四逆散对抑郁大鼠BDNF/TrkB,5-HT/5-HT1AR及HPA轴的影响  

Effect of Sinisan on BDNF/TrkB,5-HT/5-HT1AR,and HPA axis in Depression Model Rats

作　　者：李耀洋 尚立芝 毛梦迪[2,3] 王伟杰 陈晓辉[3] 周春雨[2,3] 张光远 闫鸿豪 LI Yao-yang;SHANG Li-zhi;MAO Meng-di;WANG Wei-jie;CHEN Xiao-hui;ZHOU Chun-yu;ZHANG Guang-yuan;YAN Hong-hao(The First Clinical School of Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Key Laboratory for Modern Research of Zhongjing Compound Traditional Chinese Medicine,Zhengzhou 450046,China;Academy of Chinese Medical Sciences,Henan University of Chinese Medicine,Zhengzhou 450046,China;First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450006,China)

机构地区：[1]河南中医药大学第一临床医学院,郑州450046 [2]河南省仲景方药现代研究重点实验室,郑州450046 [3]河南中医药大学中医药科学院,郑州450046 [4]河南中医药大学第一附属医院,郑州450006

出　　处：《中国实验方剂学杂志》2021年第24期40-48,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省高等学校重点科研项目(20B360011,21A630020);国家级大学生创新创业训练计划支持项目(202010471016);河南省大学生创新创业训练计划支持项目(S202010471003);河南中医药大学大学生创新学习项目(CXXM2020016,CXXM2020003)。

摘　　要：目的:观察四逆散对抑郁大鼠脑神经营养因子(BDNF)/酪氨酸激酶受体B(TrkB),5-羟色胺(5-HT)/5-HT1A受体(5-HT1AR)及下丘脑-垂体-肾上腺(HPA)轴的影响,探讨四逆散基于BDNF/TrKB,5-HT/5-HT1AR及HPA轴的抗抑郁作用机制。方法:120只雄性Wistar大鼠随机分为正常组、模型组、四逆散低、中、高剂量组和氟西汀组,每组20只。除正常组外,其余5组连续21 d采用孤养结合慢性温和不可预知性刺激法(CUMS)制备抑郁大鼠模型。四逆散低、中、高剂量组分别灌胃(ig)四逆散(1.25,2.5,5)g·kg^(-1),氟西汀组ig盐酸氟西汀0.01 g·kg^(-1),正常组和模型组ig等体积生理盐水,每日1次,持续干预21 d。干预期间,各造模组大鼠均继续给予刺激。通过糖水偏好和旷场实验评估CUMS模型大鼠抑郁状态;酶联免疫吸附测定法(ELISA)检测血浆促肾上腺皮质激素释放激素(CRH),促肾上腺皮质激素(ACTH),皮质酮(CORT)及海马匀浆中BDNF,5-HT水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测海马TrKB,5-HT1AR,糖皮质激素受体(GR)和盐皮质激素受体(MR)mRNA表达;蛋白免疫印迹法(Western blot)检测海马TrKB,5-HT1AR,GR,MR蛋白表达水平;苏木素-伊红(HE)染色观察海马组织形态学变化。结果:与正常组比较,模型组大鼠糖水偏好率显著下降(P<0.01),旷场实验中水平得分和垂直得分显著降低(P<0.01),血浆CRH,ACTH,CORT含量显著升高(P<0.01),海马BDNF,5-HT含量显著降低(P<0.01),海马TrKB,5-HT1AR和GR mRNA及蛋白表达水平显著降低(P<0.01),MR mRNA及蛋白表达水平显著升高(P<0.01),HE染色结果显示海马神经元结构损伤。与模型组比较,四逆散低、中、高剂量组和氟西汀组糖水偏好率显著升高(P<0.01),旷场实验中水平得分和垂直得分明显升高(P<0.05,P<0.01),血浆CRH,ACTH,CORT含量明显降低(P<0.05,P<0.01),海马BDNF,5-HT含量明显升高(P<0.05,P<0.01),海马TrKB,5-HT1AR和GR mRNA及蛋白表达水平明显升高(P<0.05,P<0.01),MR mRNObjective:To observe the effect of Sinisan on the brain-derived neurotrophic factor(BDNF)/tyrosine kinase receptor B(TrKB),5-hydroxytryptamine(5-HT)/5-HT1A receptor(5-HT1AR),and hypothalamus-pituitary-adrenal(HPA)axis in depressed rats,and explore the antidepressant mechanism of Sinisan based on BDNF/TrKB,5-HT/5-HT1AR,and HPA axis.Method:A total of 120 male Wistar rats were randomly divided into a normal group,a model group,a fluoxetine(0.01 g·kg^(-1)) group,and low-(1.25 g·kg^(-1)),medium-(2.5 g·kg^(-1)),and high-dose(5 g·kg^(-1))Sinisan groups,with 20 rats in each group.The depression model was induced by isolation combined with chronic unpredictable mild stimulation(CUMS)in rats except for those in the normal group for 21 days.Rats were then treated correspondingly once a day for 21 days by gavage.Those in the normal group and the model group received an equal volume of normal saline.During the intervention,the model rats were stimulated continuously.The depressive state of CUMS model rats was evaluated by sucrose preference test and open field test.Enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),and corticosterone(CORT)in the plasma and BDNF and 5-HT levels in the hippocampal homogenate.The mRNA expression of hippocampal TrKB,5-HT1AR,glucocorticoid receptor(GR),and mineralocorticoid receptor(MR)was detected by real-time fluorescence-based quantitative polymerase chain reaction(Real-time PCR).The protein expression of hippocampal TrKB,5-HT1AR,GR,and MR was detected by Western blot.The histomorphological changes of the hippocampus were observed by hematoxylin-eosin(HE) staining.Result:Compared with the normal group,the model group showed decreased sucrose preference rate(P<0.01),reduced horizontal and vertical scores in the open field test(P<0.01),increased plasma content of CRH,ACTH,and CORT(P<0.01),declining content of BDNF and 5-HT in the hippocampus(P<0.01),dwindled mRNA and protein expression levels of Tr

关 键 词：四逆散 抑郁症 脑源性神经营养因子(BDNF) 酪氨酸激酶受体B(TrKB) 下丘脑-垂体-肾上腺(HPA)轴 糖皮质激素受体(GR) 盐皮质激素受体(MR) 

分 类 号：R2-0[医药卫生—中医学] R22