脑髓康保护糖氧剥夺诱导损伤脑微血管内皮细胞的机制研究
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摘要
目的:探讨脑髓康对糖氧剥夺(OGD)诱导损伤大鼠脑微血管内皮细胞(RBMEC)保护作用的机制原理。方法:构建RBMEC损伤模型,按实验分组给予含相应血清的培养液处理,应用CCK8法检测细胞存活率,流式细胞术检测细胞凋亡率,分光光度法检测细胞中氧化应激相关因子超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量,Western Blot法检测细胞中内质网应激相关因子CCAAT/增强子结合蛋白同源蛋白(CHOP)和半胱胺酸蛋白酶蛋白-12(Caspase 12)的表达情况。结果:与糖氧剥夺组比较,脑髓康能够显著提高OGD诱导损伤RBMEC的存活率(P<0.01),并且显著抑制其凋亡率(P<0.01);能够缓解OGD诱导损伤RBMEC中的氧化应激,显著提高细胞中SOD和GSH-Px的活性(P<0.01,P<0.05),同时降低细胞中MDA的含量(P<0.01);此外,脑髓康显著降低了内质网应激相关因子CHOP和Caspase 12的表达水平(P<0.01)。结论:脑髓康可能通过缓解氧化应激以及抑制内质网应激介导的细胞调亡途径,对OGD诱导损伤RBMEC发挥保护作用。
Objective: To elucidate the mechanisms of Naosuikang in protecting injury of rat brain microvascular endothelial cells(RBMEC) induced by oxygen-glucose deprivation. Methods: RBMEC model was induced by oxygen-glucose deprivation. Corresponding drug serum was administered to different groups. Subsequently, cell survival rate and apoptosis rate were measured by CCK8 and flow cytometry respectively. We also detected the concentration of oxidative stress factors such as superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) and malondialdehyde(MDA), and the protein expression of endoplasmic reticulum stress-related factors CCAAT/enhancer-binding protein homologous protein(CHOP) and Caspase-12 were measured by Western Blot. Results: Naosuikang can significantly elevate the cell survival rate(P<0.01) and diminish the apoptosis rate(P<0.01) in oxygen-glucose deprivation induced injury of RBMEC. In the same time, the levels of SOD(P<0.01) and GSHPx(P<0.05) were elevated and the level of MDA(P<0.01) was reduced by Naosuikang Decoction. What's more, Naosuikang could significantly decrease the levels of CHOP(P<0.01) and Caspase 12(P<0.01). Conclusion: Naosuikang might protect oxygen-glucose deprivation induced injury of RBMEC through regulating oxidative stress and inhibiting endoplasmic reticulum stress mediated apoptosis pathway.
Objective: To elucidate the mechanisms of Naosuikang in protecting injury of rat brain microvascular endothelial cells(RBMEC) induced by oxygen-glucose deprivation. Methods: RBMEC model was induced by oxygen-glucose deprivation. Corresponding drug serum was administered to different groups. Subsequently, cell survival rate and apoptosis rate were measured by CCK8 and flow cytometry respectively. We also detected the concentration of oxidative stress factors such as superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) and malondialdehyde(MDA), and the protein expression of endoplasmic reticulum stress-related factors CCAAT/enhancer-binding protein homologous protein(CHOP) and Caspase-12 were measured by Western Blot. Results: Naosuikang can significantly elevate the cell survival rate(P<0.01) and diminish the apoptosis rate(P<0.01) in oxygen-glucose deprivation induced injury of RBMEC. In the same time, the levels of SOD(P<0.01) and GSHPx(P<0.05) were elevated and the level of MDA(P<0.01) was reduced by Naosuikang Decoction. What's more, Naosuikang could significantly decrease the levels of CHOP(P<0.01) and Caspase 12(P<0.01). Conclusion: Naosuikang might protect oxygen-glucose deprivation induced injury of RBMEC through regulating oxidative stress and inhibiting endoplasmic reticulum stress mediated apoptosis pathway.
引文
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[2]丰梅,付凌玲,张伟华,等.内质网应激调控细胞自噬和凋亡.中国细胞生物学学报,2018,40(3):455-462
[3]薛小燕,郭小华,李敏,等.神经退行性疾病发病机制研究进展.中国老年学杂志,2015,35(11):3149-3152
[4]田年秀,王晓磊,王涛,等.芪卫颗粒含药血清通过Caspase12途径减轻高糖环境下肾足细胞损伤的机制研究.中华中医药杂志,2018,33(5):1858-1862
[5]蔡浩斌,潘华峰,郑浩涛,等.脑髓康通过调控氧化应激及单胺类神经递质改善VCI大鼠情景记忆能力.中药新药与临床药理,2018,29(5):564-567
[6]孙宇,韩璎,戴建平.血管性认知障碍诊断标准的演变与解读.中国卒中杂志,2017,12(1):13-17
[7]Chaaya M,Phung T K,EIAsmar K,et al.Validation of the arabic rowland universal dementia assessment scale(A-RUDAS)in elderly with mild and moderate dementia.Aging Ment Health,2016,20(8):880-887
[8]郑浩涛,王建军,赖雯雯,等.脑小血管病致认知障碍的中医辨析及脑髓康治疗作用探讨.新中医,2018,50(7):220-222
[9]曹甦,虢周科,邝学媚.脑髓康治疗脑动脉硬化症的临床观察.黑龙江医学,2001(4):265
[10]孔繁鑫,虢周科.脑髓康方配合尤瑞克林治疗非溶栓急性期脑梗死临床观察.山西中医,2016,32(6):6-8+17
[11]秦秀德,李传朋,龚胜兰,等.基于虚和瘀探讨脑髓康对中风的治疗作用.辽宁中医杂志,2017,44(2):260-261
[12]胡勇,巨少华,张荫杰,等.通络醒脑泡腾片对大鼠慢性脑缺血致学习记忆功能损伤的影响.中国中药杂志,2014,39(10):1908-1912
[13]张玥,罗俊,黄能慧,等.灵芝三萜类化合物对AD衰老模型大鼠学习记忆能力和脑能量代谢的影响.食品与生物技术学报,2012,31(7):741-744
[14]徐世军,马涛,张文生,等.解毒益智胶囊对慢性脑缺血性学习记忆障碍大鼠学习记忆和海马突触素表达的影响.中药药理与临床,2011,27(3):93-96
[15]Hetz C,Mollereau B.Disturbance of endoplasmic reticulum proteostasis in neurodegenerative diseases.Nature Reviews Neuroscience,2014,15(4):233-249
[16]Hong J,Kim K,Kim J H,et al.The Role of Endoplasmic Reticulum Stress in Cardiovascular Disease and Exercise.International Journal of Vascular Medicine,2017,2017(5):1-9
[17]Wang M,Kaufman R J.Protein misfolding in the endoplasmic reticulum as a conduit to human disease.Nature,2016,529(7586):326
[18]Oyadomari S,Mori M.Roles of CHOP/GADD153 in endoplasmic reticulum stress.Cell Death&Differentiation, 2004,11(4):381
[19]Bravo R,Gutierrez T,Paredes F,et al.Endoplasmic reticulum:ER stress regulates mitochondrial bioenergetics.International Journal of Biochemistry&Cell Biology,2012,44(1):16-20
[20]Omura T,Kaneko M,Okuma Y,et al.Endoplasmic reticulum stress and Parkinson’s disease:the role of HRD1 in averting apoptosis in neurodegenerative disease.Oxidative Medicine and Cellular Longevity,2013,2013:239854
[21]Gaballah H H,Zakaria S S,Elbatsh M M,et al.Modulatory effects of resveratrol on endoplasmic reticulum stress-associated apoptosis and oxido-inflammatory markers in a rat model of rotenone-induced Parkinson’s disease.Chemico-Biological Interactions,2016,251:10-16