柯里拉京对吗啡耐受大鼠脊髓小胶质细胞活化的作用机制

Study on the Effects of Corilagin on Microglia Activation in the Spinal Cord of Morphine-tolerant Rats

目的 探讨柯里拉京对吗啡耐受大鼠的影响及作用机制。方法 建立慢性吗啡镇痛大鼠耐受模型。采用水浴甩尾法检测大鼠甩尾潜伏期,利用免疫荧光检测离子化钙结合适配分子1(Iba-1)的阳性细胞率;蛋白免疫印迹法检测Iba-1和丝裂原活化蛋白激酶(MAPK)信号通路相关蛋白的表达;采用酶联免疫吸附实验(ELISA)检测白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)表达;采用逆转录-聚合酶链反应(qRT-PCR)检测IL-6、TNF-α、IL-1β的mRNA表达。结果 与吗啡组相比,吗啡+柯里拉京组镇痛效果随柯里拉京浓度升高而增加。与对照组相比,吗啡组中Iba-1的阳性细胞率、Iba-1蛋白表达水平、炎性因子IL-6,TNF-α,IL-1β表达水平以及MAPK信号通路相关蛋白细胞外调节蛋白激酶(ERK)、c-Jun氨基末端激酶(JNK)和丝裂原活化蛋白激酶p38抗体(p38)的磷酸化水平显著增加,但在吗啡+柯里拉京组中随柯里拉京浓度升高而下降。吗啡和(或)柯里拉京处理对ERK、JNK和p38表达水平没有显著影响。结论 柯里拉京通过抑制MAPK通路降低大鼠吗啡耐受。

Objective This study was designed to investigate the effects of corilagin on morphine-tolerant rats and discuss the reaction mechanism.Methods A rat model of chronic morphine analgesia tolerance was established.The tail-flick latency of rats was detected by using the water bath tail-flick method.The positive cell rate of ionized calcium binding adapter molecule-1(Iba-1)was detected by using immunofluorescence assay.The protein expressions of Iba-1and mitogen-activated protein kinase(MAPK)pathway-related proteins were detected by Western blotting.The release of interleukin-6(IL-6),tumor necrosis factor alpha(TNF-α)and interleukin-1beta(IL-1β)was detected by using enzyme-linked immunosorbent assay(ELISA).The mRNA levels of IL-6,TNF-αand IL-1βwere detected by using quantitative reverse transcriptase polymerase chain reaction(qRT-PCR).Results Compared with the morphine(Mor)group,the analgesic effect of morphine in morphine+corilagin(Mor+Cor)group was strengthened with the increase of corilagin concentration.Compared with the Control group,the positive cell rate of Iba-1,Iba-1protein expression level,levels of inflammatory factors IL-6,TNF-α,IL-1β,and the phosphorylated expression levels of MAPK signaling pathway-related proteins extracellular regulated protein kinases(ERK),c-Jun N-terminal kinase(JNK)and p38 were significantly increased in the Mor group,which were then decreased in Mor+Cor group with the increase of corilagin concentration.Morphine and corilagin treatment(alone or in combination)had no significant effects on the expression levels of ERK,JNK and p38.Conclusion Corilagin reduces morphine tolerance in rats by inhibiting MAPK pathway.