氯吡格雷对疼痛-抑郁共病小鼠海马组织炎性因子及小胶质细胞的影响

Effect of clopidogrel on inflammatory cytokines and microglial in mice with comorbidity of pain and depression

目的 研究氯吡格雷对疼痛-抑郁共病模型小鼠行为学、海马组织炎性因子及小胶质细胞的影响。方法 选择SPF级雄性C57BL/6J小鼠24只,8周龄,体重23~27 g。采用随机数字表法将小鼠分为三组:假手术组(C组)、保留坐骨神经损伤(SNI)组(S组)和SNI+氯吡格雷组(L组),每组8只。C组手术过程与制备SNI模型一致,但不损伤神经,保持神经完整;S组制备SNI模型;L组在制备SNI模型后21 d起给予氯吡格雷10 mg/kg灌胃,连续14 d。于术前1 d、术后7、14、21、28、35 d检测机械缩足反射阈值(MWT)。于术后35、37 d分别进行悬尾实验(TST)和强迫游泳实验(FST),并计算不动时间。行为学实验后处死小鼠,取右侧海马组织,采用ELISA法检测IL-1β、IL-6、TNF-α浓度,采用RT-PCR法检测IL-1β、IL-6、TNF-α mRNA表达量,采用免疫荧光染色法检测小胶质细胞数量。结果 与C组比较,S组和L组术后7、14、21、28、35 d MWT明显降低,TST和FST不动时间明显延长,海马组织IL-1β、IL-6和TNF-α浓度和mRNA表达量明显升高,小胶质细胞数量明显增多(P<0.05)。与S组比较,L组术后28、35 d MWT明显升高,TST和FST不动时间明显缩短,海马组织IL-1β、IL-6和TNF-α浓度和mRNA表达量明显降低,小胶质细胞数量明显减少(P<0.05)。结论氯吡格雷可以降低疼痛-抑郁共病小鼠海马组织IL-1β、IL-6和TNF-α浓度和mRNA表达量,减轻炎症反应,减少小胶质细胞数量,改善慢性神经病理性痛和抑郁。

Objective To investigate the effects of clopidogrel on behavioral,content of inflammatory cytokines,and microglial in mice with comorbidity of pain and depression.Methods Twenty-four male pathogen-free C57BL/6J mice,aged 8 weeks,weighing 23-27 g,were selected.The mice were randomly divided into three groups:sham group(group C),spared nerve injury(SNI) group(group S),and SNI + clopidogrel group(group L),8 mice in each group.The surgical procedure in group C was consistent with the SNI model,but the nerve and maintained nerve integrity were not damaged.Group S was prepared with the SNI model,and group L was given clopidogrel 10 mg/kg since 21 days after the SNI model was prepared for 14 days continuously.Mechanical withdrawal threshold(MWT) was detected 1 day before surgery,7,14,21,28,and 35 days after surgery.The tail suspension test(TST) and forced swim test(FST) were performed 35 and 37 days after surgery,and the motionless time was calculated.Then the mice were killed,and the right hippocampal tissue was harvested to detect the concentration of IL-1β,IL-6,and TNF-α by enzyme-linked immunosorbent assay(ELISA),the expression of IL-1β,IL-6,and TNF-α mRNA by reverse transcription-polymerase chain reaction(RT-PCR),and the number of microglia by immunofluorescence staining.Results Compared with group C,MWT were significantly decreased 7,14,21,28,and 35 days after operation,the immobility time of TST and FST were significantly prolonged,the concentrations and mRNA expressions of IL-1β,IL-6,and TNF-α were significantly increased,and the number of microglia were significantly increased in groups S and L(P < 0.05).Compared with group S,MWT were significantly increased 28 and 35 days after operation,the immobility time of TST and FST were significantly shortened,the concentrations and mRNA expressions of IL-1β,IL-6,and TNF-α were significantly decreased,and the number of microglia was significantly reduced in group L(P < 0.05).Conclusion Clopidogrel can reduce the concentrations and mRNA expressions of IL-1β,IL-6,and TNF-α,inhibit the inflammatory reaction,reduce the number of activated microglia,and improve the and chronic neuropathic pain and depression in mice with comorbidity of pain and depression.