文献类型：期刊文献

中文题名：基于HPA轴-MEK/ERK信号通路探讨柴胡加龙骨牡蛎汤治疗广泛性焦虑症大鼠的作用机制

英文题名：Exploration on the Mechanism of Chaihu Longgu Muli Decoction in the Treatment of Generalized Anxiety Disorder Rats Based on the HPA Axis-MEK/ERK Signaling Pathway

作者：胡亦明[1,2];杨蕤[1,2];韦春昕[1,2];黄仕琦[1,2];谭国伟[1,2];李晓[1];安耀荣[1]

第一作者：胡亦明

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃省中医方药挖掘与创新转化重点实验室,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2025

卷号：32

期号：6

起止页码：105

中文期刊名：中国中医药信息杂志

外文期刊名：Chinese Journal of Information on Traditional Chinese Medicine

基金：甘肃省自然科学基金(22JR11RA121);甘肃中医药大学中医学一级学科“岐黄英才”导师专项基金硕士研究生导师开放基金(3016190124)。

语种：中文

中文关键词：广泛性焦虑症;柴胡加龙骨牡蛎汤;下丘脑-垂体-肾上腺轴;MEK/ERK信号通路;氧化应激;大鼠

外文关键词：generalized anxiety disorder;Chaihu Longgu Muli Decoction;hypothalamic-pituitary-adrenal axis;MEK/ERK signaling pathway;oxidative stress;rats

摘要：目的观察柴胡加龙骨牡蛎汤对广泛性焦虑症(GAD)大鼠海马组织氧化应激和下丘脑-垂体-肾上腺(HPA)轴激素水平的影响,探讨其缓解GAD大鼠焦虑的作用机制。方法72只Wistar大鼠随机分为空白组12只和造模组60只,采用慢性束缚应激法进行GAD造模。将造模成功大鼠随机分为模型组、地西泮组(1 mg/kg)和中药低、中、高剂量组(柴胡加龙骨牡蛎汤6、12、24 g/kg),给药组分别予相应药液灌胃,连续14 d。旷场实验、高架十字迷宫实验观察大鼠行为学指标,HE染色观察海马组织形态,ELISA检测血清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)含量,生化试剂盒检测海马组织丙二醛(MDA)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)含量,RT-PCR检测海马组织MEK、ERK、Bax、Bcl-2 mRNA表达,Western blot检测海马组织MEK、p-MEK、ERK、p-ERK、Bax、Bcl-2蛋白表达。结果与空白组比较,模型组大鼠体质量增长缓慢,毛色晦黯、脱毛明显,表现为极度焦躁不安;中央区域活动时间和活动距离显著减少(P<0.01),进入开放臂时间、次数和活动距离百分比显著减少(P<0.01);海马组织神经元结构变性,细胞形态改变,排列紊乱,周围组织水肿;血清CRH、ACTH、CORT含量显著升高(P<0.01);海马组织MDA含量显著升高(P<0.01),CAT、SOD含量显著降低(P<0.01),p-MEK、p-ERK蛋白表达显著降低(P<0.01),Bax mRNA和蛋白表达显著升高(P<0.01),Bcl-2 mRNA和蛋白表达显著降低(P<0.01)。与模型组比较,地西泮组和中药各剂量组大鼠体质量增长速度提高,脱毛、焦躁、抱团蜷缩有所改善;地西泮组和中药中、高剂量组大鼠中央区域活动时间和活动距离显著增加(P<0.01),进入开放臂时间、次数和活动距离百分比显著增加(P<0.05,P<0.01);血清CRH、ACTH、CORT含量显著降低(P<0.05,P<0.01);地西泮组和中药高剂量组大鼠海马组织MDA含量显著降低(P<0.01),CAT、SOD含量显著升高(P<0.05,P<0.01),地西泮组和中药中、高剂量组大鼠海马组织p-MEK、p-ERK蛋白表达显著降低(P<0.01),Bax mRNA和蛋白表达显著降低(P<0.01),Bcl-2 mRNA和蛋白表达显著升高(P<0.05,P<0.01)。各组MEK、ERK mRNA及蛋白表达差异无统计学意义(P>0.05)。结论柴胡加龙骨牡蛎汤能缓解GAD大鼠焦虑状态,其机制与抑制HPA轴激素释放和MEK/ERK信号通路活化,改善机体神经内分泌水平,降低海马氧化应激反应有关。
Objective To observe the effects of Chaihu Longgu Muli Decoction on oxidative stress in hippocampus and hypothalamic-pituitary-adrenal(HPA)axis hormone levels of rats with generalized anxiety disorder(GAD);To explore its mechanism of alleviating anxiety in GAD rats.Methods A total of 72 Wistar rats were randomly divided into blank group(12 rats)and modeling group(60 rats).Chronic restraint stress was used to establish the GAD model.The successfully modeled rats were randomly divided into model group,diazepam group(1 mg/kg)and TCM low-,medium-and high-dosage groups(Chaihu Longgu Muli Decoction 6,12,24 g/kg).The administration groups were given corresponding drugs by gavage for 14 consecutive days.Open field experiment and elevated cross maze experiment were used to observe indexes in rat behavior,HE staining was used to observe the morphology of hippocampal tissue,ELISA was used to detect the contents of serum corticotropin releasing hormone(CRH),adrenocorticotropic hormone(ACTH)and corticosterone(CORT),biochemical kits were used to detect the contents of malondialdehyde(MDA),catalase(CAT)and superoxide dismutase(SOD)in hippocampal tissue,RT-PCR was used to detect the mRNA expressions of MEK,ERK,Bax and Bcl-2 in hippocampal tissue,Western blot was used to detect the protein expressions of MEK,p-MEK,ERK,p-ERK,Bax and Bcl-2 in hippocampal tissue.Results Compared with the blank group,the rats in the model group had a significantly slower growth rate of body msss,loose stool,dark hair,significant hair removal and extreme agitation;the activity time and distance in the central area were significantly reduced(P<0.01),the percentage of time,frequency and distance of entering the open arm were significantly reduced(P<0.01);with structural degeneration of hippocampal neurons,changes in cell morphology,disordered arrangement and edema of surrounding tissues;the contents of serum CRH,ACTH and CORT significantly increased(P<0.01);the MDA content in hippocampal tissue significantly increased(P<0.01),CAT and SOD contents significantly decreased(P<0.01),p-MEK and p-ERK protein expressions significantly decreased(P<0.01),Bax mRNA and protein expressions significantly increased(P<0.01),Bcl-2 mRNA and protein expressions significantly decreased(P<0.01).Compared with the model group,the body mass growth rate of diazepam group and each dosage of TCM group increased,and the symptoms of loose stool,hair removal,anxiety and huddling were improved;the activity time and distance in the central area of rats in the diazepam group and TCM medium-and high-dosage groups significantly increased(P<0.01),and the percentage of time,frequency and distance of entering the open arm significantly increased(P<0.05,P<0.01);the contents of serum CRH,ACTH and CORT significantly decreased(P<0.05,P<0.01);the MDA content in the hippocampal tissue in diazepam group and TCM high-dosage group significantly decreased(P<0.01),while the CAT and SOD contents significantly increased(P<0.05,P<0.01),the protein expressions of p-MEK and p-ERK in hippocampal tissue in diazepam group and TCM medium-and high-dosage group significantly decreased(P<0.01),Bax mRNA and protein expressions were significantly decreased(P<0.01),and Bcl-2 mRNA and protein expressions significantly increased(P<0.05,P<0.01).There was no statistical significance in the expressions of MEK,ERK mRNA and protein among the groups(P>0.05).Conclusion Chaihu Longgu Muli Decoction can alleviate anxiety in GAD rats by inhibiting HPA axis hormone release and MEK/ERK signaling pathway activation,improving neuroendocrine levels,and reducing hippocampal oxidative stress response.