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κB表达的影响

doi:10.16098/j.issn.0529-1356.2020.06.006

摘要/Abstract

摘要：

目的通过观察腺苷预处理对脑缺血-再灌注（IR）损伤后大鼠脑内肿瘤坏死因子α（TNF-α）和核因子κB（NF-κB）表达的影响，探讨腺苷对大鼠脑缺血-再灌注损伤的保护机制。方法 Sprague-Dawley大鼠60只随机分为假手术组（F组）、缺血-再灌注组（IR组）和腺苷预处理缺血-再灌注组（AP组），每组动物按照缺血-再灌注后2 h、6 h、24 h及48 h这4个时间点随机分组，每组5只。采用大脑中动脉阻塞（MCAO）法制备大鼠大脑中动脉闭塞模型。通过对以上3组脑缺血再灌注48 h后的大鼠脑组织行头颅MRI、HE染色以及对以上60只大鼠进行神经功能缺损评分，通过免疫组织化学方法观察大鼠脑组织中TNF-α和NF-κB的平均积分吸光度值进一步验证腺苷预处理对大鼠脑缺血-再灌注损伤的脑保护作用，探讨腺苷对大鼠脑缺血-再灌注损伤的保护机制。结果 F组大鼠术后无神经功能缺损体征，AP组和IR组大鼠术后均出现明显神经功能缺损体征。AP组和IR组大鼠神经功能缺损评分显著高于F组，AP组大鼠神经功能缺损评分显著低于IR组，差异均具有统计学意义（P＜0.05）。F组大鼠术后MRI中T1WI、T2WI均未见明显异常，AP组和IR组大鼠脑梗死体积显著高于F组，AP组大鼠脑梗死体积显著低于IR组，差异均具有统计学意义（P＜0.05）。AP组和IR组术后大鼠脑组织中TNF-α和NF-κB的表达显著高于F组，AP组术后大鼠脑组织中TNF-α和NF-κB的表达显著低于IR组，差异均具有统计学意义（P＜0.05）。结论腺苷预处理可通过减少大鼠缺血-再灌注损伤后脑内TNF-α和NF-κB的表达而减轻神经细胞损伤，达到其脑保护作用。

关键词: 腺苷, 预处理, 缺血-再灌注, 肿瘤坏死因子α, 核因子κB, 免疫组织化学, 大鼠

Abstract:

Objective To observe the effect of adenosine preconditioning on the expression of tumor necrosis factor α（TNF-α）and nuclear factor κB (NF-κB)in rats after cerebral ischemia-reperfusion(IR) injury,and to explore the protective mechanism of adenosine on focal cerebral ischemia-reperfusion injury in rats. Methods Total sixty Sprague-Dawley rats were randomly divided into sham group (F group),ischemia-reperfusion group (IR group) and adenosine preconditioning ischemia-reperfusion group (AP group). Each group was randomly divided into 5 mice at 2 hour, 6 hour, 24 hour and 48 hour after ischemia-reperfusion. The rat moadel of middle cerebral artery occlusion (MCAO) was established by suppository method . 48 hours after ischemia-reperfusion, line head MRI, HE staining and for more than of 60 neursl function defect scale in rats，integral optical density of the software system by immunohistochemical method in the observation group rats TNF-α and NF-κB average absorbance values of rats’brain tissues to further verify the adenosine preconditioning on rat cerebral ischemia-reperfusion injury，adenosine on rat focal cerebral ischemia-reperfusion injury of protection mechanism. Results No signs of neurological defects was found in the rats in F after surgery, and obvious signs of neurological defects were found in the rats of AP and IR groups.The neurological defect scores of the AP group and the IR group were significantly higher than those of the F group, and the neurological defect scores of the AP group were significantly lower than those of the IR group, with statistically significant differences(P<0.05).Postoperative MRI images of T1WI and T2WI of rats in group F showed no obvious abnormalities. The cerebral infarction volume of rats in the AP group and the IR group was significantly higher than those of the F group, and the cerebral infarction volume of rats in the AP group was significantly lower than those of the IR group,with statistically significant differences(P<0.05).The expressions of TNF-α and NF-κB in the brain tissues of the AP group and the IR group were significantly higher than those of the F group, and the expressions of TNF-α and NF-κB in the brain tissues of the AP group was significantly lower than those of the IR group, with statistically significant differences(P<0.05). Conclusion Adenosine preconditioning can protect the brain by descreasing the expression of TNF-α and NF-κB in rats with cerebral ischemia-reperfusion injury.

Key words: Adenosine, Pretreatment, Ischemia-reperfusion, Tumor necrosis factor α, Nuclear factor κB, Immunohistochemistry, Rat

中图分类号:

R743.3

阴雪妍郭哨恺谭军范文统李明. 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κB表达的影响[J]. 解剖学报, 2020, 51(6): 839-847.

YIN Xue-yan GUO Shao-kai TAN Jun FAN Wen-tong LI Ming. Effcts of adenosine preconditioning on the expression of tumor necrosis factor α and nuclear factor κB in rats after cerebral ischemia-reperfusion injury[J]. Acta Anatomica Sinica, 2020, 51(6): 839-847.

参考文献

［1］ Zhang W, Fan YW, Gao J, et al. Review of relevant literature on epidemiological investigation of stroke ［J］. Chinese Clinical Neuroscience, 2014,22 (6):699-703.(in Chinese)

张薇,范宇威,高静,等.脑卒中流行病学调查相关文献复习［J］.中国临床神经科学,2014,22（6）：699-703.

[2］ Liu L, Liu LH, Ma YK. Research progress in the mechanism of cerebral ischemia-reperfusioninjury ［J］. Pharmaceutical Research,2016-35 (9):542-544. (in Chinese)

刘磊,刘丽华,马玉奎.脑缺血再灌注损伤机制研究进展［J］.药学研究,2016-35（9）：542-544.

［3］ Dong MZh, Deng Y, Liu F, et al. Research progress in the mechanism of cerebral ischemia-reperfusion injury ［J］. Grass-Roots Medical Forum, 2018,22 (14):1981-1982. (in Chinese)

董梦珍,邓云,刘飞,等.脑缺血再灌注损伤机制研究进展［J］.基层医学论坛,2018,22（14）：1981-1982.

［4］ Zhang BY, Ma LL, Zhu L, et al. Research status of autophagy in ischemia reperfusioninjury ［J］. World Latest Medical Information Digest,2017,17 (26):65-66. (in Chinese)

张冰缘,马丽丽,朱琳,等.自噬在缺血再灌注损伤中的研究现状［J］.世界最新医学信息文摘,2017,17（26）：65-66.

［5］ Huang TT, Teng L, Huang H, et al. Effects of butylphthalide pretreatment on neurological deficit score, oxidative damage and morphology in rats with cerebral ischemia-reperfusion injury ［J］. Acta Anatomica Sinica, 2014, 45 (5):622-626. (in Chinese)

黄婷婷,滕蕾,黄惠,等，丁苯酞预处理对脑缺血再灌注损伤大鼠神经功能缺损评分、氧化损伤和形态学的影响［J］.解剖学报，2014,45（5）：622-626.

[6］ Eitzschig HK, Sitkovsky MV, Robson SC. Purinergic signaling during inflammation ［J］.N Eng J Med, 2013,368(13):1260.

［7］ Ohta A, Sitkovsky M. Role of protein-coupled adenosine receptors in downregulation of inflammation and peotection from tissue damage ［J］.Nature,2001,414(6866):916-920.

［8］ Yu N, Tan J, Lu T. Protective effect and mechanism of adenosine preconditioning on ischemia reperfusion injury in rats ［J］. Chinese Journal of Applied Neurological Diseases,2011,14 (3): 1-3. (in Chinese)

尉娜,谭军,路坦.腺苷预处理对大鼠缺血再灌注损伤的保护作用和机制［J］.中国实用神经疾病杂志,2011,14（3）：1-3.

［9］ Tao X, Kuang PG, Ming J, et al. Protective effects of adenosine on lipid peroxidation during ischemia-reperfusion injury ［J］. Journal of Brain and Nervous Diseases, 1996, 4 (1):1-3. (in Chinese)

陶昕,匡培根,名晶，等.腺苷对缺血再灌注损伤时的保护作用——抗脂质过氧化［J］.脑与神经疾病杂志,1996，4（1）：1-3.

［10］ Tan J, Sun ZhY. Effects of adenosine preconditioning on glial neurotrophic factor expression after focal cerebral ischemia-reperfusion injury in rats ［J］. Chinese Journal of Practical Medicine, 2008,35 (21): 1-3. (in Chinese)

谭军,孙兆印.腺苷预处理对大鼠局灶性脑缺血再灌注损伤后胶质源性神经营养因子表达的影响［J］.中国实用医刊，2008，35（21）:1-3.

［11］ Zhang Y, Chen J, Han YY, et al. Effects of adenosine preconditioning on hypoxia-inducer-1 expression in brain tissue of rats with focal cerebral ischemia-reperfusion injury ［J］. Journal of Xinxiang Medical College, 2014, 31(4): 272-274. (in Chinese)

张妤,陈娟,韩艳艳,等.腺苷预处理对局灶性脑缺血再灌注损伤大鼠脑组织中低氧诱导因子-1α表达的影响［J］.新乡医学院校报，2014,31(4)：272-274．

［12］ Yu N, Wang JP, Shen XL, et al. Adenosine preconditioning on VEGF expression after cerebral ischemia-reperfusion injury in rats ［J］. Journal of Stroke and Neurological Disease, 2014, 31 (2):107-109. (in Chinese)

尉娜,王建平,申小龙,等.腺苷预处理对大鼠脑缺血再灌注损伤后VEGF的表达［J］.中风与神经疾病杂志，2014,31（2）：107-109.

［13］ Wang J, Han YY, Li J, et al. Changes of CX3CL1 and glt-1 expression in astrocytes after adenosine pretreatment with oxyglucose deprivation reoxygenation ［J］. Chinese Journal of Geratology,2017, 37(2):288-291. (in Chinese)

王洁,韩艳艳,李娟,等.腺苷预处理星形胶质细胞氧糖剥夺复氧糖后CX3CL1、GLT-1表达的变化［J］.中国老年学杂志,2017,37(2):288-291.

［14］ Niu CY, Hu YN,Wang J, et al. Effects of adenosine preconditioning on expression of astrocytes and lipid carrier protein-2 induced by glucose deprivation/reoxygenation ［J］. Chinese Journal of Anatomy, 2016,39(2):192-195. (in Chinese)

牛草原,胡亚男,王洁,等.腺苷预处理对氧糖剥夺/复氧糖诱导的星形胶质细胞及脂质运载蛋白2表达的影响［J］.解剖学杂志,2016,39(2):192-195.

［15］ Zhao X, Pei KY, Tan J. Adenosine preconditioning on focal cerebral ischemia reperfusion in rats after mitochondrial reactive oxygen content and mitochondrial respiratory chain complexes Ⅰ, Ⅲ activity of brain tissue and total antioxidant capacity influence ［J］. Journal of Xinxiang Medical College, 2019, 36(4): 305-308. (in Chinese)

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.

［16］ Pei KY, Tan J, Xia YH, et al. Adenosine preconditioning on mitochondrial function of nerve cells in rats with focal cerebral ischemia-reperfusion injury ［J］. Shandong Medical Journal, 2008,58(27):43-46. (in Chinese)

裴科阳,谭军,夏艳红,等.腺苷预处理的局灶性脑缺血再灌注损伤大鼠神经细胞线粒体功能观察［J］.山东医药,2018,58(27):43-46.

［17］ Yu N, Zhao JH, Li J, et al. Effects of adenosine preconditioning on Wnt1 expression in rats with focal cerebral ischemia-reperfusion injury ［J］. Journal of Stroke and Neurological Disease, 2015,32(8):691-694. (in Chinese)

尉娜,赵建华,李娟,等.腺苷预处理对局灶性脑缺血再灌注损伤大鼠脑内Wnt1表达的影响［J］.中风与神经疾病杂志,2015,32(8):691-694.

［18］ Read MA, Whitley MZ ,Williams AJ, et al.NF-Kappa B and I kappa Bα：an inducible regulatory system in endothelial activation［J］.J Exp Med,1994,179（2）:503-512.

［19］ Feng ChSh, Wang HSh, Li S. Expression of nuclear transcription factor B after transient focal cerebral ischemia/reperfusion in rats ［J］. Journal of Stroke and Neurological Diseases, 2003,20(1):11-13. (in Chinese)

冯春生,王虎山,李淞.大鼠短暂局灶性脑缺血再灌注后核转录因子Κb的表达［J］.中风与神经疾病杂志,2003,20(1):11-13.

［20］ Zhang DM,Liu JX, Chen HB, et al. Expression of NF- B and il-6 in the brain of rats with focal cerebral ischemia-reperfusion injury ［J］. Chinese Journal of Geratology, 2005, 25 (11): 1364-1366. (in Chinese)

张冬梅,刘举祥,陈红兵,等.NF-κB和IL-6在局灶性脑缺血再灌注损伤大鼠脑组织中的表达［J］.中国老年学杂志,2005,25(11):1364-1366.

［21］ Mecocci P, Mariani E, Polidori MC, et al. Antioxidant agents in Alzheimer’s disease［J］.Cent Nerv Syst Agents Med Chem,2008,8(1):48-63.

［22］ Li YSh, Ding SJ, Ni CR. Effects of nuclear transcription factor -B on apoptosis of rat nerve cells after cerebral ischemia-reperfusion and its mechanism ［J］. Practical Journal of Medicine & Pharmacy, 2005,22(8):706-709. (in Chinese)

李永生,丁素菊,倪灿荣.核转录因子-κB对鼠脑缺血再灌注后神经细胞凋亡的影响及机制［J］.实用医药杂志,2005,22(8):706-709.

［23］ Taylor CT,Dzus Aland ,Colgan SP. Autocrine regulation of epithelial permeability by hypoxia：role for polarized release of tumor necrosis factor alpha［J］. Gastroenterology, 1998, 114(4):657-668.

［24］ Trickler WJ, Mayhan WG, Miller DW. Brain microvessel endothelial cell responses to tumor necrosis factor-alpha involve a nuclear factor kappaB （NF-kappaB）signal transduction pathway［J］.Brain Res, 2005,1048(1-2):24-31.

［25］ Li JJ, Jiang HY, Shao JL. Expression of tnf-a, il-6, il-1 in brain tissue of rats with cerebral ischemia-reperfusion injury ［J］. Journal of Kunming Medical University, 2016,37 (9):31-35. (in Chinese)

李俊杰,蒋海燕,邵建林.脑缺血-再灌注损伤大鼠脑组织中TNF-a,IL-6,IL-1β的表达［J］.昆明医科大学学报,2016,37（9）:31-35.

[1]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[2]	郑丽平刘霞杜晓华吴亚娟刘珊珊 . 脑源性神经营养因子在牦牛与黄牛端脑不同区域的表达与分布 [J]. 解剖学报, 2024, 55(1): 10-16.
[3]	刘丽平朱梦妮徐慧 . 白细胞介素6对脂多糖诱导所致类子痫前期大鼠有核红细胞的影响 [J]. 解剖学报, 2023, 54(6): 722-729.
[4]	梁盼盼禹爱梅杜静寇文辉王欢欢宋爱霞. 基于c-Jun氨基末端激酶/p38丝裂原活化蛋白激酶信号通路探讨阿魏酸钠对偏头痛大鼠炎症反应的抑制作用[J]. 解剖学报, 2023, 54(6): 652-659.
[5]	姚素华鄢骏谢芳林春华黄争春 . 奥氮平介导环磷酸腺苷反应元件结合蛋白/脑源性神经营养因子/酪氨酸蛋白激酶受体B通路对精神分裂模型大鼠的神经修复作用 [J]. 解剖学报, 2023, 54(6): 660-667.
[6]	刘叶楚亚楠徐岑璐何佳澄苏炳银太颢然. Roscovitine挽救帕金森病小鼠相关脑区神经元丢失和神经炎症[J]. 解剖学报, 2023, 54(6): 635-643.
[7]	阎锟武筱林刘英娟葛科立任雷鸣李红云 . 脑蛋白水解物-Ⅰ对帕金森病小鼠肠道菌群的调节作用[J]. 解剖学报, 2023, 54(5): 497-504.
[8]	孙娟陈敬威杨艺林涛牟雅琳赵秀丽曾国熙张艳 . 缺氧预处理通过缺氧诱导因子-1α/基质细胞衍生因子-1通路对大鼠血液学相关指标的影响[J]. 解剖学报, 2023, 54(5): 505-511.
[9]	燕洋洋郭凯孙志宏 . 嘌呤能离子通道型受体7介导的帕金森大鼠认知功能障碍[J]. 解剖学报, 2023, 54(5): 531-537.
[10]	庄海容吴子东陈雪红李成军. 舒洛地特对大鼠糖尿病视网膜病变的修复及MAPK通路的调节作用[J]. 解剖学报, 2023, 54(4): 392-399.
[11]	于华婧魏路阳刘姗姗管成剑张忠涛. MLLT1超伸长复合亚基在肝细胞癌发生发展中的作用及其临床意义[J]. 解剖学报, 2023, 54(4): 425-433.
[12]	王潇李梁崔成立蔡志平于畅张立佳 . 耐力运动联合间歇性冷刺激促进大鼠白色脂肪组织棕色化[J]. 解剖学报, 2023, 54(3): 348-356.
[13]	张沂洲李莎米世雄左洪春张倩倩李晗琳雷梓晗张东泽崔慧先. 雌二醇经雌激素受体调控分拣蛋白相关受体A对小鼠海马神经元树突棘密度和突触蛋白表达的影响[J]. 解剖学报, 2023, 54(3): 261-268.
[14]	刘柯婷陈缤彬税玉莲石清明肖莉. 外源性H2S对帕金森病模型小鼠神经保护作用及其对基质金属蛋白酶9和Caspase-1表达的影响[J]. 解剖学报, 2023, 54(3): 289-295.
[15]	侯翌葳杨宇王博段英涛姚宏波. 普兰林肽对5×FAD小鼠和WT小鼠认知行为及脑和视网膜β淀粉样蛋白6E10、炎症因子和神经细胞形态的影响[J]. 解剖学报, 2023, 54(3): 283-288.