佛手宁神方改善睡眠作用的机制

Foshouningshen decoction improves sleeping via the serotonergic system in a rat model of insomnia

目的评价佛手宁神方镇静催眠作用并研究其对失眠大鼠海马5-羟色胺(5-HT)、5-羟色胺1A受体(5-HT1AR)表达的影响。方法雄性昆明小鼠随机分为空白组,艾司唑仑组[0.4 mg/(kg·d)],佛手宁神方低、中、高剂量组[12,24,48 g/(kg·d)];连续给药1周,末次给药后进行阈下剂量和阈剂量戊巴比妥钠致小鼠睡眠实验。雄性SD大鼠随机分为空白组,模型组,艾司唑仑组[0.2 mg/(kg·d)],佛手宁神方低、中、高剂量组[6,12,24 g/(kg·d)],每组8只;采用腹腔注射对氯苯丙氨酸(PCPA)350 mg/(kg·d)制作失眠大鼠模型;造模完成后第1天开始灌胃给药,连续1周。旷场实验测试各组大鼠运动总路程和站立次数;酶联免疫吸附法测定海马5-HT的含量;免疫组化法和实时荧光定量PCR分别检测海马5-HT1AR蛋白和m RNA的表达。结果与空白组相比,佛手宁神方高剂量组阈下剂量戊巴比妥钠致小鼠入睡只数增加(P<0.05),阈剂量戊巴比妥钠致小鼠睡眠潜伏期缩短(P<0.01),睡眠持续时间延长(P<0.01)。与空白组比较,模型组大鼠在旷场实验中运动总路程增加(P<0.05),海马5-HT含量减少(P<0.01),海马5-HT1AR蛋白和m RNA表达水平下降(P<0.01)。与模型组相比,艾司唑仑组和佛手宁神方高剂量组运功总路程减少(P<0.05),海马5-HT含量增加(P<0.05),海马5-HT1AR蛋白和m RNA表达水平上升(P<0.01)。结论佛手宁神方可能通过提高海马5-HT的含量,上调海马5-HT1AR蛋白和m RNA的表达,从而起到治疗失眠的作用。

Objective To evaluate the sedative and hypnotic effects of Foshouningshen decoction （FSNSD） and study its effects on expressions of 5-hydroxy tryptamine （5-HT） and 5-HT1A receptor （5-HT1AR） in the hippocampus in a rat model of insomnia. Methods Male KM mice were divided into control group, estazolam （0.4 mg/kg daily） group, and low-, moderate-, and high-dose FSNSD groups （daily dose of 12, 24, and 48 g/kg, respectively）. After corresponding treatments for 1 week, the mice underwent sleep-inducing test with subthreshold and threshold doses of sodium pentobarbital. Forty-eight male SD rats were randomized into control group, insomnia model group, estazolam group （0.2 mg/kg daily）, and low-, moderate-, and high-dose FSNSD groups （with daily dose of 6, 12, and 24 g/kg, respectively）. Rat models of insomnia were established by intraperitoneal injection of 4-cholro-dl-phenylalanine （PCPA） at the daily dose of 350 mg/kg for 3 days, after which the rats received corresponding treatments via gavage for 1 week. The performance of the rats in open field test was recorded and the hippocampal expression of 5-HT was detected using ELISA; the expressions of 5-HT1AR protein and mRNA in the hippocampus were detected using immunohistochemistry and real-time PCR, respectively. Results In the sleep-inducing test with a subthreshold dose of sodium pentobarbital, the mice treated with high-dose FSNSD showed a significantly higher rate of sleep onset than the control mice （P〈0.05）; in the test with a threshold dose of sodium pentobarbital, treatment with moderate- and high-dose FSNSD resulted in significantly prolonged sleeping time （P〈0.01） and shortened sleep latency （P〈0.05） in the mice. The rats in insomnia model group showed increased total distance in open field test （P〈0.05） with significantly decreased content of 5-HT （P〈0.01） and expressions of 5-HT1AR protein and mRNA in the hippocampus （P〈0.01）. Treatment of the rats with estazolam or high-dose FSNSD obviously decreased the total distance in open field test （P〈0.05） and increased the content of 5-HT （P〈0.05） and expressions of 5-HT1AR （P〈0.01） in the hippocampus of rats with insomnia. Conclusion FSNSD can produce therapeutic effects on insomnia possibly by increasing 5-HT content and expressions of 5-HT1AR in the hippocampus.