[1]季禾,栗延伟,谭军,等.腺苷预处理对脑缺血再灌注损伤大鼠脑组织中CCAAT/增强子结合蛋白同源蛋白表达的影响[J].新乡医学院学报,2023,40(2):101-106.[doi:10.7683/xxyxyxb.2023.02.001]
　JI He,LI Yanwei,TAN Jun,et al.Effect of adenosine pretreatment on expression of CCAAT/enhancer binding protein homologous protein in brain tissue of rats with cerebral ischemia-reperfusion injury[J].Journal of Xinxiang Medical University,2023,40(2):101-106.[doi:10.7683/xxyxyxb.2023.02.001]
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腺苷预处理对脑缺血再灌注损伤大鼠脑组织中CCAAT/增强子结合蛋白同源蛋白表达的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
40卷
期数:
2023年2期
页码:
101-106
栏目:
基础研究
出版日期:
2023-02-05

文章信息/Info

Title:
Effect of adenosine pretreatment on expression of CCAAT/enhancer binding protein homologous protein in brain tissue of rats with cerebral ischemia-reperfusion injury
作者:
季禾1; 栗延伟2; 谭军2; 曹武璟1; 张振祥3
（1.新乡医学院第三临床学院，河南　新乡　453003;2.新乡医学院第三附属医院神经内科，河南　新乡　453003;3.新乡医学院第三附属医院呼吸内科，河南　新乡　453003）
Author(s):
JI He1; LI Yanwei2; TAN Jun2; CAO Wujing1; ZHAGN Zhenxiang3
（1.The Third Clinical College of Xinxiang Medical University,Xinxiang 453003,Henan Province,China;2.Department of Neurology,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China;3.Department of Respiratory,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China）
关键词:
脑缺血再灌注损伤; 腺苷; CCAAT/增强子结合蛋白同源蛋白; 内质网应激; 细胞凋亡
Keywords:
cerebral ischemia-reperfusion injury; adenosine; CCAAT/enhancer binding protein homologous protein; endoplasmic reticulum stress; cell apoptosis
分类号:
R743.3
DOI:
10.7683/xxyxyxb.2023.02.001
文献标志码:
A
摘要:
目的　观察腺苷预处理（AP）对脑缺血再灌注损伤大鼠脑组织中CCAAT/增强子结合蛋白同源蛋白（CHOP）表达的影响，探讨AP对脑缺血再灌注损伤大鼠的神经保护作用机制。
方法　将36只健康雄性Sprague Dawley大鼠按随机数字表法分为假手术组、缺血再灌注（IR）组和AP组，每组12只。AP组大鼠术前3 d腹腔注射1.5 mg·kg-1腺苷注射液，每日1次；假手术组和IR组大鼠术前3 d腹腔注射等量生理盐水，每日1次。IR组及AP组大鼠采用线栓法制备大脑中动脉栓塞（MCAO）模型，并于栓塞2 h后拔出栓线恢复血供制备IR模型；假手术组大鼠不插入栓线，余处理同IR组及AP组。于再灌注24 h时，根据5分制神经行为学评分标准对3组大鼠进行神经功能缺损评分；然后，断头处死3组大鼠，取脑组织；应用苏木精-伊红染色法观察3组大鼠脑组织病理学改变，实时荧光定量聚合酶链式反应法检测3组大鼠脑组织中CHOP mRNA相对表达量，免疫组织化学法检测3组大鼠脑组织中CHOP蛋白阳性表达情况，脱氧核糖核苷酸末端转移酶介导的缺口末端标记法检测3组大鼠脑组织细胞凋亡情况。
结果　3组大鼠神经功能缺损评分比较差异有统计学意义（F=157.923，P<0.05）；IR 组和AP组大鼠神经功能缺损评分显著高于假手术组(t=17.663、7.133，P<0.05)，AP组大鼠神经功能缺损评分显著低于IR 组(t=-10.530，P<0.05)。假手术组大鼠脑组织神经元形态结构正常，细胞核完整，间质无水肿；IR组大鼠脑组织神经元数目减少、细胞核溶解、出现空泡，间质水肿，组织疏松；AP组大鼠神经元损伤程度显著低于IR组。3组大鼠脑组织中CHOP mRNA相对表达量比较差异有统计学意义（F=2 989.751，P<0.05）；IR组和AP组大鼠脑组织中CHOP mRNA相对表达量显著高于假手术组 (t=75.514、23.502，P<0.05)，AP组大鼠脑组织中CHOP mRNA相对表达量显著低于IR 组(t=-52.171，P<0.05)。3组大鼠脑组织中CHOP 蛋白阳性细胞表达率比较差异有统计学意义（F=2 257.096，P<0.05）；IR组和AP组大鼠脑组织中CHOP 蛋白阳性细胞表达率显著高于假手术组(t=65.927、21.744，P<0.05)，AP组大鼠脑组织中CHOP 蛋白阳性细胞表达率显著低于IR组(t=-44.183，P<0.05)。3组大鼠脑组织细胞凋亡率比较差异有统计学意义（F=1 519.372，P<0.05）；IR组和AP组大鼠脑组织细胞凋亡率显著高于假手术组(t=5.512、2.693，P<0.05)，AP组大鼠脑组织细胞凋亡率显著低于IR组(t=-2.818，P<0.05)。
结论　AP可通过抑制CHOP的表达减轻脑缺血再灌注损伤。
Abstract:
Objective　To observe the effect of adenosine pretreatment (AP) on the expression of CCAAT/enhancer binding protein homologous protein (CHOP) in brain tissue of rats with cerebral ischemia-reperfusion injury,and to explore the neuroprotective mechanism of AP on cerebral ischemia-reperfusion injury in rats.
Methods　Thirty-six healthy male Sprague Dawley rats were divided into sham operation group,ischemia reperfusion(IR) group and AP group according to random number table method,with 12 rats in each group.The rats in the AP group were intraperitoneally injected with 1.5 mg·kg-1 adenosine injection on three days before surgery,once a day;the rats in the sham operation group and IR group were intraperitoneally injected with the same amount of normal saline on three days before surgery,once a day.The middle cerebral artery occlusion(MCAO) models were established by wire embolization in the IR group and AP group,and the thread was pulled out at 2 hours after embolization to prepare the IR models.The rats in the sham operation group were not inserted with wire embolization,and the residual treatments were same as those in the IR group and AP group.At 24 hours of reperfusion,the rats in the three groups were performed with the neurological impairment score according to the 5-point neurobehavioral scale.Then,the rats in the three groups were sacrificed,and the brain tissues of the rats were taken;the histopathological changes in brain tissues of rats in the three groups were observed by hematoxylin-eosin,the relative expression of CHOP mRNA in the brain tissues of rats in the three groups was detected by quantitative real-time polymerase chain reaction,the positive expression of CHOP protein in the brain tissues of rats in the three groups was detected by immunohistochemistry,and the cell apoptosis in the brain tissues of rats in the three groups was detected by TdT mediated dUTP nick end labeling method.
Results　There was significant difference in neurological impairment score of rats among the three groups (F=157.923,P<0.05).The neurological impairment score of rats in the IR group and AP group was significantly higher than that in the sham operation group (t=17.663,7.133;P<0.05),and the neurological impairment score of rats in the AP group was significantly lower than that in the IR group (t=-10.530,P<0.05).In the sham operation group,the morphology and structure of brain neurons of rats were normal,the nuclei were intact,and the interstitial edema was not observed.In the IR group,the number of neurons decreased,the nuclei dissolved,the vacuoles appeared,the interstitial edema was observed and the tissue was loose.The injury degree of neurons of rats in the AP group was significantly lower than that in the IR group.There was significant difference in the relative expression level of CHOP mRNA in brain tissues of rats among the three groups (F=2 989.751,P<0.05).The relative expression level of CHOP mRNA in brain tissues of rats in the IR group and AP group was significantly higher than that in the sham operation group (t=75.514,23.502;P<0.05),and the relative expression level of CHOP mRNA in brain tissues of rats in the AP group was significantly lower than that in the IR group (t=-52.171,P<0.05).There was significant difference in the rate of CHOP protein positive cells in brain tissues of rats among the three groups(F=2 257.096,P<0.05).The rate of CHOP protein positive cells in brain tissue of rats in the IR group and AP group was significantly higher than that in the sham operation group (t=65.927,21.744;P<0.05),and the rate in CHOP protein positive cells in brain tissue of rats in the AP group was significantly lower than that in the IR group (t=-44.183,P<0.05).There was significant difference in the cell apoptosis rate in brain tissue of rats among the three groups (F=1 519.372,P<0.05).The cell apoptosis rate in brain tissue of rats in the IR group and AP group was significantly higher than that in the sham operation group (t=5.512,2.693;P<0.05),and the cell apoptosis rate in brain tissue of rats in the AP group was significantly lower than that in the IR group (t=-2.818,P<0.05).
Conclusion　AP can reduce cerebral ischemia-reperfusion injury by inhibiting the expression of CHOP.

参考文献/References:

［1］　《中国卒中中心报告 2020》编写组.《中国卒中中心报告2020》概要［J］.中国脑血管病杂志,2021,18(11):737-743.
REPORT ON STROKE CENTER IN CHINA WRITING GROUP.Brief report on stroke center in China，2020［J］.Chin J Cerebrovasc Dis,2021,18(11):737-743.
［2］　王春娟,霍晓川,冀瑞俊,等.急性缺血性卒中再灌注治疗医疗质量评价与改进专家建议［J］.中国卒中杂志,2021,16(7):705-715.
WAND C J,HUO X C,JI R J,et al.Consensus on evaluation and improvement of healthcare quality of acute ischemic stroke reperfusion therapy［J］.Chin J Stroke,2021,16(7):705-715.
［3］　庞艳利,许二妮,尉娜,等.腺苷预处理对大鼠脑缺血再灌注损伤后Caspase-3的表达及细胞凋亡的影响［J］.山东医药,2020,60(13):42-45.
PANG Y L,XU E N,WEI N,et al.Effects of adenosine preconditioning on expression of Caspase-3 and apoptosis of rats after cerebral ischemia-reperfusion injury［J］.Shandong Med J,2020,60(13):42-45.
［4］　SUN X,LIU H,SUN Z,et al.Acupuncture protects against cerebral ischemia-reperfusion injury via suppressing endoplasmic reticulum stress-mediated autophagy and apoptosis［J］.Mol Med,2020,26(1):105.
［5］　XU F,MA R,ZHANG G,et al.Estrogen and propofol combination therapy inhibits endoplasmic reticulum stress and remarkably attenuates cerebral ischemia-reperfusion injury and OGD injury in hippocampus［J］.Biomed Pharmacother,2018,108:1596-1606.
［6］　李兆珍,张丹参.脑缺血再灌注损伤相关机制的研究进展［J］.神经药理学报,2020,10(6):60-63.
LI Z Z,ZHANG D S.Research progress on related mechanisms of cerebral ischemia-reperfusion injury［J］.Acta Neuropharmacol,2020,10(6):60-63.
［7］　屈生彪,王勇.内质网应激在脑缺血再灌注损伤中作用［J］.华夏医学,2018,31(1):170-174.
QU S B,WANG Y.Role of endoplasmic reticulum stress in cerebral ischemia-reperfusion injury［J］.Acta Med Sinica,2018,31(1):170-174.
［8］　张翼飞,郑辑英,李东芳,等.内质网应激与脑缺血再灌注损伤研究进展［J］.临床医药实践,2017,26(5):384-387.
ZHANG Y F,ZHENG J Y,LI D F,et al.Research progress of endoplasmic reticulum stress and cerebral ischemia-reperfusion injury［J］.Proceed Clin Med,2017,26(5):384-387.
［9］　王雅男,李红杰,杨一飞,等.腺苷受体激动剂对心肌缺血再灌注损伤大鼠内质网应激的影响及其作用机制研究［J］.现代生物医学进展,2018,18(17):3245-3249.
WANG Y N,LI H J,YANG Y F,et al.Effects of adenosine receptor agonists on endoplasmic reticulum stress in rats with myocardial ischemia-reperfusion injury［J］.Progress Modern Biomed,2018,18(17):3245-3249.
［10］刘超,孟树萍,丁付燕,等.腺苷A2a受体激动剂(CGS21680)对体外循环大鼠心肌功能的影响及其作用机制［J］.中华实验外科杂志,2021,38(11):2195-2198.
LIU C,MENG S P,DING F Y,et al.Effects of adenosine A2a receptor agonist (CGS21680) on myocardial function in rats undergoing cardiopulmonary bypass and its mechanism［J］.Chin J Exp Surg,2021,38(11):2195-2198.
［11］　LONGA E Z,WEINSTEIN P R,CARLSON S,et a.Reversible middle cerebral artery occlusion without craniectomy in rats［J］.Stroke,1989,20(1):84-91．
［12］　赵欣,裴科阳,谭军.腺苷预处理对大鼠局灶性脑缺血再灌注后线粒体活性氧含量及线粒体呼吸链复合体Ⅰ、Ⅲ活性和脑组织总抗氧化能力的影响［J］.新乡医学院学报,2019,36(4):305-308.
ZHAO X,PEI K Y,TAN J.Effect of adenosine preconditioning on reactive oxygen species content,activity of respiratory chain complex Ⅰ and Ⅲ of mitochondria and the total antioxidant capacity of brain tissue after focal cerebral ischemia-reperfusion in rats［J］.J Xinxiang Med Univ,2019,36(4):305-308.
［13］　HEARSE D J.Reperfusion of the ischemic myocardium［J］.J Mol Cell Cardiol,1977,9(8):605-616.
［14］　阴雪妍,郭哨恺,谭军,等.腺苷预处理对大鼠脑缺血-再灌注损伤后脑内肿瘤坏死因子α和核因子kB表达的影响［J］.解剖学报,2020,51(6):839-847.
YIN X Y,GUO S K,TAN J,et al.Effcts of adenosine preconditioning on the expression of tumor necrosis factor α and nuclear factor kB in rats after cerebral ischemia-reperfusion injury［J］.Acta Anatomica Sinica,2020,51(6):839-847.
［15］　庞艳利,尉娜,谭军.腺苷预处理对大鼠脑缺血再灌注损伤的保护作用［J］.新乡医学院学报,2020,37(2):125-130.
PANG Y L,WEI N,TAN J.Protective effect of adenosine preconditioning on cerebral ischemia-reperfusion injury in rats［J］.J Xinxiang Med Univ,2020,37(2):125-130.
［16］　WANG L,ZHANG Z,WANG H.Naringin attenuates cerebral ischemia-reperfusion injury in rats by inhibiting endoplasmic reticulum stress［J］.Transl Neurosci,2021,12(1):190-197.
［17］　WU S,PIAO X,WANG N,et al.Naoluo Xintong capsule ameliorates apoptosis induced by endoplasmic reticulum stress in rats with cerebral ischemia/reperfusion injury［J］.Ann Palliat Med,2020,9(5):2913-2925.
［18］　ZHAO Y,FANG Y,ZHAO H,et al.Chrysophanol inhibits endoplasmic reticulum stress in cerebral ischemia and reperfusion mice［J］.Eur J Pharmacol,2018,818:1-9.
［19］　操良斌,桂勤芳,陈泳伊,等.浅低温联合硫化氢对全脑缺血再灌注后海马内质网应激的影响［J］.中华医学杂志,2018,98(44):3592-3597.
CAO L B,GUI Q F,CHEN Y Y,et al.Mild hypothermia combined with hydrogen sulfide alleviates hippocampal endoplasmic reticulum stress after global cerebral ischemia-reperfusion［J］.Natl Med J China,2018,98(44):3592-3597.
［20］　吴家鹏,李学智,汪莹,等.电针结合脑内注射血管内皮生长因子对脑缺血再灌注损伤大鼠内质网应激反应相关蛋白ATF6、IRE1、XBP1、CHOP的影响［J］.针刺研究,2018,43(6):341-346,359.WU J P,LI X Z,WANG Y,et al.Electroacupuncture combined with intracerebral injection of VEGF improves neurological dysfunction possibly by down-regulating expression of endoplasmic reticulum stress related proteins ATF6,etc.in cerebral ischemia-reperfusion injury rats［J］.Acupunct Res,2018,43(6):341-346,359.

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更新日期/Last Update: 2023-02-05