逍遥散对LPS所致大鼠神经损伤的保护作用机制
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摘要
目的:探讨逍遥散对脂多糖(LPS)所致大鼠神经损伤的保护作用,探讨其机制。方法:56只SD大鼠随机分为空白组,假手术组,模型组,阿米替林组(10 mg·kg-1),氟西汀组(10 mg·kg-1),逍遥散高、低剂量组(30,15 g·kg-1),采用侧脑室注射LPS诱导建立神经损伤大鼠模型,连续预防灌胃给药14 d,酶联免疫吸附测定(ELISA)检测大鼠血清脑源性神经营养因子(BDNF)和β-神经生长因子(β-NGF)水平,实时荧光定量聚合酶链式反应(Real-time PCR)与蛋白免疫印迹法(Western blot)检测大鼠海马和皮层部位BDNF,神经生长因子(NGF),原肌球蛋白受体激酶B(Trk B),原肌球蛋白受体激酶A(Trk A),c AMP反应元件结合蛋白(CREB),突触后密度蛋白95(PSD95),突触小泡蛋白(SYP) mRNA或蛋白表达水平。结果:与空白组比较,模型组大鼠血清BDNF,β-NGF含量显著下降(P <0. 01),皮层、海马BDNF,NGF,Trk B,Trk A,CREB mRNA明显下调(P <0. 05,P <0. 01),BDNF,Trk B,CREB,磷酸化c AMP反应元件结合蛋白(p-CREB),PSD95,SYP蛋白表达水平明显下调(P<0. 05,P <0. 01);与模型组比较,逍遥散高、低剂量组大鼠血清BDNF,β-NGF含量明显升高(P <0. 05,P <0. 01),皮层、海马BDNF,NGF,Trk B,Trk A,CREB mRNA表达水平明显上调(P <0. 05,P <0. 01),BDNF,Trk B,CREB,p-CREB,PSD95,SYP蛋白表达水平明显上调(P <0. 05,P <0. 01)。结论:逍遥散对侧脑室注射LPS诱导的大鼠神经损伤有一定保护作用,作用发挥与活化BDNF/NGF-TrkB/Trk A-CREB通路及上调突触蛋白表达有关。
Objective: To investigate the effect and mechanism of Xiaoyaosan on lipopolysaccharide(LPS)-induced nerve injury. Method: The 56 rats were randomly divided into control group,sham group,model group,amitriptyline group(10 mg·kg-1),fluoxetine group(10 mg·kg-1),Xiaoyaosan group high and lowdose(30,15 g·kg-1). The nerve injury model rat were established by LPS injection into lateral ventride,rats were administrated for 14 days by gavage. The levels of brain-derived neurotrophic factor(BDNF) and β-nerve growth factor(β-NGF) in serum were detected by enzyme linked immunosorbent assay(ELISA),and the expressions of BDNF,nerve growth factor(NGF),tropomyosin receptor kinase B(TrkB),tropomyosin receptor kinase A(TrkA),cAMP response element-binding protein(CREB) mRNA in hippocampus and cortex were detected by Real-time PCR. Protoin expression of BDNF,Trk B,CREB,p-CREB,postsynaptic density protein 95(PSD95),synaptophysin(SYP) in hippocampus and cortex were detected by Western blot. Result: Compared with control group,LPS decreased the level of BDNF and β-NGF in serum(P < 0. 01),reduced the expression of BDNF,NGF,Trk B, TrkA,CREB mRNA and the expression of BDNF,Trk B,CREB,Phospho-CREB(pCREB),PSD95,SYP in hippocampus and cortex(P < 0. 05,P < 0. 01). Compared with model group,levels of BDNF and β-NGF in serum in Xiaoyaosan high and low-dose group were increased significantly(P < 0. 05,P <0. 01),and expressions of BDNF,NGF,TrkB,TrkA,CREB,PSD95,SYP in hippocampus and cortex were increased significantly(P < 0. 05,P < 0. 01). Conclusion: Xiaoyaosan has a certain antagonistic effect on LPS inducednerve injury,which suggests that the effect is related to activate BDNF/NGF-TrkB/TrkA-CREB pathway and upregulated the expression of synaptic protein.
Objective: To investigate the effect and mechanism of Xiaoyaosan on lipopolysaccharide(LPS)-induced nerve injury. Method: The 56 rats were randomly divided into control group,sham group,model group,amitriptyline group(10 mg·kg-1),fluoxetine group(10 mg·kg-1),Xiaoyaosan group high and lowdose(30,15 g·kg-1). The nerve injury model rat were established by LPS injection into lateral ventride,rats were administrated for 14 days by gavage. The levels of brain-derived neurotrophic factor(BDNF) and β-nerve growth factor(β-NGF) in serum were detected by enzyme linked immunosorbent assay(ELISA),and the expressions of BDNF,nerve growth factor(NGF),tropomyosin receptor kinase B(TrkB),tropomyosin receptor kinase A(TrkA),cAMP response element-binding protein(CREB) mRNA in hippocampus and cortex were detected by Real-time PCR. Protoin expression of BDNF,Trk B,CREB,p-CREB,postsynaptic density protein 95(PSD95),synaptophysin(SYP) in hippocampus and cortex were detected by Western blot. Result: Compared with control group,LPS decreased the level of BDNF and β-NGF in serum(P < 0. 01),reduced the expression of BDNF,NGF,Trk B, TrkA,CREB mRNA and the expression of BDNF,Trk B,CREB,Phospho-CREB(pCREB),PSD95,SYP in hippocampus and cortex(P < 0. 05,P < 0. 01). Compared with model group,levels of BDNF and β-NGF in serum in Xiaoyaosan high and low-dose group were increased significantly(P < 0. 05,P <0. 01),and expressions of BDNF,NGF,TrkB,TrkA,CREB,PSD95,SYP in hippocampus and cortex were increased significantly(P < 0. 05,P < 0. 01). Conclusion: Xiaoyaosan has a certain antagonistic effect on LPS inducednerve injury,which suggests that the effect is related to activate BDNF/NGF-TrkB/TrkA-CREB pathway and upregulated the expression of synaptic protein.
引文
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[2] Dantzer R,O'Connor J C,Freund G G,et al. From inflammation to sickness and depression:when the immune system subjugates the brain[J]. Nat Rev Neurosci,2008,9(1):46-56.
[3] TANG M M,LIN W J,PAN Y Q,et al. Hippocampal neurogenesis dysfunction linked to depressive-like behaviors in a neuro inflammation induced model of depression[J]. Physiol Behav,2016,161:166-173.
[4] Marlatt M W,Lucassen P J,Van P H. Comparison of neurogenic effects of fluoxetine,duloxetine and running in mice[J]. Brain Res,2010,1341(6):93-99.
[5] Encinas J M,Vaahtokari A,Enikolopov G. Fluoxetine targets early progenitor cells in the adult brain[J]. P Natl Acad Sci USA,2006,103(21):8233-8238.
[6] Hitoshi S,Maruta N,Higashi M,et al. Antidepressant drugs reverse the loss of adult neural stem cells following chronic stress[J]. J Neurosci Res,2007,85(16):3574-3585.
[7]杨靖.逍遥散及其功能药队抗抑郁效应和分子机制研究[D].成都:成都中医药大学,2014.
[8]段富津.方剂学[M].上海:上海科学技术出版社,1995.
[9] Chiou S H,CHEN S J,PENG C H,et al. Fluoxetine up-regulation expression of cellular FLICE-inhibitory protein and inhibits LPS-induced apoptosis in hippocampus-derived stem cell[J]. Biochem Bioph Res Co,2006,343(2):391-400.
[10]王学,叶晓琳,刘小波,等.逍遥散抗抑郁作用的研究现状[J].中国实验方剂学杂志,2018,24(16):212-220.
[11] Cohen-Cory S. The developing synapse:construction and modulation of synapticstructures and circuits[J].Science,2002,298(5594):770-776.
[12] Bathina S,Das U N. Brain-derived neurotrophic factor and its clinical implications[J]. Arch Med Sci,2015,11(6):1164-1178.
[13] Devry J,Vanmierlo T,Martinez-Martinez P,et al.Trk B in the hippocampus and nucleus accumbens differentially modulates depression-like behavior in mice[J]. Behav Brain Res,2016,296:15-25.
[14] LU SH,YANG Y,LIU S J. An investigation on the division of neuronal PC12 cells induced by nerve growth factor[J]. Acta Physiol Sinica, 2005, 57(5):552-556.
[15] Riccio A,Alvania R S,Lonze B E,et al. A nitric oxide signaling pathway controls creb-mediated gene expression in neurons[J]. Mol Cell,2009,21(2):283-294.
[16] PAN W, HAN S, LIN K, et al. Effects of dihydrotestosterone on synaptic plasticity of the hippocampus in mild cognitive impairment male SAMP8mice[J]. Exp Ther Med,2016,12(3):1455-1463.
[17] XU J Q,Winter F D,Farrokhi C,et al. Neuregulin 1improves cognitive deficits and neuropathology in an Alzheimer's disease model[J]. Sci Rep,2016,doi:10.1038/srep31692.
[18] Schmitt U,Tanimoto N,Seeliger M,et al. Detection of behavioral alterations and learning deficits in mice lacking synaptophysin[J]. Neuroscience,2009,162(2):234-243.
[19] WU X,YANG G L,DU Y F,et al. Neuroprotective effects of int-777 against Aβ1-42-induced cognitive impairment,neuroinflammation,apoptosis,and synaptic dysfunction in mice[J]. Brain Behav Immun,2018,doi:10. 1016/j. bbi. 2018. 06. 018.