逍遥散对慢性应激损伤模型大鼠海马区PLA2G5的影响
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摘要
目的:研究逍遥散对慢性应激损伤模型大鼠海马区V型分泌型磷脂酶(PLA2G5)基因表达及翻译的影响,探讨其改善慢性应激损伤的可能机制。方法:将120只Wistar雄性大鼠随机分为空白对照组、慢性应激模型组、逍遥散干预组、开克对照组4组,每组30只。造模方法采用慢性温和不可预知应激(CUMS),逍遥散干预组和开克对照组在造模同时分别给予逍遥散10mL/kg、开克10 mL/kg。造模时间分别为21、28、35天。采用荧光定量RT-PCR及Western-Blot检测方法测定各组大鼠海马区PLA2G5基因及其蛋白表达情况。结果:慢性应激模型组大鼠海马区PLA2G5基因mRNA表达及其相应蛋白显著下调。逍遥散可从基因水平同时纠正这种下调的表达变化。结论:PLA2G5的表达变化参与了慢性应激损伤过程,逍遥散对慢性应激所致的损伤可能通过调节这一基因发挥保护性作用。
Objective:To research the effect of Xiaoyao powder on expression and translation of PLA2 G5 gene in hippocampal area of model rats with chronic stress injury,and explore the possible mechanism of improving chronic stress injury. Methods:120 male Wistar rats were randomly divided into blank control group,chronic stress model group,treatment group of Xiaoyao powder and control group of fluoxetine hydrochloride,30 rats in each group. Chronic mild and unpredictable stress(CUMS) was adopted as modeling method. Treatment group of Xiaoyao powder and control group of fluoxetine hydrochloride were given respectively 10 mL/Kg of Xiaoyao powder and 10 mL/Kg of fluoxetine hydrochloride at the same time of molding. Molding time was respectively 21,28 and35 days.The expressions of PLA2 G5 gene and its protein in hippocampus of rats in each group were detected by fluorescence quantitative RT-PCR and Western-Blot methods.Results:m RNA expressions of PLA2 G5 gene and its corresponding protein in hippocampal area of rats in chronic stress model group were regulated down significantly.Xiaoyao powder could correct the change of this down-regulated expression simultaneously from genetic level.Conclusion:The change of A2 PLA2 G5 expression participated in the process of chronic stress injury. Xiaoyao powder may play a protective role to chronic stress injury by regulating this gene.
Objective:To research the effect of Xiaoyao powder on expression and translation of PLA2 G5 gene in hippocampal area of model rats with chronic stress injury,and explore the possible mechanism of improving chronic stress injury. Methods:120 male Wistar rats were randomly divided into blank control group,chronic stress model group,treatment group of Xiaoyao powder and control group of fluoxetine hydrochloride,30 rats in each group. Chronic mild and unpredictable stress(CUMS) was adopted as modeling method. Treatment group of Xiaoyao powder and control group of fluoxetine hydrochloride were given respectively 10 mL/Kg of Xiaoyao powder and 10 mL/Kg of fluoxetine hydrochloride at the same time of molding. Molding time was respectively 21,28 and35 days.The expressions of PLA2 G5 gene and its protein in hippocampus of rats in each group were detected by fluorescence quantitative RT-PCR and Western-Blot methods.Results:m RNA expressions of PLA2 G5 gene and its corresponding protein in hippocampal area of rats in chronic stress model group were regulated down significantly.Xiaoyao powder could correct the change of this down-regulated expression simultaneously from genetic level.Conclusion:The change of A2 PLA2 G5 expression participated in the process of chronic stress injury. Xiaoyao powder may play a protective role to chronic stress injury by regulating this gene.
引文
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[23]李肖,宫文霞,周玉枝,等.逍遥散中抗抑郁有效成分及其作用机制研究进展[J].中草药,2015,46(20):3 109-3 116.
[2]马素亚,郑俊超,程丹,等.锁阳乙酸乙酯提取物改善慢性应激认知功能障碍MAPK/ERK1/2通路机制[J].世界中西医结合杂志,2017,12(10):1 381-1 385.
[3]金钟晔.逍遥散对慢性束缚应激肝郁脾虚证焦虑模型大鼠下丘脑P38MAPK信号通路的影响[D].北京中医药大学,2016:19-21.
[4]邱娟.柴胡疏肝散及拆方对抑郁模型大鼠行为学和脑组织p38MAPK、ERK5表达的影响[D].长沙:中南大学,2014:46-49.
[5]Yamaguchi M,Zacharia J,Laidlaw TM,et al.PLA2G5regulates transglutaminase activity of human IL-4-activated M2 macrophages through PGE2 generation[J].J Leukoc Biol,2016,100(1):131-141.
[6]Silva-Filho JL,Peruchetti DB,Moraes-Santos F,et al.Group V Secretory Phospholipase A2 Is Involved in Tubular Integrity and Sodium Handling in the Kidney[J].PLoS One,2016,11(1):e0147785.
[7]Menschikowski M,Hagelgans A,Nacke B,et al.Epigenetic control of group V phospholipase A2 expression in human malignant cells[J].Tumour Biol,2016,37(6):8 097-8 105.
[8]Murakami M,Kudo I.Phospholipase A2[J].J Biochem,2002,131(3):285-292.
[9]P r u z a n s k i W,L a m b e a u L,L a z d u n s k y M,e t a l.Differential hydrolysis of molecular species of lipoprotein phosphatidylcholine by groups IIA,V and X secretory phospholipases A2[J].Biochim Biophys Acta,2005,1736(1):38-50.
[10]任洁.分泌型磷脂酶A2基因多态性与冠心病遗传易感性的研究[D].蚌埠:蚌埠医学院,2015:24-26.
[11]Creighton J.Factors controlling vascular permeability:Transmitting mechanical signals.Focus on"Mechanical induction of GroupⅤphospholipase A2 causes lung inflammation and acute lung injury"[J].Am J physiol Lung cell Mol physiol,2013,305(4):279-281.
[12]Meliton A Y,Munoz N M.Meliton L N,et al.Mechanical induction of group V phospholipase A2 causes lung inflammation and acute lung injury[J].Am J Physiol Lung Cel Mol Physiol,2013,304(10):689-700.
[13]Degousee N,Kelvin D J,Geisslinger G,et al.Croup VPhospholipase A2 in Bone Marrow-derived Myeloid Cells and Bronchial Epithelial Cells Promotes Bacterial Clearance after Escherichia coli Pneumonia[J].Journal of Biological Chemistry,2011,286(41):35 650-35 662.
[14]Yamaguchi M,Samuchiwal SK,Quehenberger O,et al.Macrophages regulate lung ILC2 activation via Pla2g5-dependent mechanisms[J].Mucosal Immunol,2018,11(3):615-626.
[15]Bostrom MA,Boyanovsky BB,Jordan CT,et al.Croup Vsecretory phospholipaseA2 promotes atherosclerosis:evidence from genetically altered mice[J].Arterioscler Thromb Vasc Biol,2007,27(3):600-606.
[16]Masuda S,Murakami M,Mitsuishi M,et al.Expression of secretory phospholipase A2 enzymes in lungs of humans with pneumonia and their potential prostaglandin-synthetic function in human lung-derived cells[J].Biochem J,2005,387(Pt1):27-38.
[17]富文俊.逍遥散对慢性应激损伤大鼠HPA轴负反馈功能及GR、NR亚型调节机制的研究[D].广州:广州中医药大学,2009:37.
[18]孙琪.逍遥散对慢性应激损伤大鼠海马神经细胞的影响及作用机制研究[D].广州:广州中医药大学,2010:89.
[19]Brunzell D H,Russell D S,Picciotto M R.In vivo nicotine treatment regulates mesocorticolimbic CREB and ERK signaling in C57Bl/6J mice[J].J Neurochem,2003,84(6):1 431-1 441.
[20]Thomas G M,Huganir R L.MAPK cascade signalling and synaptic plasticity[J].Nat Rev Neurosci,2004,5(3):173-183.
[21]潘苗,张三强,裴丽粉,等.慢性应激抑郁大鼠海马CA1神经元突触可塑性研究[J].中国神经精神疾病杂志,2012(11):672-676.
[22]Lundin L,Ronnstrand L,Cross M,et al.Differential tyrosine phosphorylation of fibroblast growth factor(FGF)receptor-1and receptor proximal signal transduction in response to FGF-2 and heparin[J].Experimental cell research,2003,287(1):190-198.
[23]李肖,宫文霞,周玉枝,等.逍遥散中抗抑郁有效成分及其作用机制研究进展[J].中草药,2015,46(20):3 109-3 116.