CAJ | 학술논문

目的评价CX3C趋化因子受体1(CX3 CR1)对骨癌痛大鼠吗啡耐受时脊髓背角μ受体和辣椒素受体(TRPVl)表达的影响.方法清洁级成年雌性SD大鼠,体重180-200 g,月龄3月,经L3.4棘突间隙行鞘内置管,取鞘内置管成功的大鼠48只,采用随机数字表法,将大鼠随机分为4组(n=12):对照组(A组)、鞘内注射生理盐水组(AM组)、鞘内注射IgG组(GM组)和鞘内注射CX3CR1中和抗体组(BM组).A组仪手术暴露右侧胫骨七段；其余3组右侧胫骨上段骨髓腔注入Walker256 乳腺癌细胞10μl(400个/ μl)建立骨癌痛模型,术后第10天开始鞘内注射吗啡20tg/kg,2次/d,连续7d,建立骨癌痛-吗啡耐受模型,注射吗啡第8天分别经鞘内注射相应溶液10μl,1次/d,连续3d.分别于接种Walker 256乳腺癌细胞前(T0)、接种后第3、6、9天、注射吗啡第3、7天、鞘内注射抗体第3天(T1-T6)时测定机械缩足阈值(MWT)和机械缩足持续时间(MWD),于T6时测定痛阈后处死大鼠,取脊髓L4-6节段组织,采用Western blot法检测脊髓背角小胶质细胞CX3 CR1蛋白的表达,采用免疫组化法检测神经元μ受体和TRPV1的表达.结果与A组比较,BM组T2.3.5时、AM组和GM组T2,3,5,6时MWT下降,MWD升高,T6时CX3CR1蛋白和TRPV1表达上调,μ受体表达下调(P＜0.01)；与AM组和GM组比较,BM组T6时MWT上升,MWD降低,CX3CR1蛋白和TRPV1表达下调,μ受体表达上调(P＜0.01).AM组和GM组上述指标差异无统计学意义(p＞0.05).结论 CX3CR1可通过下调大鼠脊髓背角μ受体和上调TRPVI参与骨癌痛大鼠吗啡耐受的形成.
목적 평개CX3C추화인자수체1(CX3 CR1)대골암통대서마배내수시척수배각μ수체화랄초소수체(TRPVl)표체적영향.방법 청길급성년자성SD대서,체중180-200 g,월령3월,경L3.4극돌간극행초내치관,취초내치관성공적대서48지,채용수궤수자표법,장대서수궤분위4조(n=12):대조조(A조)、초내주사생리염수조(AM조)、초내주사IgG조(GM조)화초내주사CX3CR1중화항체조(BM조).A조의수술폭로우측경골칠단；기여3조우측경골상단골수강주입Walker256 유선암세포10μl(400개/ μl)건립골암통모형,술후제10천개시초내주사마배20tg/kg,2차/d,련속7d,건립골암통-마배내수모형,주사마배제8천분별경초내주사상응용액10μl,1차/d,련속3d.분별우접충Walker 256유선암세포전(T0)、접충후제3、6、9천、주사마배제3、7천、초내주사항체제3천(T1-T6)시측정궤계축족역치(MWT)화궤계축족지속시간(MWD),우T6시측정통역후처사대서,취척수L4-6절단조직,채용Western blot법검측척수배각소효질세포CX3 CR1단백적표체,채용면역조화법검측신경원μ수체화TRPV1적표체.결과 여A조비교,BM조T2.3.5시、AM조화GM조T2,3,5,6시MWT하강,MWD승고,T6시CX3CR1단백화TRPV1표체상조,μ수체표체하조(P＜0.01)；여AM조화GM조비교,BM조T6시MWT상승,MWD강저,CX3CR1단백화TRPV1표체하조,μ수체표체상조(P＜0.01).AM조화GM조상술지표차이무통계학의의(p＞0.05).결론 CX3CR1가통과하조대서척수배각μ수체화상조TRPVI삼여골암통대서마배내수적형성.
Objective To investigate the effects of intrathecal(IT)CX3 CR1 neutralizing antibody(antiFKR)on morphine tolerance in rats with bone cancer pain(BCP)and the unlerlying mechanism.Methods Forty-eight adult female SD rats aged 3 months weighing 180-200 g were randomized into 4 groups(n =12 each):group I sham operation(S); group Ⅱ BCP + normal saline(NS); group Ⅲ BCP + IgG(IgG)and group ⅣBCP + anti-FKR.Bone cancer pain(BCP)was induced by injecting Walker 256 cancer cells 10 μl(400 cells/ μl)into the medullary cavity of right tibia in groups Ⅱ,Ⅲ and Ⅳ.Ten days later morphine 20 μg/kg was administered IT twice a day for 7 consecutive days.Starting from the 8th day NS,IgG and anti-KFR 10 μl was administered IT once a day for 3 consecutive days in groups Ⅱ,Ⅲ and ⅣⅣ respectively.Paw withdrawal threshold to yon Frey filament stimulation(MWT)and paw withdrawal duration(MWD)were determined bcfore(To,baseline)and at 3,6,9 day after intra-tibial cancer cell inoculation(T1.2,3),on the 3rd and 7th day of IT morphine(T4.5)and on the 3rd day of IT NS/lgG/anti-KFR(T6).The animals were killed at T6 after last pain behavior assessment.The lumbar segment(L4-6)of the spinal cord was removed for determination of the expression of CX3 CR1 protein(by Western blot),μ-opioid receptor and TRPV1 receptor(by immuno-histochemistry)in the dorsal horn of spinal cord.Results IT morphine significantly eased BCP at T4,but morphine analgesia was significantly reduced on the 7th day of IT morphine in the 3 groups indicating morphine tolerance which was significantly relieved by anti-KFR in group Ⅳ.IT anti-KFR significantly down-regulated CX3CR1 prolein and TRPVI receptor expression and up-regulated μ-opioid receptor in group Ⅳ as compared with IT NS and lgG in groups Ⅱ and Ⅲ.Conctusion IT anti-KFR can relieve morphine tolerance in the rats with bone cancer pain by up-regulating μ-opioid receptor and down-regulating CX3 CR1 protein and TRPVI receptor expression.