电针干预对慢性睡眠剥夺大鼠伏隔核ANXA1、IL-1β表达的影响

Effects of EA Intervention on Expressions of ANXA1 and IL-1βin Nucleus Accumbens of Rats with Chronic Sleep Deprivation

目的:观察电针对慢性睡眠剥夺(CSD)大鼠伏隔核(NAc)膜联蛋白A1(ANXA1)、白细胞介素1β(IL-1β)炎症因子的调节作用,分析电针改善睡眠及焦虑样行为的可能机制。方法:48只雄性SD大鼠随机分为空白组、模型组、电针组与假针刺组4组,每组12只。采用改良多平台水环境法(MMPM)建立CSD大鼠模型。电针组选用单日“百会”“神门”和双日双侧“内关”“太冲”电针干预21 d。实验过程中观察大鼠一般情况及体质量变化;旷场实验观察大鼠自主活动能力,评价焦虑程度;实时荧光定量PCR检测大鼠NAc脑区ANXA1、IL-1βmRNA表达水平,蛋白免疫印记(WB)检测NAc中ANXA1、IL-1β蛋白表达水平。结果:21 d造模后,与空白组比较,其余3组大鼠毛发干枯无光泽,体质量增长缓慢(P<0.01);大鼠在旷场中运动总距离及平均速度增加(P<0.05或P<0.01)、中央区停留时间减少(P<0.01)。21 d干预后,与同时段空白组比较,模型组大鼠体质量依旧增长缓慢(P<0.01),旷场实验中大鼠运动总距离及平均速度增加(P<0.01)、中央区停留时间减少(P<0.05),IL-1βmRNA和蛋白表达水平均明显降低(P<0.01)、ANXA1升高(P<0.01);与模型组比较,电针组大鼠体质量开始逐渐上升(P<0.01),精神状态良好,大鼠运动总距离、平均速度降低(P<0.05或P<0.01),中央区停留时间增加(P<0.05),IL-1β在mRNA和蛋白表达水平上均显著升高、ANXA1明显降低(P<0.01);假针刺组与模型组比较,差异均无统计学意义(P>0.05)。结论:电针可有效改善CSD大鼠睡眠及焦虑状态,其机制可能与升高CSD大鼠NAc脑区IL-1β、降低ANXA1炎症因子的表达有关。

Objective:To observe the regulatory effect of electro-acupuncture(EA)on the expressions of pro-inflammatory cytokines annexin A1(ANXA1)and interleukin 1β(IL-1β)in the nucleus accumbens(NAc)of rats with chronic sleep deprivation(CSD),and to analyze the potential mechanisms of EA in improving sleep and anxiety-like behaviors.Methods:48 male SD rats were randomly divided into the control group,the model group,the EA group,and the sham acupuncture group,with 12 rats in each group.The modified multi-platform method(MMPM)was used to establish the CSD rat model.The EA group was treated with electro-needling Baihui(DU20)and Shenmen(HT7)on the odd-numbered days,and electro-needling Neiguan(PC6)and Taichong(LR3)on the even-numbered days,and the EA intervention duration was 21 days.The general condition and body weight changes were observed.The open field test was conducted to evaluate the spontaneous activity and anxiety levels.The real-time quantitative PCR was used to detect the mRNA expressions of ANXA1 and IL-1βin the NAc brain region.The Western Blot was used to detect the protein expressions of ANXA1 and IL-1βin the NAc brain region.Results:After 21 days of modeling,compared with those in the control group,the rats in the rest three groups had dry and dull hair with slow increased weight(P<0.01);the total distance and the average speed of movement in the open field increased(P<0.05,P<0.01),and the time spent in the central area decreased(P<0.01).After 21 days of intervention,compared with those in the control group,the weight gain was slow(P<0.01),the total distance and the average speed of movement in the open field were increased(P<0.01),the time spent in the central area was decreased(P<0.05),the mRNA and protein expressions of IL-1βwere significantly decreased(P<0.01),whereas the mRNA and protein expresssions of ANXA1 were increased in the model group at the same period(P<0.01).Compared with those in the model group,the body weight was gradually increased(P<0.01),the mental state was good,the total distance and the average speed of movement were decreased(P<0.05,P<0.01),the time spent in the central area was increased(P<0.05),the mRNA and protein expressions of IL-1βwere significantly increased,whereas the mRNA and protein expressions of ANXA1 were significantly decreased in the EA group(P<0.01).There were no statistical differences between the sham acupuncture group and the model group(P>0.05).Conclusion:EA can effectively improve sleep and regulate anxiety status of CSD rats,which mechanism may be related to up-regulating IL-1βand down-regulating ANXA1 in the NAc brain region of CSD rats.