利血平预处理抑制低频电刺激对大鼠脑组织Hsp-70和c-fos水平的上调
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摘要
目的观察利血平对低频电刺激引起的应激的影响。方法给予Wistar大鼠利血平预处理再给予低频电刺激,即刻剥取脑组织,用酶联免疫吸附法(ELISA)和免疫组织化学(IHC)检测脑组织中c-fos和Hsp-70基因的表达。结果电刺激组大脑前额叶皮层(PFC)、中脑腹侧背盖区(VTA)、海马(Hipp)组织中Hsp-70和c-fos水平比对照组相应区域均显著升高,免疫反应阳性细胞数明显增多;利血平处理使电刺激产生的Hsp-70和c-fos水平的增加和免疫反应阳性细胞数的增加均明显减少。结论利血平预处理可减轻低频电刺激引起的应激损伤。
Objective To determine the effect of reserpine on stress induced by low-frequency electrical stimulation. Methods Wistar rats, with or without reserpine pretreatment, were stimulated with low-frequency current. Their brain were removed immediately after the stimulation to measure Hsp-70 and c-fos expression using enzyme-linked immunosorbent assay(ELISA) and immunohistochemistry. Results Compared to control group, rats after electrical stimulation showed increased Hsp-70 levels in prefrontal cortex(PFC), midbrain ventral tegmental area(VTA), and hippocampus(Hipp) by 1116.67%, 1105.24% and 1013.76% respectively. c-fos levels also increased by 434.94%, 419.06% and 363.49% in corresponding areas. The number of Hsp-70 and c-fos positive cells was significantly higher in stimulated rats than in control group. Reserpine pre-treatment significantly reduced the elevated Hsp-70 and c-fos expression levels as well as the number of IHC stain positive cells. Conclusion Reserpine pretreatment may likely reduce the stress damage induced by low-frequency electrical stimulation.
Objective To determine the effect of reserpine on stress induced by low-frequency electrical stimulation. Methods Wistar rats, with or without reserpine pretreatment, were stimulated with low-frequency current. Their brain were removed immediately after the stimulation to measure Hsp-70 and c-fos expression using enzyme-linked immunosorbent assay(ELISA) and immunohistochemistry. Results Compared to control group, rats after electrical stimulation showed increased Hsp-70 levels in prefrontal cortex(PFC), midbrain ventral tegmental area(VTA), and hippocampus(Hipp) by 1116.67%, 1105.24% and 1013.76% respectively. c-fos levels also increased by 434.94%, 419.06% and 363.49% in corresponding areas. The number of Hsp-70 and c-fos positive cells was significantly higher in stimulated rats than in control group. Reserpine pre-treatment significantly reduced the elevated Hsp-70 and c-fos expression levels as well as the number of IHC stain positive cells. Conclusion Reserpine pretreatment may likely reduce the stress damage induced by low-frequency electrical stimulation.
引文
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[15]Yang L,Huang Y,Wang FL,et al.Effects of cold stress on immune system and Hsp70 m RNA expression in Altay sheep.Chin J Zool,2015,50(2):300-305.
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[19]Yu FR,Lian XZ,Xie MR,et al.Effects of immediate stimulation on single nucleotide and catecholamine level in Chinese tree shrews.Adv Materi Res,2014,998(7):233-237
[20]俞发荣,谢明仁,连秀珍,等.低频电刺激对不同动物血内激素水平的影响.中国实验动物学报,2015,21(3):82-84.
[2]Morgan S,Greenberg ME,The regulation and function of c-fosnd other l EGS in the neurous system.Neuron,1990,4(2):477-485.
[3]尹柏双,石星星,李小波.阿替美唑对小型猪特异性麻醉剂诱导大鼠大脑皮质内c-fos蛋白表达的影响.畜牧兽医学报,2012,43(5):821-825.
[4]杜艳,赵利华,吴海标,等.艾炷灸对D-半乳糖衰老小鼠学习记忆能力和大脑皮质、海马区c-fos蛋白表达的影响.时珍国医国药,2011,22(1):46-48.
[5]Penny ML,Bruno SB,Cornelius J,et al.The effects of osmotic stimulation and water availability on c-Fos and Fos B staining in the supraoptic and paraventricular nuclei of the hypothalamus.Exp Neurol,2005,194(1):191-202.
[6]Meng X,Li YT,Hu W,et al.Effect of sevoflurane labor analgesia on the expression of c-fos in brain tissue of neonatal rats with hypoxic-ischemic brain damage.J Xinxiang Med Univ,2017,34(1):26-28.
[7]Zhu MH,Lu WH,Li JL,et al.Effects of naloxone injected into cisterna magna on expression of c-Fos proteins and c-Fos m RNA in brain tissues of rats following cardiopulmonary resuscition.J Guangzhou Med,2016,47(2):4-7.
[8]Zhang XQ,Wu MY,Liu CY,et al.Effect of transcranial magnetic stimulation on the expression of c-Fos and brain-derived neurotrophic factor of the cerebral cortex in rats with cerebral infarct.J Huazhong Univ Sci Technol Med Sci,2007,27(4):415-418.
[9]李伟,于建设,辛学东,等.不同血型大鼠电刺激后c-fos基因表达的差异.临床麻醉学杂志,2012,28(8):807-808.
[10]Du YJ,Song J,ZHou H,et al.A study on the mechanism for neuron protection by heat shock protein after pre-acupuncture.J World Acup–Moxib,2014,24(1):32-38.
[11]李琦,潘芳,江虹,等.不同应激方式与时程对大鼠海马CA3区HSP70表达及血清皮质醇含量的影响.山东大学学报(医学版),2005,43(11):1001-1005,1009.
[12]Li Y,Li Y,Dong WW.The effect of focal cerebral ischemia/reperfusion and electro-acupuncture on expression of Hsp70 in brain tissue of Wistar rats.Brain Pathol Brain Pathol,2000,10(4):565-565.
[13]Deng ZK,Dong WW.Effect of cerebellar fastigial nucleus electrical stimulation on expression of heat shock protein70 in somstosensory cortex and basal ganglia of rats.J Chin Neurosci,2002,18(2):499-513.
[14]Xavier DR,Tomàs S,Pierre YF,et al.In vivo imaging of induction of heat-shock protein-70 gene expression with fluorescence reflectance imaging and intravital confocal microscopy following brain ischaemia in reporter mice.Eur J Nucl Med Mol Imag,2013,40(3):426-438.
[15]Yang L,Huang Y,Wang FL,et al.Effects of cold stress on immune system and Hsp70 m RNA expression in Altay sheep.Chin J Zool,2015,50(2):300-305.
[16]Yang TT,Gan L,Zhou Y.Effect of thermal stimulation on expression of Hsp70 m RNA in osteoblasts&its medium’s effect on proliferation of osteoclasts in mice.Prog Mod Biomed,2010,10(19):3639-3642.
[17]Irina VE,Larisa EN,Irina VR,et al.Exogenous protein Hsp70/Hsc70 can penetrate into brain structures and attenuate the severity of chemically-induced seizures.J.Neurochem,2010,115(4):1035–1044.
[18]Valbonesi P,Ricci L,Franzellitti S,et al.Effects of cadmium on MAPK signalling pathways and HSP70 expression in a human trophoblast cell line.Placenta,2008,29(8):725-733.
[19]Yu FR,Lian XZ,Xie MR,et al.Effects of immediate stimulation on single nucleotide and catecholamine level in Chinese tree shrews.Adv Materi Res,2014,998(7):233-237
[20]俞发荣,谢明仁,连秀珍,等.低频电刺激对不同动物血内激素水平的影响.中国实验动物学报,2015,21(3):82-84.