摘要
目的:建立嗅球摘除(Olfactory Bulbectomy,OB)抑郁大鼠模型,从PI3K-AKT-Nrf2/BDNF角度探讨逍遥散抗抑郁作用机制。方法:手术摘除嗅球方法建立OB大鼠抑郁模型,将模型大鼠分为模型对照组、逍遥散15、30 g原生药/kg和盐酸氟西汀0.01 g/kg组,并设假手术组,各组大鼠连续灌胃相应药物或蒸馏水30天。进行大鼠旷场试验和糖水偏好率测定,腹主动脉取血分离血清,取大鼠皮质、海马部位备用。采用分光光度计法检测血清GSH、SOD及皮质MDA活力或含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测OB大鼠皮质和海马部位Nrf2、Keap1、GPX3、HO-1、NQO1、OGG1 mRNA的表达水平,采用蛋白免疫印迹法(Western blot)检测Nrf2、HO-1、SOD1、PI3K、AKT、p-AKT、TrkB、BDNF蛋白表达水平。结果:与假手术组比较,模型对照组大鼠血清GSH、SOD水平显著降低,且皮质MDA含量明显升高;皮质、海马部位Nrf2、Keap1、GPX3、HO-1 mRNA表达水平及SOD-1、HO-1、TrkB、BDNF蛋白表达水平显著下调,皮质部位PI3K蛋白表达和p-AKT/AKT比值显著降低。与模型对照组比较,15、30 g/kg逍遥散给药30 d能提高大鼠糖水偏好率,对抗自主活动异常,显著提高血清GSH、SOD水平,并降低皮质MDA含量;能显著上调皮质、海马部位Nrf2、HO-1、皮质Keap1、NQO1及海马GPX3 mRNA的表达水平,显著上调皮质、海马部位SOD1、HO-1、TrkB、BDNF蛋白表达水平及p-AKT/AKT比值,对皮质、海马部位OGG1mRNA及皮质Nrf2、海马PI3K的蛋白表达水平有上调趋势。结论:逍遥散对OB模型大鼠的行为学异常及氧化应激表现有对抗作用,其激活OB大鼠PI3K-AKT-Nrf2信号通路从而改善氧化应激状态、上调BDNF水平可能是其抗抑郁作用机制之一。
Objective:To establish a rat model with depression induced by Olfactory Bulbectomy(OB),and to study anti depression mechanism of Xiaoyao San by PI3 K-AKT-Nrf2/BDNF pass way.Methods:The OB-depressed rat model was established by the olfactory bulb surgically removed.All model rats were divided into the model group,Xiaoyao San groups(15 g、30 g raw drug/kg)and fluoxetine hydrochloride group(0.01 g/kg),in addition,the sham group was set.Rats were orally administered with corresponding drugs or saline water for 30 days.After administration,open field test and syrup preference test were performed.Blood was taken from the abdominal aorta and serum was collected.The cortex and hippocampus were dissected.Serum levels of GSH,SOD and cortical MDA were determined by spectrophotometry.The expressions of Nrf2,Keap1,GPX3,HO-1,NQO1 and OGG1 mRNA in cortex and hippocampus of OB rats were detected by real-time PCR.The expressions of Nrf2,HO-1,SOD1,PI3 K,AKT,p-AKT,TrkB and BDNF proteins in cortex and hippocampus of OB rats were detected by Western blot.Results:Compared with the sham group,serum levels of GSH and SOD in the model group were significantly decreased,the cortical MDA content was significantly increased.The expressions of Nrf2,Keap1,GPX3 and HO-1 mRNA and the expressions of SOD-1,HO-1,TrkB and BDNF proteins in the cortex and hippocampus of rats in the model group were significantly down-regulated,the PI3 K protein expression and p-AKT/AKT ratio were significantly reduced in the cortex.Compared with the model group,Xiaoyao san groups(15,30 g/kg)increased the sucrose preference rate of rats,reduced the abnormality of autonomic activity,increased the serum levels of GSH and SOD,and decreased the content of cortical MDA.It also regulated the expressions of Nrf2,HO-1,cortex Keap1,NQO1 and hippocampus GPX3 mRNA in cortex and hippocampus,and significantly up-regulated SOD1 and HO-1 in cortex and hippocampus and p-AKT/AKT ratio in the cortex.The cortex and hippocampus OGG1 mRNA,cortical Nrf2 and hippocampus PI3 K protein expressions were up-regulated.Conclusion:Xiaoyao san has an antagonistic effect on behavioral abnormalities and oxidative stress in OB model rats.It activates the PI3 K-AKT-Nrf2 signal pathway in OB rats.Its antidepressant mechanisms may be related to improving oxidative stress and up regulating BDNF level.
作者
罗杰
方洋
曾九僧
纪雅菲
刘蓉
曾南
Luo Jie;Fang Yang;Zeng Jiuseng;Ji Yafei;Liu Rong;Zeng Nan(Pharmacy College,Chengdu University of Traditional Chinese Medicine,Chengdu 611137)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2019年第6期2-6,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金(81503277)
四川省教育厅自然科学项目(16ZB0116)
成都中医药大学科研基金(ZRQN1643).
