柴胡皂苷A抗抑郁的靶点识别及作用研究  

Study on target identification and effect of saponin A on depressive disorder

作　　者：任历 邵钰婷 秦书华 杨文强 刘莹 REN Li;SHAO Yu-ting;QIN Shu-hua;YANG Wen-qiang;LIU Ying(College of Basic Medical Science,Jinzhou Medical University,Jinzhou 121000,China;Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500,China;Life Science of Institute Jinzhou Medical University,Jinzhou 121000,China)

机构地区：[1]锦州医科大学基础医学院,锦州121000 [2]昆明理工大学生命科学与技术学院,昆明650500 [3]锦州医科大学生命科学研究院,锦州121000

出　　处：《天然产物研究与开发》2024年第1期44-51,共8页Natural Product Research and Development

基　　金：辽宁省教育厅面上项目(LJKMZ20221248);国家自然科学基金地区基金(81560455)。

摘　　要：中药柴胡具有改善抑郁患者临床治疗效果的作用,柴胡皂苷A(saikosaponin A,SSA)是柴胡主要药效成分,本研究以SSA为研究对象,利用慢性不可预知性温和应激(chronic unpredictable mild stress,CUMS)模型小鼠悬尾(tail suspension test,TST)和强迫游泳实验(forced swimming test,FST)明确SSA的抗抑郁作用。利用计算机分子对接技术分析并验证SSA的作用靶点,并利用细胞热转移测定实验(cellular thermal shift assay,CETSA)和药物亲和力靶稳定性实验(drug affinity responsive target stability,DARTS)验证SSA的靶标;采用Western blotting等技术研究SSA的抗抑郁作用机制。CUMS模型TST和FST结果表明,与模型组相比,SSA能够显著缩短小鼠的不动时间,表明SSA具有显著的抗抑郁作用。计算机分子对接证明了SSA与催产素受体(oxytocin receptor,OXTR)结合效果较好,表明SSA抗抑郁的作用靶点可能是OXTR。CETSA结果表明在一系列温度梯度处理下,SSA能够明显延缓OXTR的热变性;DARTS实验结果表明,在一系列酶浓度梯度处理下,SSA能够显著降低OXTR对蛋白质水解的敏感性,表明SSA抗抑郁的作用靶点是OXTR。此外,SSA对OXTR下游的ERK通路没有激活作用。本研究表明SSA发挥抗抑郁作用的靶点可能为OXTR,为抑郁症等精神疾病的临床研究治疗和新药开发提供必要的理论与参考依据。Bupleuri Radix can improve the clinical treatment effect of depressed patients.Saikosaponin A(SSA)is the main medicinal component of Bupleurum Radix.Here,we take SSA as the research object,systematically identifies the anti-depression target of SSA and studies the anti-depression mechanism of SSA.The experiment of CUMS model mouse tail suspension test(TST)and forced swimming test(FST)were used to identify the anti-depression effect of SSA.The molecular docking experiment was used to analyze and verify the target of SSA.The target of SSA was further verified by cellular thermal shift assay(CETSA)and drug affinity responsive target stability(DARTS).In the mouse TST and FST,comparing the immobility time of the SSA experimental group and the model group,it was found that SSA can significantly shorten the immobility time of the mice,indicating that SSA has obviously antidepressant effects.Then using computer molecular docking,it was further found that the binding energy of SSA to the oxytocin receptor(OXTR)was low.The CETSA analysis showed that under a series of temperature gradient treatments,SSA can delay the thermal denaturation of OXTR.The DARTS analysis showed that under a series of enzyme concentration gradient treatments,SSA can reduce the sensitivity of OXTR to proteolysis.Western blotting analysis showed that SSA have no activating effect on the ERK pathway of OXTR.The study identified that the oxytocin receptor was the anti-depressant target of SSA,and preliminarily discussed the anti-depressant action mechanism of SSA,and provided the necessary theoretical basis in clinical treatment and new drug development.

关 键 词：抑郁症 柴胡皂苷A 催产素受体 细胞热转移测定实验 慢性不可预知性温和应激模型 

分 类 号：R93[医药卫生—生药学]