松郁安神方通过cAMP/CREB信号通路改善失眠大鼠睡眠的研究

Songyu Anshen Formula Improved the Sleep of Insomnia Rats Through cAMP/CREB Signaling Pathway

目的 探讨松郁安神方(甘松、郁金、玫瑰花等)通过环磷酸腺苷(cAMP)/cAMP反应元件结合蛋白(CREB)信号通路改善失眠大鼠睡眠的作用机制。方法 将大鼠随机分为对照组、模型组及松郁安神方低、高剂量组。除对照组外,其余3组采用腹腔注射对氯苯丙氨酸(PCPA)复制失眠模型。模型复制成功后,松郁安神方低、高剂量组分别给予8.5、17 g·kg^(-1)松郁安神方灌胃,每天1次,连续7 d。观察大鼠一般状态,动物睡眠生物解析系统记录大鼠脑电(EEG)和肌电(EMG),采用ELISA法检测下丘脑cAMP含量,实时荧光定量PCR和Western Blot法分别检测下丘脑CREB mRNA和p-CREB蛋白表达。结果 与对照组比较,模型组大鼠白天时段非快速眼动(NREM)和快速眼动(REM)睡眠量均明显减少(P<0.01),觉醒量明显增加(P<0.01),下丘脑cAMP含量、CREB mRNA和p-CREB蛋白表达均明显降低(P<0.01)。松郁安神方治疗后,与模型组比较,松郁安神方低、高剂量组非快速眼动睡眠量均明显增加(P<0.05,P<0.01),觉醒量均明显减少(P<0.05,P<0.01),下丘脑c AMP含量、CREB mRNA和p-CREB蛋白表达均明显升高(P<0.05,P<0.01)。结论 松郁安神方可能通过下丘脑c AMP/CREB信号通路改善失眠大鼠睡眠。

Objective To research the effects of Songyu Anshen Formula(SYF;Nardostachyos Radix et Rhizoma,Curcumae Radix,Rosae Rugosae Flos,etc)on cAMP/CREB signaling pathway in hypothalamus of insomnia rats.Methods The rats were randomly divided into control group,model group,low-dose SYF group and high-dose SYF group. Except the control group, the rat model of insomnia of the other groups was established by intraperitoneal injection with para-chlorophenylalanine(PCPA). After the model was copied successfully, the rats were intragastrically administrated with SYF(8.5,17 g·kg^(-1),respectively)once daily for 7 days. The general state of rats was observed during administration. The changes of sleep in rats were observed by recording electroencephalogram(EEG) and electromyogram(EMG). The cAMP content in hypothalamus was detected by ELISA. CREB m RNA and p-CREB protein expression were determined by RT-PCR and Western Blot,respectively.Results Compared with the control group, the amount of non-rapid-eye-movement(NREM) and rapid-eyemovement(REM)daytime sleep in model group was significantly reduced(P<0.01),and the amount of wakefulness was significantly increased(P<0.01). The content of c AMP,CREB mRNA and p-CREB protein in hypothalamus were significantly reduced(P<0.01). Compared with model group,the amount of NREM and REM sleep in lowand high-dose SYF groups was obviously increased(P<0.05, P<0.01) after administration of SYF, and the amount of wakefulness was decreased(P<0.05, P<0.01). The content of cAMP, CREB mRNA and p-CREB protein in hypothalamus were significantly increased(P<0.05,P<0.01). Conclusion SYF may improve sleep of insomniac rats through cAMP/CREB signaling pathway in the hypothalamus.