中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
32期
2531-2534
,共4页
王寒琪%许世杰%姚明%刘明娟%徐龙生%倪华栋%连庆泉
王寒琪%許世傑%姚明%劉明娟%徐龍生%倪華棟%連慶泉
왕한기%허세걸%요명%류명연%서룡생%예화동%련경천
骨肿瘤%疼痛%受体,嘌呤能P2Y12%白细胞介素1β%白细胞介素6
骨腫瘤%疼痛%受體,嘌呤能P2Y12%白細胞介素1β%白細胞介素6
골종류%동통%수체,표령능P2Y12%백세포개소1β%백세포개소6
Bone neoplasms%Pain%Receptor,purinergic P2Y12%Interleukin-1 beta%Interleukin-6
目的：观察鞘内注射P2Y12受体抑制剂MRS2395对骨癌痛大鼠痛阈及其脊髓炎症因子表达的影响，探讨P2 Y12受体与炎症因子在骨癌痛发病机制中的可能作用。方法雌性SD大鼠32只，随机数字表法随机分为4组：假手术组（ S组）、MRS2395组（ M组）、胫骨癌痛组（ A组）、胫骨癌痛MRS2395组（ MA组）。 S组、M组左胫骨上端骨髓腔各注入Hank′s液10μl，A组、MA组左胫骨上端骨髓腔各注入Walker256癌细胞悬液10μl。骨癌痛造模同时均进行L3-4间隙鞘内置管，模型制备成功后第9～12天， S 组、A 组鞘内注射0.9％生理盐水15μl/d， M 组、MA 组鞘内注射 MRS2395（400 pmol/μl）15μl/d。术后第9～12天鞘内注药前、后每10分钟测试1次大鼠左后足机械痛阈值，术后第12天鞘内注药测定机械痛阈值后，取L4-L6左侧脊髓背角，采用酶联免疫吸附测定法测脊髓炎症因子IL-1β、IL-6的表达情况。结果建模后第9天，S组、M组、A组、MA组给药后20 min机械痛阈值分别为（34.2±5.8）g、（34.4±5.7）g、（21.0±2.0）g、（25.4±2.3）g，差异有统计学意义（F＝18.679，P＜0.01）；与S组、M组比较，A组机械痛阈值下降；与A组比较，MA组机械痛阈值增高。建模后第12天，S组、M组、A组、MA组大鼠脊髓IL-1β含量分别为（74.0±18.6）pg/ml、（98.4±17.3） pg/ml、（253.5±66.4）pg/ml、（146.3±22.3） pg/ml，差异有统计学意义（F＝18.221，P＜0.01）；与S组、M组比较，A组大鼠脊髓IL-1β含量明显升高；S组、M组、A组、MA组大鼠脊髓IL-6含量分别为（377.4±65.8）pg/ml、（331.6±67.9） pg/ml、（856.1±53.4） pg/ml、（596.1±34.9） pg/ml，差异有统计学意义（F＝70.880，P＜0.01）；与S组、M组比较，A组大鼠脊髓IL-6含量明显升高；与A组比较， MA组大鼠鞘内注射MRS2395后IL-1β和IL-6含量均明显降低。结论鞘内注射MRS2395可缓解骨癌痛大鼠的痛觉过敏，抑制骨癌痛大鼠脊髓内炎症因子表达上调，P2Y12受体可能通过调节IL-1β、IL-6的表达参与骨癌痛的发生。
目的：觀察鞘內註射P2Y12受體抑製劑MRS2395對骨癌痛大鼠痛閾及其脊髓炎癥因子錶達的影響，探討P2 Y12受體與炎癥因子在骨癌痛髮病機製中的可能作用。方法雌性SD大鼠32隻，隨機數字錶法隨機分為4組：假手術組（ S組）、MRS2395組（ M組）、脛骨癌痛組（ A組）、脛骨癌痛MRS2395組（ MA組）。 S組、M組左脛骨上耑骨髓腔各註入Hank′s液10μl，A組、MA組左脛骨上耑骨髓腔各註入Walker256癌細胞懸液10μl。骨癌痛造模同時均進行L3-4間隙鞘內置管，模型製備成功後第9～12天， S 組、A 組鞘內註射0.9％生理鹽水15μl/d， M 組、MA 組鞘內註射 MRS2395（400 pmol/μl）15μl/d。術後第9～12天鞘內註藥前、後每10分鐘測試1次大鼠左後足機械痛閾值，術後第12天鞘內註藥測定機械痛閾值後，取L4-L6左側脊髓揹角，採用酶聯免疫吸附測定法測脊髓炎癥因子IL-1β、IL-6的錶達情況。結果建模後第9天，S組、M組、A組、MA組給藥後20 min機械痛閾值分彆為（34.2±5.8）g、（34.4±5.7）g、（21.0±2.0）g、（25.4±2.3）g，差異有統計學意義（F＝18.679，P＜0.01）；與S組、M組比較，A組機械痛閾值下降；與A組比較，MA組機械痛閾值增高。建模後第12天，S組、M組、A組、MA組大鼠脊髓IL-1β含量分彆為（74.0±18.6）pg/ml、（98.4±17.3） pg/ml、（253.5±66.4）pg/ml、（146.3±22.3） pg/ml，差異有統計學意義（F＝18.221，P＜0.01）；與S組、M組比較，A組大鼠脊髓IL-1β含量明顯升高；S組、M組、A組、MA組大鼠脊髓IL-6含量分彆為（377.4±65.8）pg/ml、（331.6±67.9） pg/ml、（856.1±53.4） pg/ml、（596.1±34.9） pg/ml，差異有統計學意義（F＝70.880，P＜0.01）；與S組、M組比較，A組大鼠脊髓IL-6含量明顯升高；與A組比較， MA組大鼠鞘內註射MRS2395後IL-1β和IL-6含量均明顯降低。結論鞘內註射MRS2395可緩解骨癌痛大鼠的痛覺過敏，抑製骨癌痛大鼠脊髓內炎癥因子錶達上調，P2Y12受體可能通過調節IL-1β、IL-6的錶達參與骨癌痛的髮生。
목적：관찰초내주사P2Y12수체억제제MRS2395대골암통대서통역급기척수염증인자표체적영향，탐토P2 Y12수체여염증인자재골암통발병궤제중적가능작용。방법자성SD대서32지，수궤수자표법수궤분위4조：가수술조（ S조）、MRS2395조（ M조）、경골암통조（ A조）、경골암통MRS2395조（ MA조）。 S조、M조좌경골상단골수강각주입Hank′s액10μl，A조、MA조좌경골상단골수강각주입Walker256암세포현액10μl。골암통조모동시균진행L3-4간극초내치관，모형제비성공후제9～12천， S 조、A 조초내주사0.9％생리염수15μl/d， M 조、MA 조초내주사 MRS2395（400 pmol/μl）15μl/d。술후제9～12천초내주약전、후매10분종측시1차대서좌후족궤계통역치，술후제12천초내주약측정궤계통역치후，취L4-L6좌측척수배각，채용매련면역흡부측정법측척수염증인자IL-1β、IL-6적표체정황。결과건모후제9천，S조、M조、A조、MA조급약후20 min궤계통역치분별위（34.2±5.8）g、（34.4±5.7）g、（21.0±2.0）g、（25.4±2.3）g，차이유통계학의의（F＝18.679，P＜0.01）；여S조、M조비교，A조궤계통역치하강；여A조비교，MA조궤계통역치증고。건모후제12천，S조、M조、A조、MA조대서척수IL-1β함량분별위（74.0±18.6）pg/ml、（98.4±17.3） pg/ml、（253.5±66.4）pg/ml、（146.3±22.3） pg/ml，차이유통계학의의（F＝18.221，P＜0.01）；여S조、M조비교，A조대서척수IL-1β함량명현승고；S조、M조、A조、MA조대서척수IL-6함량분별위（377.4±65.8）pg/ml、（331.6±67.9） pg/ml、（856.1±53.4） pg/ml、（596.1±34.9） pg/ml，차이유통계학의의（F＝70.880，P＜0.01）；여S조、M조비교，A조대서척수IL-6함량명현승고；여A조비교， MA조대서초내주사MRS2395후IL-1β화IL-6함량균명현강저。결론초내주사MRS2395가완해골암통대서적통각과민，억제골암통대서척수내염증인자표체상조，P2Y12수체가능통과조절IL-1β、IL-6적표체삼여골암통적발생。
Objective To explore the roles of P2Y12 receptor ( P2Y12R) in bone cancer pain by observing the changes of inflammatory cytokines (IL-1β, IL-6) after intrathecal injection (i.t.) of P2Y12R antagonist MRS2395.Methods Thirty-two female SD rats were randomly divided into 4 groups ( n =8 each):sham group (group S), MRS2395 group (group M), cancer group (group A) and cancer +MRS2395 group ( group MA).Groups S and M received an injection of 10 μl Hank′s solution into left tibia medullar cavity while groups A and MA had an injection of Walker 256 mammary cancer cells (10 μl, 2 × 107 cells/ml) into the same place.At Day 9-12 post-inoculation, groups S and A received an injection of saline (0.9%, 15 μl, i.t.) while groups M and MA had MRS2395 (400 pmol/μl, 15 μl, i.t.).Intrathecal catheterization between L 3 and L4 was performed immediately after inoculating tumor cells by inserting a small tube into vertebral space.Mechanical withdrawal thresholds were measured on left hind paws before and during 10-minute intervals after dosing.Spinal cords ( L4-L6 segments ) were removed for determining the expressions of IL-1βand IL-6 by enzyme-linked immunosorbent assay ( ELISA ) at Day 12 after drug delivery.Results At 20 min post-injection, mechanical withdrawal thresholds of groups S , M, A and MA were (34.2 ±5.8), (34.4 ±5.7), (21.0 ±2.0) and (25.4 ±2.3) g respectively at Day 9 post-inoculation ( F=18.679, P <0.01 ); mechanical withdrawal thresholds of group A obviously decreased versus groups S and M;mechanical withdrawal thresholds in group MA increased obviously versus group A.The expressions of IL-1βin groups S , M, A and MA were ( 74.0 ±18.6 ) , ( 98.4 ±17.3 ) , ( 253.5 ± 66.4) and (146.3 ±22.3) pg/ml at Day 12 post-inoculation (F=18.221, P<0.01); compared with groups S and M, the expression of IL-1βin group A showed a significant up-regulation.Likewise, the expressions of IL-6 were (377.4 ±65.8), (331.6 ±67.9), (856.1 ±53.4) and (596.1 ±34.9) pg/ml (F=70.880, P<0.01) respectively in groups S, M, A and MA; compared with groups S and M, the expression of IL-6 increased obviously in group A.There were significant decreases of IL-1βand IL-6 in group MA versus group A .Conclusions An intrathecal injection of MRS 2395 may alleviate hyperalgesia and inhibit the up-regulated expression of spinal cord inflammatory cytokines in bone cancer rats.And P2Y12 receptor may be involved in the formation of bone cancer pain through regulating the expressions of IL-1βand IL-6.
