[关键词]
[摘要]
目的 通过皮质酮诱导的小鼠抑郁症模型和肾上腺嗜铬细胞瘤PC-12细胞损伤模型,研究怀菊花提取物的抗抑郁作用。方法 将雄性C57BL/6小鼠随机分为对照组、模型组、氟西汀(5 mg/kg)组和怀菊花提取物低、中、高剂量(1.67、3.33、6.67 g/kg)组,模型组和各给药组连续21 d sc皮质酮(20 mg/kg),造模同时ig相应药物,给药结束后进行糖水偏好实验、悬尾实验和旷场实验;采用ELISA法检测小鼠血清中皮质酮及神经递质水平;采用Western blotting检测海马组织中环磷腺苷效应元件结合蛋白(cyclic adenosine monophosphate-response element binding protein,CREB)通路关键蛋白表达。皮质酮诱导PC-12细胞建立细胞损伤模型,采用流式细胞术检测怀菊花提取物对细胞凋亡及活性氧(reactive oxygen species,ROS)水平的影响;采用In-Cell Western法检测凋亡相关蛋白及CREB通路关键蛋白表达。结果 怀菊花提取物能够显著改善小鼠的糖水偏好率(P<0.05、0.01),增加小鼠的站立次数(P<0.05、0.01),减少小鼠悬尾实验的不动时间(P<0.01),显著降低血清中皮质酮水平(P<0.01),升高血清中5-羟色胺(5-hydroxytryptamine,5-HT)、去甲肾上腺素(norepinephrine,NE)、多巴胺(dopamine,DA)、乙酰胆碱(acetylcholine,Ach)水平(P<0.05、0.01),上调海马组织中CREB、磷酸化CREB(phosphorylated CREB,p-CREB)、环磷酸腺苷(cyclic adenosine monophosphate,cAMP)蛋白表达水平(P<0.05、0.01)。怀菊花提取物显著抑制皮质酮诱导的PC-12细胞凋亡率及细胞内ROS水平(P<0.01),抑制凋亡相关蛋白表达(P<0.01),上调CREB通路关键蛋白表达(P<0.01)。结论 怀菊花提取物可能通过调节CREB通路、抑制神经细胞凋亡,从而改善抑郁。
[Key word]
[Abstract]
Objective To study antidepressant effect of Huai Juhua (Chrysanthemi Flos) extract on corticosterone-induced mouse depression model and adrenal pheochromocytoma PC-12 cell injury model. Methods Male C57BL/6 mice were randomly divided into control group, model group, fluoxetine (5 mg/kg) group and Huai Chrysanthemi Flos extract low-, medium-and high-dose (1.67, 3.33, 6.67 g/kg) groups. Model group and each administration group were sc corticosterone (20 mg/kg) for 21 consecutive days, and corresponding drugs were given at the same time during modeling. After administration, sugar water preference experiment, tail suspension experiment and open field experiment were performed; Corticosterone and neurotransmitter levels in serum of mice were detected by kits; Western blotting was used to detect the expression of key proteins in cyclic adenosine monophosphate-response element binding protein (CREB) pathway in hippocampus. Corticosterone induced PC-12 cells was used to establish a cell injury model, flow cytometry was used to detect the effect of Huai Chrysanthemi Flos extract on cell apoptosis and reactive oxygen species (ROS) levels; In-Cell Western method was used to detect apoptosis-related protein and CREB pathway key protein expressions. Results Huai Chrysanthemi Flos extract significantly improved the sugar water preference rate of mice (P < 0.05, 0.01), increased the number of standing times (P < 0.05, 0.01), reduced the immobility time of mice in tail suspension test (P < 0.01), significantly decreased corticosterone level in serum (P < 0.01), increased levels of 5-hydroxytryptamine (5-HT), norepinephrine (NE), dopamine (DA) and acetylcholine (Ach) in serum (P < 0.05, 0.01), up-regulated the protein expression levels of CREB, phosphorylated CREB (p-CREB) and cyclic adenosine monophosphate (cAMP) in hippocampus (P < 0.05, 0.01). Huai Chrysanthemi Flos extract significantly inhibited corticosterone-induced PC-12 cells apoptosis and intracellular ROS level (P < 0.01), inhibited the expression of apoptosis-related proteins (P < 0.01), and up-regulated the expression of key proteins in CREB pathway (P < 0.01). Conclusion Huai Chrysanthemi Flos extract may improve depression by regulating CREB pathway and inhibiting neuronal apoptosis.
[中图分类号]
R285.5
[基金项目]
国家重点研发计划项目(2017YFC1702800);河南省中医药科学研究专项课题(20-21ZY2148);河南省重大科技专项(171100310500);河南省高层次人才特殊支持“中原千人计划”项目(ZYQR201810080);河南省教育厅河南省科技攻关项目(212102311106);河南中医药大学2018年度博士科研基金资助项目(BSJJ2018-04)