交泰丸对睡眠剥夺小鼠视交叉上核生物钟基因Clock及Bmal 1表达的影响

Effects of Jiaotai Pills on the Expression of Clock and Bmal 1 in the Suprachiasmatic Nucleus of Sleep Deprived Mice

目的观察交泰丸对睡眠剥夺小鼠视交叉上核(SCN)中生物钟基因Clock和Bmal 1表达的影响,探讨交泰丸改善睡眠的可能机制。方法将小鼠随机分为空白组、模型组、百乐眠组和交泰丸组。除空白组外,其他各组腹腔注射氯苯丙氨酸(PCPA)以制备睡眠剥夺模型。用自主活动测试仪记录小鼠自主活动时间,ELISA检测血清中5-HT、Glu、DA、GABA的表达水平,Real time-PCR和免疫组化检测SCN中Clock和Bmal 1的表达。结果与空白组比较,模型组小鼠活动时间显著增加(<0.01);与模型组比较,百乐眠组和交泰丸组小鼠活动时间有不同程度减少(<0.05)。与空白组比较,模型组小鼠血清中GABA、5-HT含量降低(<0.01),Glu含量升高(<0.01);与模型组比较,百乐眠组和交泰丸组小鼠血清中GABA、5-HT含量升高(<0.01,<0.05),Glu含量降低(<0.05);各组小鼠血清中DA含量变化不明显,差异没有统计学意义(>0.05)。与空白组比较,模型组小鼠SCN中Clock、Bmal1的基因和蛋白表达水平升高(<0.01);与模型组比较,百乐眠组和交泰丸组小鼠SCN中Clock、Bmal1的基因和蛋白表达下降(<0.01,<0.05)。结论交泰丸具有改善睡眠的作用,其机制可能与交泰丸调节小鼠SCN中Clock和Bmal 1的表达有关。

Objective To observe the effect of Jiaotai pills on the expression of Clock and Bmal1 in suprachiasmatic nucleus(SCN)of sleep deprived mice,and to explore its possible mechanism in improving sleeping.Methods Mice were randomly divided into blank group,model group,Bailemian capsule group and Jiaotai pills group.All groups were injected with chlorophenylalanine(PCPA)intraperitoneally to establish sleep deprivation model except blank group.Autonomous activity time of mice was recorded by Autonomous Activity Tester.ELISA was used to detect 5-HT,Glu,DA and GABA levels in serum.Real time-PCR and Immunohistochemistry were used to detect the expression of Clock and Bmal 1 in SCN.Results Compared with blank group,the activity time of mice in model group increased significantly(<0.01);compared with model group,the activity time of mice in Bailemian capsule group and Jiaotai pills group decreased in varying degrees(<0.05).Compared with blank group,the levels of GABA and 5-HT in the serum of model group decreased(<0.01),while the levels of Glu increased(<0.01);compared with model group,the levels of GABA and 5-HT in the serum of Bailemian capsule group and Jiaotai pillsgroup increased(<0.01,<0.05),and the levels of Glu decreased(<0.05);there was no change in the levels of DA in the serum of mice in each group(>0.05).Compared with blank group,the gene and protein expression levels of Clock and Bmal1 in SCN of mice in model group increased(<0.01);compared with model group,the gene and protein expression levels of Clock and Bmal1 in SCN of mice in Bailemian capsule and Jiaotai pills groups decreased(<0.01,<0.05).Conclusion Jiaotai pills can improve sleeping,its mechanism may be related to the regulation of Clock and Bmal 1 expression in SCN of mice.