摘要
目的探究芒柄花素(FMN)对抑郁大鼠的改善作用及分子机制。方法将72只SD大鼠分为对照组、模型组、氟西汀组(2 mg/kg)、FMN低剂量组(30 mg/kg)、FMN高剂量组(60 mg/kg)、FMN+ML385(核因子E2相关因子2抑制剂)组(FMN 60 mg/kg+ML38530 mg/kg),每组12只。除对照组外,其余各组大鼠采用孤养结合慢性不可预知性温和应激(CUMS)诱导建立抑郁症模型。给予相应的干预后,糖水偏好实验(SPT)和强迫游泳实验(FST)检测大鼠的行为学变化,试剂盒检测血清促肾上腺皮质激素(ACTH)、皮质酮(CORT)水平和海马组织中丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性,Nissl染色和TUNEL染色观察大鼠海马神经元损伤情况,Western blot检测海马组织核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路相关蛋白[Nrf2、醌氧化还原酶1(NQO1)、血红素氧合酶-1(HO-1)]表达。结果与对照组相比,模型组大鼠SPT中糖水偏好系数、海马组织SOD和CAT活性、Nissl阳性细胞数、细胞质和核内Nrf2蛋白水平以及NQO-1、HO-1蛋白水平显著降低,差异有统计学意义(P均<0.05),FST中的不动时间、血清ACTH、CORT水平、海马组织MDA含量、海马神经元凋亡率显著升高,差异有统计学意义(P均<0.05);与模型组相比,氟西汀组和FMN低、高剂量组大鼠SPT中糖水偏好系数、海马组织SOD和CAT活性、Nissl阳性细胞数、细胞质和核内Nrf2蛋白水平以及NQO-1、HO-1蛋白水平显著升高,差异有统计学意义(P均<0.05),FST中的不动时间、血清ACTH、CORT水平、MDA含量、海马神经元凋亡率显著降低,差异有统计学意义(P<0.05);氟西汀组与FMN高剂量组间以上项目比较差异无统计学意义(P>0.05)。与FMN高剂量组相比,FMN+ML385组大鼠SPT中糖水偏好系数、海马组织SOD和CAT活性、Nissl阳性细胞数、细胞质和核内Nrf2蛋白水平以及NQO-1、HO-1蛋白水平显著降低,FST中的不动时间、血清ACTH、CORT水平、海马组织MDA含量、海马神经元凋亡率显著增加,差异均有统计学意义(P均<0.05)。结论FMN可抑制氧化应激,减轻海马神经元损伤,改善CUMS诱导的抑郁大鼠的抑郁样行为,其作用机制可能与激活Nrf2/ARE信号通路有关。
Objective To explore the improvement effect and molecular mechanism of formononetin(FMN)on depressedrats.MethodsSeventy-two SD rats were grouped into control group,model group,fluoxetine group(2 mg/kg),FMN low-dose group(30 mg/kg),FMN high-dose group(60 mg/kg),and FMN+ML385 group(FMN 60 mg/kg+ML38530 mg/kg),with 12 animals in each group.Except for the control group,rats in other groups were induced to establish depressionmodels by solitary feeding combined with chronic unpredictable mild stress(CUMS).After corresponding interventions,sugar water preference test(SPT)and forced swimming test(FST)were used to detect the behavioral changes of rats;thekits were used to detect serum levels of adrenocorticotropic hormone(ACTH),corticosterone(CORT),malondialdehyde(MDA)content,superoxide dismutase(SOD),and catalase(CAT)activities in hippocampus tissue;Nissl staining andTUNEL staining were used to observe the damage of rat hippocampal neurons;Western blot was used to measure theexpression of nuclear factor erythroid-2-related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway-related proteins[Nrf2,quinone oxidoreductase 1(NQO1),heme oxygenase-1(HO-1)]in hippocampus.ResultsComparedwith the control group,the sucrose preference coefficient in SPT,activities of SOD and CAT in hippocampus,the numberof Nissl positive cells,the protein level of Nrf2 in cytoplasm and nucleus,and the protein levels of NQO-1 and HO-1 in themodel group were obviously decreased(P<0.05);the immobility time in FST,the levels of serum ACTH and CORT,thecontent of hippocampal MDA,and hippocampal neuron apoptosis rate were obviously increased,with statisticallysignificance(P<0.05);compared with the model group,the sucrose preference coefficient in SPT,activities of SOD andCAT in hippocampus,the number of Nissl positive cells,the protein level of Nrf2 in cytoplasm and nucleus,and theprotein levels of NQO-1 and HO-1 in the fluoxetine group and the FMN low and high-dose groups were obviously increased(P<0.05);the immobility time in FST,the levels of serum ACTH and CORT,the content of hippocampal MDA,andhippocampal neuron apoptosis rate were obviously decreased,with statistically significance(P<0.05);there was no obviousdifference between the fluoxetine group and the FMN high-dose group(P>0.05).Compared with the FMN high-dose group,the sugar water preference coefficient,the activity of SOD and CAT in hippocampus,the number of Nissl positive cells,theprotein levels of Nrf2 in cytoplasm and nucleus,and the protein levels of NQO-1 and HO-1 were significantly decreased inFMN+ML385 group.The immovable time,serum ACTH and CORT levels,MDA content in hippocampal tissue and apoptosisrate of hippocampal neurons in FST were significantly increased,with statistically significance(P<0.05).ConclusionFMNcould inhibit oxidative stress,reduce hippocampal neuron damage,and improve depression-like behavior in CUMS-induceddepression rats,and its mechanism might be related to the activation of Nrf2/ARE signaling pathway.
作者
程瑶
翟冠楠
王春霞
刘强
CHENG Yao;ZHAI Guan-nan;WANG Chun-xia;LIU Qiang(Department of Encephalopathy,Tai'an Traditional Chinese Medicine Hospital,Tai'an,Shandong 271000,China;Health Management Center,Tai'an Traditional Chinese Medicine Hospital,Tai'an,Shandong 271000,China)
出处
《热带医学杂志》
CAS
2022年第11期1467-1472,1486,1611,共8页
Journal of Tropical Medicine
基金
山东省中医药科技项目(2021M063)
作者简介
Corresponding author:程瑶(1982-),女,硕士,副主任医师,主要从事脑病,卒中后抑郁方面研究,E-mail:cy271000@163.com
