NLRP3炎性小体和IL-1β在骨癌痛大鼠吗啡耐受形成中的作用

The Role of NLRP3 Inflammatory Body and IL-1β in Morphine Tolerance of Rats With Bone Cancer Pain

目的评价NLRP3炎性小体和IL—1β在骨癌痛大鼠吗啡耐受形成中的作用及鞘内注射Anti-NLRP3对大鼠痛觉敏化的的影响。方法将鞘内置管成功的大鼠随机分为5组,每组各10例,假手术组(S组)、骨癌痛组(P组)、骨癌痛+吗啡耐受组(PM组)、骨癌痛+吗啡耐受+Ig G组(PMG组)、骨癌痛+吗啡耐受+Anti-NLRP3组(PMA组)。除S组外大鼠接种Walker 256乳腺癌细胞制备骨癌痛模型,接种后的第14 d起,给PM组、PMG组和PMA组大鼠连续7 d皮下注射吗啡,构建吗啡耐受模型。第21 d时,五组大鼠注射吗啡3 mg/kg。造模后第22 d起,连续5 d给PM组大鼠鞘内注射生理盐水10μl,PMG组大鼠注射Ig G 10μl,PMA注射Anti-NLRP3 10μl。选取不同的时间点对大鼠行为学进行测试,所有测定完成后处死大鼠,取脊髓组织,测定NLRP3蛋白和IL-1β的表达。结果与S组和P组比较,PM、PMG和PMA组大鼠产生吗啡耐受后脊髓NLRP3蛋白和IL-1β表达上调(P<0.05)。鞘内注射Anti-NLRP3的PMA组与PM、PMG组相比,大鼠行走痛评分下降,PWMT升高,脊髓NLRP3蛋白和IL-1β表达下调(P<0.05)。结论 NLRP3炎性小体和IL-1β与大鼠骨癌痛-吗啡耐受形成有密切关系。鞘内注射Anti-NLRP3可减轻骨大鼠的痛觉敏化。

Objective To evaluate the role of NLRP3 inflammasome and il-1β on morphine tolerance in rats with bone cancer pain and the effects of anti-NLRP3 intrathecal injection on the hyperalgesia of rats.Methods Rats with successful intrathecal catheters were randomly divided to five groups with 10 rats in each group：sham operation group（S group）,bone cancer pain group（P group）,BCP ＋ morphine tolerance group（PM group）,BCP ＋ morphine tolerance group ＋ Ig G group（PM Ggroup）,BCP ＋ morphine tolerance group ＋ Anti-NLRP3 group（PMA group）.Walker 256 breast cancer cells were inoculated on rats except for rats in S group to establish the bone cancer pain model.On the 14 th day after inoculation,morphine was injected subcutaneously into rats in the PM group,PMG group and PMA group for 7 days to establish the morphine tolerance model.On the 21 st day,rats in the five groups were injected with 3 mg/kg morphine.22 days after modeling,10μl of saline was injected intrathecally into rats in the PM group for 5 consecutive days.10 μl of Ig G was injected into rats in the PMG group,and 10μl of anti-NLRP3 was injected into rats in the PMA group.Behaviors were tested at different time.Rats were sacrificed on the last day after all the experiments were completed,spinal cord segments were extracted,and the expression of NLRP3 and IL-1β was detected.Results Compared with rats in S and gruop P,expressions of NLRP3 proteins and IL-1βin segments of rats in PM、PMG and PMA group were increased（P〈0.05）.The free exercise pain score was decreased,PWMT was increased;expressions of NLRP3 proteins and IL-1βin segments of rats were decreased of rats in PMA group compared with those in PM and PMG group（P〈0.05）.Conclusion NLRP3 inflammasome and IL-1βare closely related to morphine tolerance of BCP in rats.Intrathecal injection of Anti-NLRP3 can alleviate hyperalgesia in morphinetolerant rats with BCP.