Yu He, He Zhong-Mei, Shi Kun, Zhao Yan, Zong Ying, Li Jian-Ming, Chen Wei-Jia, DU Rui
College of Chinese Medicinal Materials,Jilin Agricultural University Changchun 130118,China.
College of Chinese Medicinal Materials,Jilin Agricultural University Changchun 130118,China Key Laboratory of Animal Production,Product Quality and Security,Ministry of Education of China Changchun 130118,China Jilin Provincial Engineering Research Center for Efficient Breeding and Product Development of Sika Deer of China Changchun 130118,China.
Zhongguo Zhong Yao Za Zhi. 2021 Jul;46(14):3678-3686. doi: 10.19540/j.cnki.cjcmm.20210406.403.
This study aimed to investigate the antidepressant effects of total alkaloids of Fibraurea recisa in HT22 cells damaged by corticosterone (CORT) in vitro and in a mouse model of chronic unpredictable mild stress (CUMS) as well as the underlying mechanisms.In cellular experiments,the viability of CORT-damaged HT22 cells was detected using cell counting kit-8 (CCK-8),and the cell apoptosis was detected by Hoechst 33258 staining.In animal experiments,C57BL/6N mice were randomly divided into the control group,model group,low (100 mg·kg(-1)),medium (200 mg·kg(-1)) and high (400 mg·kg~(-1))-dose of total alkaloids of F.recisa groups,and positive control group.After 21 days of CUMS exposure,their depressive behaviors were observed in behavioral and Morris water maze tests.The serum levels of 5-hydroxytryptamine (5-HT),dopamine (DA),and norepinephrine (NE) were assessed by ELISA.The expression levels of apoptosis-related proteins Bcl-2,Bax and cleaved caspase-3 in HT22 cells and mouse hippocampus were detected by Western blot.The results suggested that total alkaloids of F.recisa alleviated the damage of HT22 cells induced by CORT in a dose-dependent manner.The Hoechst 33258 staining uncovered that total alkaloids of F.recisa better reduced the blue spots and inhibited cell apoptosis.The results of animal experiments showed that total alkaloids of F.recisa significantly improved the depression-like behaviors of mice and increased the serum levels of 5-HT,DA and NE as compared with those in the model group.The Western blot assays revealed a significant up-regulation of Bcl-2 protein expression,but an obvious reduction in Bax and cleaved caspase-3protein expression in the total alkaloids of F.recisa group.In conclusion,total alkaloids of F.recisa inhibited depression possibly by regulating the apoptosis-related protein expression or elevating the monoamine neurotransmitter levels in the brain.
本研究旨在探讨黄藤总生物碱对体外皮质酮（CORT）损伤的HT22细胞以及慢性不可预测轻度应激（CUMS）小鼠模型的抗抑郁作用及其潜在机制。在细胞实验中，使用细胞计数试剂盒-8（CCK-8）检测CORT损伤的HT22细胞的活力，并用Hoechst 33258染色检测细胞凋亡。在动物实验中，将C57BL/6N小鼠随机分为对照组、模型组、低（100 mg·kg⁻¹）、中（200 mg·kg⁻¹）和高（400 mg·kg⁻¹）剂量的黄藤总生物碱组以及阳性对照组。经过21天的CUMS暴露后，通过行为学和Morris水迷宫试验观察其抑郁行为。采用酶联免疫吸附测定（ELISA）法评估血清5-羟色胺（5-HT）、多巴胺（DA）和去甲肾上腺素（NE）水平。通过蛋白质免疫印迹法检测HT22细胞和小鼠海马中凋亡相关蛋白Bcl-2、Bax和裂解的caspase-3的表达水平。结果表明，黄藤总生物碱以剂量依赖性方式减轻了CORT诱导的HT22细胞损伤。Hoechst 33258染色显示，黄藤总生物碱能更好地减少蓝色斑点并抑制细胞凋亡。动物实验结果表明，与模型组相比，黄藤总生物碱显著改善了小鼠的抑郁样行为，并提高了血清5-HT、DA和NE水平。蛋白质免疫印迹分析显示，黄藤总生物碱组中Bcl-2蛋白表达显著上调，而Bax和裂解的caspase-3蛋白表达明显降低。总之，黄藤总生物碱可能通过调节凋亡相关蛋白表达或提高脑中单胺类神经递质水平来抑制抑郁。
