miR-373通过P2X7R影响抑郁症小鼠行为的作用机制

Mechanism of miR-373 on Behaviors of Depression-mice via P2X7R

目的探讨miR-373对抑郁症小鼠模型抑郁样行为,小胶质细胞激活和焦亡的影响。方法采用慢性不可预知应激构建抑郁症小鼠模型。蔗糖偏好,强迫游泳,尾部悬挂和社会实验评估小鼠的抑郁样行为。免疫荧光双染小胶质细胞标志物Iba-1和小胶质细胞活化标志物OX-42以评估小鼠海马层中小胶质细胞增殖和活化状态。TUNEL试剂盒检测小胶质细胞的凋亡。荧光定量PCR检测miR-373的表达水平。蛋白质印记检测P2X7R和细胞焦亡相关蛋白的表达水平。双荧光素酶报告基因实验验证miR-373和P2X7R的靶向关系。结果慢性不可预知应激处理的小鼠,蔗糖偏好度和社交时间显著下调,并且强迫游泳和尾部悬挂不动时间显著增加(P<0.05)。miR-373在抑郁症模型小鼠中异常高表达,且能够缓解小鼠的抑郁样行为(P<0.05)。miR-373靶向负调控小鼠海马层小胶质细胞中P2X7R的表达水平(P<0.01)。miR-373抑制小鼠海马层中小胶质细胞增殖和激活(P<0.01)。miR-373抑制小胶质细胞中Caspase-1,C-caspase-1,NLRP3,IL-1β和IL-18表达,并抑制小胶质细胞凋亡(P<0.05)。结论miR-373通过靶向抑制P2X7R的表达,从而缓解抑郁症小鼠模型的抑郁样行为,并抑制其海马层中小胶质细胞活化和焦亡。

Objective To explore the effect of miR-373 on depression-like behavior,microglia activation and pyrolysis in depression mice.Methods We Used chronic unpredictable stress to establish depression mice model.Sucrose preference,forced swimming,tail suspension,and Social interaction test were evaluate depressionlike behaviors.Immunofluorescence double-stained microglia marker Iba-1 and microglia activation marker OX-42 were used to evaluate the proliferation and activation of microglia in the hippocampus of mice.TUNEL kit was used to detect microglia apoptosis.RT-qPCR was used to detect miR-373 expression.Western blotting was used to detect the expression of P2 X7 R and pyrolysis-related proteins.The targeting relationship between miR-373 and P2 X7 R was verified by dual-luciferase reporter gene assay.Results In mice treated with chronic unpredictable stress,sucrose preference and social time were significantly reduced,and immobility time of forced swimming and tail suspension was significantly increased(P<0.05).miR-373 washighly expressed,and alleviated depression-like behavior in depression model mice(P<0.05).mi R-373 targeted and negatively regulated P2 X7 R expression in microglia ofmice hippocampus(P<0.01).mi R-373 inhibited the proliferation and activation of microglia(P<0.01).mi R-373 inhibited caspase-1,c-caspase-1,NLRP3,IL-1βand IL-18 expression,and the apoptosis of microglia(P<0.05).Conclusion Imi R-373 inhibits P2 X7 R expression to alleviate depression-like behavior,and inhibit the activation and pyroptosis of microglia in the hippocampus of depression mice.