Cao Kerun, Shen Chongkun, Yuan Yumei, Bai Shasha, Yang Lei, Guo Lili, Zhang Rong, Shi Yafei
School of Fundamental Medical Science, Guangzhou University of Chinese Medicine, Guangzhou, China.
Shenzhen Baoan Hospital of Chinese Medicine, Shenzhen, China.
Front Psychiatry. 2019 Mar 28;10:160. doi: 10.3389/fpsyt.2019.00160. eCollection 2019.
Early adverse life stress is an important dangerous factor in the development of psychiatric disorders, particularly depression. Available clinical antidepressant agents, such as fluoxetine, [a selective serotonin reuptake inhibitor (SSRI)], are unsatisfactory because of their side effects. SiNiSan (SNS) is a classic Chinese medicine prescription regarded to disperse stagnated liver qi to relieve qi stagnation. Therefore, this study was designed to detect the effects and molecular mechanism of SNS treatment in rats subjected to maternal separation (MS). Male neonatal Wistar rats were divided into six groups including control + ddHO, MS + ddHO, MS + fluoxetine (5 g/kg), MS + SNS -low dose (2.5 g/kg), MS + SNS -medium dose (5 g/kg), MS + SNS -high dose (10 g/kg). The volume of drugs and ddH2O in each group are according to the weight of rats every day (10 mL/kg). Each group comprised 16 pups with 8 young and 8 adult pups. Except for the control group, all MS groups were separated from their mothers for 4 h/day from 9:00 to 13:00 during postnatal days (PNDs) 1 to 21. After MS, the six groups were intragastrically administered with ddH2O, fluoxetine, and different doses of SNS until PND 28 (for young pups) and PND 56 (for adult pups). The pups were weighed every day, and depression-like behavior was assessed by sucrose preference test, open field test, and forced swimming test. Serotonin 1A (5-HT1A) receptor, phosphorylated protein kinase A (p-PKA) substrate, cAMP response element-binding protein (CREB), p-CREB and brain-derived neurotrophic factor (BDNF) in the hippocampus were examined by Western blot, and 5-HT1A receptor expression was measured by IHC. Young and adult MS rats exhibited depression-like behavior. However, the depression-like behavior was ameliorated by SNS in both age groups. The levels of 5-HT1A receptor, p-CREB, and BDNF in the hippocampus were reduced in young and adult MS rats. SNS treatment significantly up-regulated the expression of 5-HT1A receptor, p-CREB, and BDNF in the hippocampus of adult MS rats. However, few significant effects on the protein expression were observed in the young MS rats. MS in infancy could develop depression-like behavior in young and adult. SNS treatment may perform antidepressant effects on young and adult MS rats through the BDNF/PKA/CREB pathway.
早期不良生活应激是精神疾病尤其是抑郁症发生发展的重要危险因素。现有的临床抗抑郁药，如氟西汀（一种选择性5-羟色胺再摄取抑制剂，SSRI），由于其副作用而不尽人意。四逆散（SNS）是一种经典的中药方剂，被认为具有疏肝理气、解郁的作用。因此，本研究旨在检测SNS对经历母婴分离（MS）大鼠的治疗效果及分子机制。雄性新生Wistar大鼠分为六组，包括对照组+双蒸水、MS+双蒸水、MS+氟西汀（5 g/kg）、MS+SNS低剂量组（2.5 g/kg）、MS+SNS中剂量组（5 g/kg）、MS+SNS高剂量组（10 g/kg）。每组药物和双蒸水的体积根据大鼠体重每天给予（10 mL/kg）。每组包含16只幼崽，其中8只为幼年，8只为成年。除对照组外，所有MS组在出生后第1至21天，每天从9:00至13:00与母亲分离4小时。MS后，六组分别灌胃给予双蒸水、氟西汀和不同剂量的SNS，直至出生后第28天（幼年大鼠）和第56天（成年大鼠）。每天测量幼崽体重，并通过蔗糖偏好试验、旷场试验和强迫游泳试验评估抑郁样行为。通过蛋白质免疫印迹法检测海马中5-羟色胺1A（5-HT1A）受体、磷酸化蛋白激酶A（p-PKA）底物、cAMP反应元件结合蛋白（CREB）、磷酸化CREB（p-CREB）和脑源性神经营养因子（BDNF），并通过免疫组化法检测5-HT I A受体表达。幼年和成年MS大鼠均表现出抑郁样行为。然而，两个年龄组的抑郁样行为均被SNS改善。幼年和成年MS大鼠海马中5-HT1A受体、p-CREB和BDNF水平降低。SNS治疗显著上调成年MS大鼠海马中5-HT1A受体、p-CREB和BDNF的表达。然而，在幼年MS大鼠中未观察到对蛋白质表达的显著影响。幼年时期的MS可使幼年和成年大鼠出现抑郁样行为。SNS治疗可能通过BDNF/PKA/CREB途径对幼年和成年MS大鼠发挥抗抑郁作用。
