摘要
目的探讨杜仲-续断药对治疗抑郁症的作用机制。方法通过中药系统网络药理学和分析平台(TCMSP)挖掘杜仲-续断药对的有效成分,通过GeneCards、OMIM和TTD数据库预测抑郁症疾病相关靶点,采用Venny 2.1.0软件获取交集靶点,采用String数据库和CytoScape软件构建蛋白-蛋白相互作用(PPI)网络,并进行基因本体论(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,采用AutoDock Tools 1.5.7软件对有效成分和节点度值排名靠前的关键靶点进行分子对接验证。将50只ICR小鼠随机分为对照组(等体积蒸馏水)、模型组(等体积蒸馏水)、杜仲-续断低剂量组(DXL,杜仲和续断各1.5 g/kg)、杜仲-续断高剂量组(DXH,杜仲和续断片各3 g/kg)和帕罗西汀组(20 mg/kg),各10只。采用慢性不可预知性刺激(CUMS)法复制抑郁症小鼠模型,建模成功后各组小鼠灌胃相应药物或蒸馏水,每天1次,连续18 d,期间采用CUMS法持续刺激。采用糖水偏好实验、悬尾实验、强迫游泳实验评估小鼠抑郁情况;采用酶联免疫吸附试验(ELISA)检测小鼠海马组织中多巴胺(DA)、5-羟色胺(5-HT)水平;采用RT-PCR法检测通路相关因子mRNA的表达水平。结果网络药理学分析结果显示,黄芩素、木犀草素、日本当药黄素、槲皮素、山柰酚、汉黄芩素为杜仲-续断药对治疗抑郁症的关键有效成分,蛋白激酶B1(AKT1)、雌激素受体1(ESR1)、磷脂酰肌醇-4,5-二磷酸3-激酶催化亚基α(PIK3CA)、原癌基因酪氨酸蛋白激酶Src(SRC)、表皮生长因子受体(EGFR)为重要潜在靶点;杜仲-续断药对治疗抑郁症的作用机制可能与磷脂酰肌醇3-激酶(PI3K)-AKT、钙信号、神经退行性病变等信号通路有关;分子对接结果显示,杜仲-续断药对主要有效成分与PIK3CA和AKT1结合稳定。动物实验结果显示,与模型组比较,DXL组、DXH组和帕罗西汀组小鼠糖水偏好率均显著升高(P<0.01),摇摆不动时间和漂浮不动时间(除DXL组外)均显著缩短(P<0.01),且DXH组和帕罗西汀组小鼠海马组织DA、5-HT水平均显著升高(P<0.05);DXL组小鼠海马组织AKT1 mRNA及DXH组小鼠海马组织PIK3CA和AKT1mRNA表达水平均显著升高(P<0.05)。结论杜仲-续断药对可有效改善模型小鼠抑郁症状,其机制可能与激活PI3K/AKT信号通路有关。
Objective To investigate the mechanism of Eucommiae Cortex-Dipsaci Radix herb pair in the treatment of depressive disorder.Methods TCMSP was used to mine the effective components of Eucommiae Cortex-Dipsaci Radix herb pair.The GeneCards,OMIM and TTD databases were used to predict the depressive disorder related targets.Venny 2.1.0 software was used to obtain intersection targets.String database and CytoScape software were used to construct the protein-protein interaction(PPI)network,and the gene ontology(GO)function enrichment analysis and the Kyoto Encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were carried out.AutoDock Tools 1.5.7 software was used to carry out the molecular docking validation of the top-ranked key targets of effective components and node degree value.A total of 50 ICR mice were randomly divided into control group(equal volume distilled water),model group(equal volume distilled water),Eucommiae Cortex-Dipsaci Radix herb pair low-dose group(DXL,1.5 g/kg for Eucommiae Cortex,Dipsaci Radix,respectively),Eucommiae Cortex-Dipsaci Radix herb pair high-dose group(DXH,3 g/kg for Eucommiae Cortex,Dipsaci Radix,respectively)and paroxetine group(20 mg/kg),with 10 mice in each group.Chronic unpredictable stimuli(CUMS)method was used to replicate the model of depressive disorder in mice.After successful modeling,the mice in each group were given corresponding drugs or distilled water by gavage administration once a day for 18 d,during which CUMS method was used for continuous stimulation.The sugar water preference test,tail suspension test and forced swimming test were used to evaluate the depressive disorder of mice;enzyme linked immunosorbent assay(ELISA)test was used to detect the levels of dopamine(DA)and 5-hydroxytryptamine(5-HT)in the hippocampus of mice;RT-PCR method was used to detect the mRNA expression level of pathway related factors.Results The results of network pharmacology analysis showed that baicalein,luteolin,swertiajaponin,quercetin,keampferol and wogonin were the key effective components of Eucommiae Cortex-Dipsaci Radix herb pair in the treatment of depressive disorder,and protein kinase B1(AKT1),estrogen receptor 1(ESR1),phosphatidylinositol-4,5-diphosphate 3-kinase catalytic subunitα(PIK3CA),proto oncogene tyrosine protein kinase SRC(SRC)and epidermal growth factor receptor(EGFR)were important potential targets;the mechanism of Eucommiae Cortex-Dipsaci Radix herb pair in the treatment of depressive disorder may be related to phosphatidylinositol 3-kinase(PI3K)-AKT,calcium signal,neurodegenerative lesions and other signaling pathways;the results of molecular docking showed that the main effective components of Eucommiae Cortex-Dipsaci Radix herb pair had stable binding with PIK3CA and AKT1.The results of animal experiments showed that compared with the model group,the sugar preference rate of DXL group,DXH group and paroxetine group were significantly increased(P<0.01),and the swinging immobility time and floating immobility time were significantly shortened in the DXH group and paroxetine group(P<0.01);the levels of DA and 5-HT in hippocampus of mice in the DXH group and paroxetine group were significantly increased(P<0.05);the expression mRNA level of AKT1 in hippocampus of mice in the DXL group,and the expression mRNA levels of PIK3CA and AKT1 in the hippocampus of mice in the DXH group were significantly increased(P<0.05).Conclusion Eucommiae Cortex-Dipsaci Radix herb pair can effectively improve the depressive disorder symptoms of model mice,and its mechanism may be related to the activation of PI3K/AKT signaling pathway.
作者
崔颖
杨磊
朱贲贲
王巍嵩
CUI Ying;YANG Lei;ZHU Benben;WANG Weisong(The Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010030,China;Community Health Service Center of Xincheng District East Street West Street,Hohhot,Inner Mongolia 010050,China;Peking University Cancer Hospital Inner Mongolia Hospital·Cancer Hospital Affiliated to Inner Mongolia Medical University,Hohhot,Inner Mongolia 010040,China)
出处
《中国药业》
2026年第8期44-51,共8页
China Pharmaceuticals
基金
内蒙古医科大学联合项目[YKD2022LH005]
内蒙古自治区公立医院科研联合基金科技项目[2023GLLH0123]。
关键词
杜仲-续断药对
抑郁症
网络药理学
分子对接
慢性不可预知性刺激
小鼠
Eucommiae Cortex-Dipsaci Radix pair
depressive disorder
network pharmacology
molecular docking
CUMS model mice
