통계학의의(P＞0.05).결론 척수소효질세포CCR2삼여대서골암통적유지,기궤제가능여격활소효질세포종이촉진염성세포인자석방유관.
Objective To evaluate the role of spinal microglial C-C chemokine receptor type 2 (CCR2) in the maintenance of bone cancer pain (BCP) in rats.Methods Fifty unmated female Sprague-Dawley rats,aged 2 months,weighing 160-180 g,were randomly divided into 5 groups (n =10 each):sham operation group (group Ⅰ),sham operation + RS102895 (CCR2 antagonist) group (group Ⅱ),BCP group (group Ⅲ),BCP + dimethylsulfoxide (DMSO) group (group Ⅳ),and BCP + RS102895 group (group Ⅴ).The rats were anesthetized with intraperitoneal chloral hydrate.BCP was induced by intra-tibial inoculation of 1 × 105 Walker 256 mammary gland carcinoma cells into the medullary cavity of the left tibial metaphysis.On 10-12 days after operation,3 μg/μl RS102895 10 μd was injected intrathecally once a day in Ⅱ and Ⅴ groups,10％ DMSO 10 μl was injected intrathecally once a day in Ⅳ group,and normal saline 10 μl was injected intrathecally once a day in Ⅰ and Ⅲ groups.Mechanical paw withdrawal threshold was measured at 1 day before operation and 3,6,9,10,11 and 12 days after operation.After measurement of pain threshold at day 12 after operation,the lumbar segments (L4-6) of the spinal cord were removed for determination of the level of ox-42 (spinal microglial activation marker) (by immuno-histochemistry) and contents of IL1-β,IL-6 and TNF-α (by ELISA).Results Compared with group Ⅰ,mechanical paw withdrawal threshold was significantly decreased at 6-12 days after operation,the number of ox-42 positive cells and contents of IL1-β,IL-6 and TNF-α were increased in Ⅲ,Ⅳ and Ⅴ groups,and no significant change was found in the parameters mentioned above in group Ⅱ.Compared with group Ⅲ,mechanical paw withdrawal threshold was significantly increased at 10-12 days after operation,the number of ox-42 positive cells and contents of IL1-β,IL-6 and TNF-α were decreased in group Ⅴ,and no significant change was found in the parameters mentioned above in group Ⅳ.Conclusion Spinal microglial CCR2 is involved in the maintenance of BCP via activating microglia and promoting the release of inflammatory cytokines of rats.