睡眠剥夺对小鼠大肠组织炎症反应和氧化应激的影响

Effects of sleep deprivation on intestinal inflammatory response and oxidative stress in mice

目的探讨睡眠剥夺对小鼠大肠组织炎症反应和氧化应激的影响。方法选择SPF级雄性C57小鼠32只,8~12周龄,体重18~22 g。采用随机数字表法分为四组:空白对照组(C组)、切口组(I组)、睡眠剥夺组(A组)和睡眠剥夺+切口组(AI组),每组8只。C组正常饲养,I组建立切口模型,A组使用睡眠剥夺箱睡眠剥夺48 h,AI组使用睡眠剥夺箱睡眠剥夺48 h后建立切口模型。取小鼠大肠组织,采用Western blot法检测白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(NOX2)和超氧化物歧化酶2(SOD2)蛋白含量,超氧化物阴离子荧光探针法检测活性氧(ROS)荧光强度,免疫荧光染色法检测NOX2荧光强度。结果与C组比较,I组、A组和AI组大肠组织IL-1β、TNF-α和NOX2蛋白含量明显升高(P<0.05),SOD2蛋白含量明显降低(P<0.05),ROS和NOX2荧光强度明显增强(P<0.05)。与I组比较,A组和AI组大肠组织IL-1β、TNF-α和NOX2蛋白含量明显升高(P<0.05),AI组大肠组织SOD2蛋白含量明显降低(P<0.05),ROS和NOX2荧光强度明显增强(P<0.05)。与A组比较,AI组大肠组织IL-1β、TNF-α和NOX2蛋白含量明显升高(P<0.05),SOD2蛋白含量明显降低(P<0.05),ROS和NOX2荧光强度明显增强(P<0.05)。结论睡眠剥夺会引起小鼠大肠组织炎症反应和氧化应激,睡眠剥夺下行切口手术会进一步加重炎症反应与氧化应激。

Objective To investigate the effects of sleep deprivation on intestinal oxidative stress and inflammatory response in mice.Methods Thirty-two SPF male C57 mice,aged 8-12 weeks,weighing 18-22 g,were divided into four groups using random number table method:blank control group(group C),incision group(group I),sleep deprivation group(group A),and sleep deprivation+incision group(group AI),eight mice in each group.Group C were kept normally,group I were established into incision model,group A were established into sleep deprivation model by depriving for 48 hours in a sleep deprivation chamber,and group AI were established into incision and sleep deprived models.The expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),nicotinamide adenine dinucleotide phosphate oxidase 2(NOX2)and superoxide dismutase 2(SOD2)protein were detected by Western blot.The fluorescence intensity of reactive oxygen species(ROS)was detected bydihydroethidium and NOX2 fluorescence intensity was detected by immunofluorescence.Results Compared with group C,the expression of IL-1β,TNF-αand NOX2 protein were significantly increased(P<0.05),the expression of SOD2 protein was significantly decreased(P<0.05),and ROS and NOX2 fluorescence intensity were significantly enhanced(P<0.05)in colonic tissues in groups I,A and AI.Compared with group I,the expression of IL-1β,TNF-αand NOX2 protein of colonic tissues in groups A and AI were significantly increased(P<0.05),the expression of SOD2 protein in group AI was significantly decreased(P<0.05),and the fluorescence intensity of ROS and NOX2 in group AI were significantly enhanced(P<0.05).Compared with group A,the expression of IL-1β,TNF-αand NOX2 protein were significantly increased(P<0.05),the expression of SOD2 protein was significantly decreased(P<0.05),and ROS and NOX2 fluorescence intensity were significantly enhanced(P<0.05)in colonic tissues in group AI.Conclusion Sleep deprivation can cause intestinal inflammatory response and oxidative stress in mice,and the inflammatory response and oxidative stress will be further aggravated by sleep deprivation and surgery.