柴胡疏肝散调节围绝经期肝郁证大鼠ERK1/2-CREB-BDNF信号通路的机制  

Mechanism of Chaihu Shugan Powder on ERK1/2-CREB-BDNF signaling pathway in perimenopausal liver-stagnation pattern rats

作　　者：梁文娜[1,2,3] 阮璐薇 张鹏横 沈建英 黄睿婷[1,2,3] LIANG Wen-na;RUAN Lu-wei;ZHANG Peng-heng;SHEN Jian-ying;HUANG Rui-ting(Chinese Medicine School,Fujian University of Traditional Chinese Medicine,Fuzhou 35122,China;Fujian Province Collaborative Innovation Center of TCM Health Management in 211,Fuzhou 35122,China;Key Laboratory of TCM Health Status Identification in Fujian Province,Fuzhou 35122,China)

机构地区：[1]福建中医药大学中医学院,福州35122 [2]福建省2011中医健康管理协同创新中心,福州35122 [3]福建省中医健康状态辨识重点实验室,福州35122

出　　处：《中华中医药杂志》2022年第6期3147-3152,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81774209);福建省科技厅社会发展引导性项目(No.2019Y0039);福建省自然科学基金项目(No.2018J01894);全国中医药创新骨干人才培训项目(No.国中医药人教函[2019]128号)。

摘　　要：目的:探讨柴胡疏肝散调节围绝经期肝郁证大鼠ERK1/2-CREB-BDNF信号通路的作用及机制。方法:采用去势联合慢性不可预知性温和应激(CUMS)建立围绝经期肝郁证大鼠模型,按体质量分层随机分为假手术组(0.9%氯化钠溶液)、围绝组(去势+0.9%氯化钠溶液)、围绝肝郁组(去势+CUMS+0.9%氯化钠溶液)和柴胡疏肝散组(去势+CUMS+柴胡疏肝散),每天1次,连续干预21 d。检测强迫游泳不动时间、1%蔗糖水偏好率的行为学变化,ELISA法检测血清E、ACTH、CORT、5-HT、DA、NE含量变化。Real-time PCR检测海马ERK1/2、CREB、BDNF、Trk B mRNA表达,Western Blot检测海马ERK1/2、CREB、BDNF、TrkB蛋白表达。结果:与假手术组比较,围绝肝郁组强迫游泳不动时间显著延长(P<0.05),1%蔗糖水偏好率显著降低(P<0.01);与围绝组比较,围绝肝郁组1%蔗糖水偏好率显著降低(P<0.01);与围绝肝郁组比较,柴胡疏肝散组强迫游泳不动时间显著缩短(P<0.01),1%蔗糖水偏好率显著升高(P<0.01)。与假手术组比较,各组血清E含量显著降低(P<0.01)。与假手术组和围绝组比较,围绝肝郁组血清ACTH、CORT含量均显著升高(P<0.01),5-HT、DA、NE含量显著降低(P<0.01),而柴胡疏肝散组较围绝肝郁组血清ACTH、CORT含量显著降低(P<0.01),5-HT、DA、NE含量显著升高(P<0.01)。与假手术组比较,围绝肝郁组海马ERK1/2、BDNF、TrkB mRNA表达量显著降低(P<0.05,P<0.01),与围绝组比较,围绝肝郁组海马ERK1/2、GREB、BDNF mRNA表达显著降低(P<0.01),而柴胡疏肝散组较围绝肝郁组海马ERK1/2、CREB、BDNF、TrkB mRNA表达显著升高(P<0.01)。与假手术组比较,围绝肝郁组海马BDNF、TrkB蛋白水平显著降低(P<0.01,P<0.05);与围绝组比较,围绝肝郁组海马CREB蛋白水平显著下降(P<0.05);而柴胡疏肝散组较围绝肝郁组海马CREB、BDNF、TrkB蛋白水平显著升高(P<0.05,P<0.01)。结论:柴胡疏肝散通过调节神经递质5-HT、DA、NE表达紊乱与ERK1/2-CREB-Objective:To explore mechanism of Chaihu Shugan Powder on ERK1/2-CREB-BDNF signaling pathway in perimenopausal liver-stagnation pattern rats.Methods:Female 2-month-old SPF-grade SD rats were established the animal model of perimenopausal liver-stagnation pattern using ovariectomy combined with chronic unpredictable mild stress(CUMS),and randomly divided into sham group(saline),perimenopause(PMS)group(ovariectomy+saline),perimenopausal liver-stagnation pattern(PLSP)group(ovariectomy+CUMS+saline)and Chaihu Shugan Powder(CSS)group(ovariectomy+CUMS+Chaihu Shugan Powder)according to body weight of rats,once a day for 21 consecutive days.The changes of immobility time of forced swimming and 1%sucrose preference rate were tested.Serum E,ACTH,CORT,5-HT,DA and NE content was detected by enzyme-linked immunosorbent assay(ELISA).mRNA and ptotein expressions of ERK1/2,CREB,BDNF and TrkB in hippocampus were measured by real-time quantitative PCR(q-PCR)and Western Blot,respectively.Results:Compared with the sham group,the immobility time of forced swimming in the PLSP group was prolonged(P<0.05),and the 1%sucrose preference rate was significantly reduced in the PLSP group(P<0.01).Compared with the PMS group,the 1%sucrose water preference rate was significantly reduced in the PLSP group(P<0.01).Compared with the PLSP group,the immobility time of forced swimming in the CSS group was shorter(P<0.01),the 1%sucrose water preference rate in the CSS group was significantly increased(P<0.01).Compared with the sham group,the serum Econtent in rats was significantly reduced in each group(P<0.01).The serum ACTH and CORT contents in the PLSP group significantly increased than that in the sham and PMS group(P<0.01),while the CSS group significantly decreased than the PLSP group(P<0.01).The contents of serum 5-HT,DA,and NE in the PLSP group significantly were down-regulated than that in the sham and PMS group(P<0.01),and the CSS group was significantly up-regulated than the PLSP group(P<0.01).Hippocampal ERK1/2,BDNF and TrkB mRNA expression

关 键 词：围绝经期 肝郁证 ERK1/2-CREB-BDNF信号通路 柴胡疏肝散 机制 神经递质 动物模型 行为学变化 

分 类 号：R285.5[医药卫生—中药学]