本研究的目的是研究雌激素受体β(ERβ)在壬基酚(NP)诱导的大鼠抑郁样行为中的作用及其对TPH2/5-HT通路调节的影响。在体外实验中,大鼠嗜碱性白血病细胞(RBL-2H3)分为4组:空白组,NP组(20μM),ERβ激动剂组(0.01μM),NP+ERβ激动剂组(20μM+0.01μM)。对于体内实验,72只成年雄性SD大鼠随机分为6组:对照组,NP(40mg/kg)组,ERβ激动剂(2mg/kg,二芳基丙腈(DPN)基团,ERβ抑制剂(0.1mg/kg,4-(2-苯基-5,7-双(三氟甲基)吡唑并[1,5-a]嘧啶-3-基)苯酚(PHTPP)基团,NP+ERβ激动剂(40mg/kgNP+2mg/kgDPN)组,和NP+ERβ抑制剂(40mg/kgNP+0.1mg/kgPHTPP)组,每组12只大鼠。药物组的每只大鼠通过管饲法给予NP和/或单次腹膜内注射DPN2mg/kg或PHTPP0.1mg/kg。在体内和体外,NP组显示ERβ表达水平降低,色氨酸羟化酶(TPH1),和色氨酸羟化酶-2(TPH2)基因和蛋白质,降低DA水平,NE,和5-羟色氨酸(5-HT)神经递质。RBL-2H3细胞显示细胞收缩的迹象,圆形细胞,增加悬浮和更松散排列的细胞。ERβ激动剂刺激的有效性在RBL-2H3细胞中表现出超过60%的增加。ERβ激动剂的应用导致上述改变的缓解。ERβ激动剂激活TPH2/5-HT信号通路。与对照组相比,NP组脑组织中NP含量显著增高。大鼠进食的潜伏期较长,消耗的食物量较低,在大鼠行为实验中,大鼠的不动时间延长。ERβ的表达水平,NP组TPH1、TPH2、5-HT和5-HITT蛋白均降低,提示NP诱导的抑郁样行为以及血清激素和单胺类神经递质分泌的紊乱。在NP组中,中线中缝核显示出细长的核,呈深紫蓝色,核萎缩,位移和苍白的细胞质。ERβ可能改善NP诱导的抑郁样行为,和分泌障碍的血清激素和单胺类神经递质通过激活TPH2/5-HT信号通路。
The aim of this study is to investigate the role of estrogen receptor β (ERβ) in
nonylphenol (NP) - induced depression - like behavior in rats and its impact on the regulation of the TPH2/5-HT pathway. In the in vitro experiment, rat basophilic leukaemia cells (RBL-2H3) cells were divided into the four groups: blank group, NP group (20 μM), ERβ agonist group (0.01 μM), and NP+ERβ agonist group (20 μM+0.01 μM). For the in vivo experiment, 72 adult male Sprague-Dawley rats were randomly divided into following six groups: the Control, NP (40 mg/kg) group, ERβ agonist (2 mg/kg, Diarylpropionitrile (DPN)) group, ERβ inhibitor (0.1 mg/kg, 4-(2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl) phenol (PHTPP)) group, NP+ERβ agonist (40 mg/kg NP + 2 mg/kg DPN) group, and NP+ERβ inhibitor (40 mg/kg NP + 0.1 mg/kg PHTPP) group, with 12 rats in each group. Each rat in drug group were given NP by gavage and/or received a single intraperitoneal injection of DPN 2 mg/kg or PHTPP 0.1 mg/kg. Both in vivo and in vitro, NP group showed a decrease in the expression levels of ERβ, tryptophan hydroxylase (TPH1), and tryptophan hydroxylase-2 (TPH2) genes and proteins, and reduced levels of DA, NE, and 5-hydroxytryptophan (5-HT) neurotransmitters. RBL-2H3 cells showed signs of cell shrinkage, with rounded cells, increased suspension and more loosely arranged cells. The effectiveness of the ERβ agonist stimulation exhibited an increase exceeding 60% in RBL-2H3 cells. The application of ERβ agonist resulted in an alleviation the aforementioned alterations. ERβ agonist activated the TPH2/5-HT signaling pathways. Compared to the control group, the NP content in the brain tissue of the NP group was significantly increased. The latency to eat for the rats was longer and the amount of food consumed was lower, and the rats had prolonged immobility time in the behavioral experiment of rats. The expression levels of ERβ, TPH1, TPH2, 5-HT and 5-HITT proteins were decreased in the NP group, suggesting NP-induced depression-like behaviours as well as disturbances in the secretion of serum hormones and monoamine neurotransmitters. In the NP group, the midline raphe nucleus showed an elongated nucleus with a dark purplish-blue colour, nuclear atrophy, displacement and pale cytoplasm. ERβ might ameliorate NP-induced depression-like behaviors, and secretion disorders of serum hormones and monoamine neurotransmitters via activating TPH2/5-HT signaling pathways.