Bai Yongtao, Dai Guoliang, Song Lihua, Gu Xiaolei, Ba Ning, Ju Wenzheng, Zhang Wenzhou
Department of Pharmacy, The Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou, China.
Clinical Research Center, The Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou, China.
Front Pharmacol. 2022 Jul 6;13:918776. doi: 10.3389/fphar.2022.918776. eCollection 2022.
Zhi-Zi Hou-Po Decoction (ZHD) has been widely used in the treatment of depression for centuries. This study aimed to investigate the antidepressant effects of the water extract of ZHD (ZHD-WE) and ethanol extract of ZHD (ZHD-EE) using behavioral despair tests in mice, and to further explore the neuroprotective effects in a PC12 cell injury model induced by corticosterone (CORT). Mice were divided into a control group (normal saline), ZHD-WE groups (4, 8, and 16 g kg), ZHD-EE groups (4, 8, and 16 g kg) and the fluoxetine group (20 mg kg). The forced swimming test (FST) and tail suspension test (TST) were used to screen the antidepressant effects of ZHD-WE and ZHD-EE after oral administration for seven consecutive days. The level of brain-derived neurotrophic factor (BDNF) in the hippocampus was determined by ELISA. The MTT, lactate dehydrogenase (LDH) and flow cytometry analysis were performed to elucidate the neuroprotective effect of ZHD-EE on a PC12 cell injury model. Additionally, the mRNA and proteins expression of apoptotic molecules Bax, Bcl-2 and BDNF were detected by RT-PCR and western blot assay. It showed that ZHD-EE at concentrations of 8 and 16 g kg significantly decreased the immobility time in the TST and FST, and increased the BDNF levels in the hippocampus. While ZHD-WE at concentrations of 4, 8, and 16 g kg had no significant effect on the immobility time in the TST, and only the 16 g kg of extract group significantly decreased the immobility time in the FST. the obtained results showed that PC12 cells pre-incubated with ZHD-EE at concentrations of 100 and 400 μg ml improved cell viability, decreased LDH release, and reduced apoptosis rate of PC12 cells. Moreover, ZHD-EE significantly increased the mRNA and proteins expression of Bcl-2 and BDNF, while decreased the mRNA and protein expression of Bax. ZHD-EE significantly improved despair-like behavior in mice, and its mechanism may be related to BDNF upregulation in the hippocampus. This study also showed that ZHD-EE had a protective effect on CORT-induced injury in PC12 cells by upregulating the expression of BDNF and restoring Bcl-2/Bax balance.
栀子厚朴汤（ZHD）几个世纪以来一直广泛用于治疗抑郁症。本研究旨在通过小鼠行为绝望试验研究ZHD水提取物（ZHD-WE）和ZHD乙醇提取物（ZHD-EE）的抗抑郁作用，并在皮质酮（CORT）诱导的PC12细胞损伤模型中进一步探讨其神经保护作用。将小鼠分为对照组（生理盐水）、ZHD-WE组（4、8和16 g/kg）、ZHD-EE组（4、8和16 g/kg）和氟西汀组（20 mg/kg）。连续7天口服给药后，采用强迫游泳试验（FST）和悬尾试验（TST）筛选ZHD-WE和ZHD-EE的抗抑郁作用。通过ELISA法测定海马中脑源性神经营养因子（BDNF）水平。进行MTT、乳酸脱氢酶（LDH）和流式细胞术分析以阐明ZHD-EE对PC12细胞损伤模型的神经保护作用。此外，通过RT-PCR和蛋白质印迹法检测凋亡分子Bax、Bcl-2和BDNF的mRNA和蛋白质表达。结果显示，8和16 g/kg浓度的ZHD-EE显著缩短了TST和FST中的不动时间，并提高了海马中的BDNF水平。而4、8和16 g/kg浓度的ZHD-WE对TST中的不动时间无显著影响，仅16 g/kg提取物组显著缩短了FST中的不动时间。结果表明，用100和400 μg/ml浓度的ZHD-EE预孵育的PC12细胞提高了细胞活力，降低了LDH释放，并降低了PC12细胞的凋亡率。此外，ZHD-EE显著增加了Bcl-2和BDNF的mRNA和蛋白质表达，同时降低了Bax的mRNA和蛋白质表达。ZHD-EE显著改善了小鼠的绝望样行为，其机制可能与海马中BDNF上调有关。本研究还表明，ZHD-EE通过上调BDNF表达和恢复Bcl-2/Bax平衡对CORT诱导的PC12细胞损伤具有保护作用。
