木兰花碱减轻慢性应激小鼠抑郁样行为及对脑部LSD1组蛋白修饰的影响
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摘要
目的探索木兰花碱对慢性不可预见性温和应激所致抑郁小鼠行为学的影响机制。方法将ICR小鼠随机分为正常对照组(control组)、抑郁模型组(PG组)、木兰花碱高剂量组(MH组)和木兰花碱低剂量组(ML组)。采用慢性不可预见性温和刺激方法建立抑郁小鼠模型,检测各组小鼠行为学的变化,并用实时定量PCR、Western blot检测小鼠脑部赖氨酸特异性组蛋白去甲基化酶1(LSD1)mRNA及蛋白表达水平的变化。免疫组化染色法检测小鼠脑部LSD1阳性细胞数量。结果木兰花碱在行为学上改善了小鼠的抑郁状况,与PG组相比,MH组和ML组悬尾不动时间、强迫游泳不动时间明显缩短(P<0.05)。实时定量PCR及Western blot结果显示,与control组相比,PG组LSD1mRNA表达差异无统计学意义,蛋白表达明显降低(P<0.05),ML组LSD1 mRNA和蛋白表达明显升高(P<0.05);与PG组相比,ML组LSD1 mRNA和蛋白表达明显升高(P<0.05)。免疫组化结果显示,与control组相比,PG组小鼠脑部LSD1阳性细胞数量减少,而ML组较PG组增加。结论在木兰花碱治疗抑郁小鼠的过程中,LSD1可能起到了重要的调节作用。
Objective To explore the mechanism of the effect of magnoflorine on the behavior of mice with depression caused by chronic unpredictable mild stress.MethodsThe ICR mice were randomly divided into normal control group(control group), positive depression group(PG group), high dose of magnoflorine group(MH group) and low dose of magnoflorine group(ML group). Depression mouse model was established by chronic unpredictable mild stress method. The behavioral changes of mice were detected. The expressions of lysine specific demethylase 1(LSD1) mRNA and protein in brain of mice were detected by real-time quantitative PCR, Western blot assay and immunohistochemistry.Results Magnoflorine improved the depression in mice. Compared with the PG group, the motionless time of tail suspension and motionless time of forced swimming were significantly shorter in the MH group and the ML group(P<0.05). Real-time quantitative PCR and Western blot assay showed that there was no significant difference in LSD1 mRNA expression between the PG group and the control group. The protein expression was significantly decreased(P<0.05). The expressions of LSD1 mRNA and protein were significantly increased in the ML group compared with those of control group(P<0.05). Compared with the PG group, the expressions of LSD1 mRNA and protein were significantly increased in the ML group(P<0.05). The results of immunohistochemistry showed that the number of positive cells in brain was decreased in the PG group compared with that of the control group, while it was increased in the ML group compared with that of PG group.ConclusionLSD1 may play an important role in the treatment with magnoflorine for depressed mice.
Objective To explore the mechanism of the effect of magnoflorine on the behavior of mice with depression caused by chronic unpredictable mild stress.MethodsThe ICR mice were randomly divided into normal control group(control group), positive depression group(PG group), high dose of magnoflorine group(MH group) and low dose of magnoflorine group(ML group). Depression mouse model was established by chronic unpredictable mild stress method. The behavioral changes of mice were detected. The expressions of lysine specific demethylase 1(LSD1) mRNA and protein in brain of mice were detected by real-time quantitative PCR, Western blot assay and immunohistochemistry.Results Magnoflorine improved the depression in mice. Compared with the PG group, the motionless time of tail suspension and motionless time of forced swimming were significantly shorter in the MH group and the ML group(P<0.05). Real-time quantitative PCR and Western blot assay showed that there was no significant difference in LSD1 mRNA expression between the PG group and the control group. The protein expression was significantly decreased(P<0.05). The expressions of LSD1 mRNA and protein were significantly increased in the ML group compared with those of control group(P<0.05). Compared with the PG group, the expressions of LSD1 mRNA and protein were significantly increased in the ML group(P<0.05). The results of immunohistochemistry showed that the number of positive cells in brain was decreased in the PG group compared with that of the control group, while it was increased in the ML group compared with that of PG group.ConclusionLSD1 may play an important role in the treatment with magnoflorine for depressed mice.
引文
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[17]Wang P,Fan F,Li X,et al.Riboflavin attenuates myocardial injury via LSD1-mediated crosstalk between phospholipid metabolism and histone methylation in mice with experimental myocardial infarction[J].J Mol Cell Cardiol,2018,115:115-129.doi:10.1016/j.yjmcc.2018.01.006.
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[20]Teply RM,Packard KA,White ND,et al.Treatment of depression in patients with concomitant cardiac disease[J].Prog Cardiovasc Dis,2016,58(5):514-528.doi:10.1016/j.pcad.2015.11.003.
[21]Lim CS,Nam HJ,Lee J,et al.PKCα-mediated phosphorylation of LSD1 is required for presynaptic plasticity and hippocampal learning and memory[J].Sci Rep,2017,7(1):4912.doi:10.1038/s41598-017-05239-7.2018-09-262019-02-27
[2]De la Pe?a JB,Lee HL,Yoon SY,et al.The involvement of magnoflorine in the sedative and anxiolytic effects of Sinomeni Caulis et Rhizoma in mice[J].J Nat Med,2013,67(4):814-821.doi:10.1007/s11418-013-0754-3.
[3]Chatterjee P,Roy D,Rathi N.Epigenetic drug repositioning for Alzheimer′s disease based on epigenetic targets in human interactome[J].J Alzheimers Dis,2018,61(1):53-65.doi:10.3233/JAD-161104.
[4]衣泰龙,涂悦,张赛,等.基于双向电泳的应激性抑郁症模型大鼠蛋白质组学研究[J].天津医药,2017,45(8):825-829.Yi TL,Tu Y,Zhang S,et al.Proteomics study of stress-induced depression in rat model based on the two dimensional electrophoresis[J].Tianjin Med J,2017,45(8):825-829.doi:10.11958/20170656.
[5]Cheng K,Li J,Yang D,et al.2D-gel based proteomics unravels neurogenesis and energetic metabolism dysfunction of the olfactory bulb in CUMS rat model[J].Behav Brain Res,2016,313:302-309.doi:10.1016/j.bbr.2016.05.026.
[6]Qiao H,Li MX,Xu C,et al.Dendritic spines in depression:what we learned from animal models[J].Neural Plast,2016:8056370.doi:10.1155/2016/8056370.
[7]Song P,Zhang Y,Ma G,et al.Gastrointestinal absorption and metabolic dynamics of jujuboside A,a saponin derived from the seed of Ziziphus jujube[J].J Agric Food Chem,2017,65(38):8331-8339.doi:10.1021/acs.jafc.7b02748.
[8]Chimire S,Kim MS.Jujube(Ziziphus jujuba Mill.)fruit feeding extends lifespan and increases tolerance to environmental stresses by regulating aging-associated gene expression in Drosophila[J].Biogerontology,2017,18(2):263-273.doi:10.1007/s10522-017-9686-8.
[9]杨奕.酸枣仁生物碱部分质量控制方法及其抗抑郁作用机制初步研究[D].天津:天津医科大学,2013.Yang Y.The study of the quality control and primary research of depression-relieving and spirit-quieting mechanism of total alkloids in semen Zizyphi Spinosae[D].Tianjin:Tianjin Medical University,2013.
[10]Mo?d?eńE,W?sik A,Romańska I,et al.Antidepressant-like effect of 1,2,3,4-tetrahydroisoquinoline and its methyl derivative in animal models of depression[J].Pharmacol Rep,2017,69(3):566-574.doi:10.1016/j.pharep.2017.01.032.
[11]Haque MA,Jantan I,Harikrishnan H,et al.Magnoflorine enhances LPS-activated pro-inflammatory responses via MyD88-dependent pathways in U937 macrophages[J].Planta Med,2018,84(17):1255-1264.doi:10.1055/a-0637-9936.
[12]Tan HL,Chan KG,Pusparajah P,et al.Rhizoma Coptidis:Apotential cardiovascular protective agent[J].Front Pharmacol,2016,7:362.doi:10.3389/fphar.2016.00362.
[13]宋伟.酸枣仁生物碱与黄酮配伍发挥解郁安神作用机制的研究[D].天津:天津医科大学,2018.Song W.Study on mechanism of depression-relieving and spirit-quieting based on compatibility of the alkaloids and flavonoids from SZS[D].Tianjin:Tianjin Medical University,2018.
[14]马全明,王荣,王亚峰.大鼠血浆中木兰花碱的HPLC法测定及药动学研究[J].中国药房,2011,22(35):3280-3282.Ma QM,Wang R,Wang YF.Determination of magnoflorine in rat plasma by HPLC and pharmacokinetic study[J].China Pharmacy,2011,22(35):3280-3282.
[15]Puglia C,Frasca G,Musumeci T,et al.Curcumin loaded NLCinduces histone hypoacetylation in the CNS after intraperitoneal administration in mice[J].Eur J Pharm Biopharm,2012,81(2):288-293.doi:10.1016/j.ejpb.2012.03.015.
[16]Cao C,Wu H,Vasilatos SN,et al.HDAC5-LSD1 axis regulates antineoplastic effect of natural HDAC inhibitor sulforaphane in human breast cancer cells[J].Int J Cancer,2018,143(6):1388-1401.doi:10.1002/ijc.31419.
[17]Wang P,Fan F,Li X,et al.Riboflavin attenuates myocardial injury via LSD1-mediated crosstalk between phospholipid metabolism and histone methylation in mice with experimental myocardial infarction[J].J Mol Cell Cardiol,2018,115:115-129.doi:10.1016/j.yjmcc.2018.01.006.
[18]Kim D,Nam HJ,Lee W,et al.PKCα-LSD1-NF-κB-signaling cascade is crucial for epigenetic control of the inflammatory response[J].Mol Cell,2018,69(3):398-411.doi:10.1016/j.molcel.2018.01.002.
[19]Schulz-Fincke J,Hau M,Barth J,et al.Structure-activity studies on N-Substituted tranylcypromine derivatives lead to selective inhibitors of lysine specific demethylase 1(LSD1)and potent inducers of leukemic cell differentiation[J].Eur J Med Chem,2018,144:52-67.doi:10.1016/j.molcel.2018.01.002.
[20]Teply RM,Packard KA,White ND,et al.Treatment of depression in patients with concomitant cardiac disease[J].Prog Cardiovasc Dis,2016,58(5):514-528.doi:10.1016/j.pcad.2015.11.003.
[21]Lim CS,Nam HJ,Lee J,et al.PKCα-mediated phosphorylation of LSD1 is required for presynaptic plasticity and hippocampal learning and memory[J].Sci Rep,2017,7(1):4912.doi:10.1038/s41598-017-05239-7.2018-09-262019-02-27