百合地黄汤抗抑郁作用机制初探

Preliminary Exploration of Antidepressant Mechanisms of Baihe Dihuang Decoction

目的探讨百合地黄汤对慢性不可预知应激(CUS)抑郁小鼠的抗抑郁作用及机制。方法将60只ICR小鼠根据糖水偏嗜度随机分为空白组10只和造模组50只,造模组再随机分为模型组、咯利普兰组、低剂量组、中剂量组、高剂量组各10只。造模组采用CUS法制备小鼠抑郁模型,造模周期35 d;空白组正常饲养。低、中、高剂量组每日按体质量20 mL/kg灌胃0.3、0.6、1.2 g/mL百合地黄汤药液,空白组及模型组每日按体质量20 mL/kg灌胃生理盐水,咯利普兰组每日按体质量10 mL/kg腹腔注射0.1 mg/mL咯利普兰药液,连续干预35 d。采用糖水偏好试验、旷场试验、强迫游泳试验、新奇抑制摄食试验、大理石埋藏试验、高架十字迷宫试验和Y迷宫空间试验检测6组小鼠行为学变化;ELISA法检测6组小鼠血清促肾上腺皮质激素(ACTH)、皮质酮(CORT)含量以及海马组织中cAMP含量。另将50只ICR小鼠根据体质量随机分为空白组、咯利普兰组、低剂量组、中剂量组、高剂量组各10只,5组给药方法和剂量同上,给药1 h后按体质量10 mL/kg腹腔注射1%戊巴比妥钠麻醉小鼠,观察5组翻正反射恢复时间结果与空白组比较,模型组糖水偏好试验的糖水偏嗜度明显降低,旷场试验的水平和垂直次数明显减少,强迫游泳试验的游泳不动时间增加,新奇抑制摄食试验的摄食潜伏期延长,大理石埋藏试验的埋藏弹珠数目增加,高架十字迷宫试验的开臂时间百分比、开臂次数百分比及入臂总次数下降,Y迷宫空间试验的交替反应率下降(P均<0.05),血清ACTH和CORT含量明显升高,海马组织中cAMP含量明显降低(P均<0.05);与模型组比较,咯利普兰组和各剂量组上述行为学指标均有改善,血清ACTH和CORT含量明显下降,海马组织中cAMP含量明显升高(P均<0.05);与空白组比较,咯利普兰组翻正反射恢复时间明显减少(P<0.05),各剂量组翻正反射恢复时间均无明显变化(P均>0.05)。结论百合地黄汤有显著的抗抑郁作用,且没有咯利普兰导致的呕吐不良反应,其作用机制与抑制ACTH和CORT含量和提升小鼠海马cAMP含量有关。

Objective:To investigate the antidepressant effect and mechanism of Baihe Dihuang decoction in mice with chronic unpredictable stress(CUS).Methods:A total of 60 male ICR mice were divided into control group(n=10)and model group(n=50)according to sucrose preference.The model group were randomly divided into CUS group,Rolipram group(0.1 mg/mL)and Baihe Dihuang decoction low,medium and high dose groups(0.3,0.6,1.2 g/mL).The mice were subjected to the CUS protocol for a period of 35 days to induce depressive-like behavior,while the control group was kept under normal conditions.Then,sucrose preference test,open-field test,forced swimming test,novelty-suppressed feeding test,marble burying test,elevated plus maze test and Y-maze test were performed to detect the antidepressant effects.Adrenocorticotropic hormone(ACTH)),corticosterone(CORT)in serum and cAMP expression in hippocampus were detected by enzyme-linked immunosorbent assay(ELISA)after the intervention lasted for 35 days.Then another 50 ICR mice were randomly divided into control group,Rolipram group(0.1 mg/mL)and Baihe Dihuang decoction low,medium and high dose groups(0.3,0.6,1.2 g/mL).The recovery time of righting reflex was observed 1 hour after intraperitoneal injection of 1% Pentobarbital sodium according to the body weight(10 mL/kg).Results:Compared with the control group,the sucrose preference significantly decreased,the number of crossings and rearings in open-field test significantly reduced,immobility time in forced swimming test significantly reduced,the feeding latency of novelty inhibition prolonged,the number of marbles buried increased,the time spent in open arms percentage,entries to open arms percentage and total arms entries in elevated plus maze test decreased,the spontaneous alternation in Y maze decreased(P<0.05),the ACTH and CORT level significantly increased,and cAMP expression levels in hippocampus significantly decreased(P<0.05)in the CUS group.As compared with the CUS group,the Rolipram group and the Baihe Dihuang decoction groups improved the aforementioned behavioral indicators,decreased the levels of ACTH and CORT significantly,while increased cAMP expression in hippocampus(P<0.05);compared with the control group,the recovery time of righting reflex of mice in the Rolipram group significantly decreaced(P<0.05),which didn't change significantly in the Baihe Dihuang decoction group(P>0.05).Conclusion:Baihe Dihuang decoction produced antidepressant effects in chronic unpredictable stress mice,and it had no adverse reaction of vomitting which can be caused by Rolipram.Its mechanism may be associated with inhibiting HPA axis activity and up-regulating cAMP expression in hippocampus.