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丰富环境预处理对脑缺血再灌注损伤大鼠肿瘤坏死因子α和白细胞介素1β表达及细胞凋亡的影响
高振坤1,2,申鑫雅2,韩宇2,郭祎莎2,袁美2,毕霞2*
0
(1. 上海中医药大学附属曙光医院康复医学科, 上海 201203;
2. 上海健康医学院附属周浦医院康复医学科, 上海 201318
*通信作者)
摘要:
目的 探讨丰富环境预处理对脑缺血再灌注后大鼠梗死周围皮质TNF-α和IL-1β表达及细胞凋亡的影响。方法 将45只雄性SD大鼠随机分为假手术组、模型组和丰富环境预处理组,每组15只。造模前,假手术组和模型组大鼠置于标准环境笼中饲养,丰富环境预处理组大鼠置于丰富环境笼中饲养,均持续3周。模型组和丰富环境预处理组大鼠均采用改良Longa线栓法建立短暂性大脑中动脉栓塞模型,假手术组大鼠不插入线栓。术后1~3 d采用改良神经功能缺损评分(mNSS)评估大鼠的神经功能。术后3 d取脑组织,通过H-E染色观察梗死周围皮质组织病理学变化,TUNEL检测观察细胞凋亡情况,免疫荧光检测观察小胶质细胞离子钙结合适配器分子1(Iba-1)激活情况,蛋白质印迹法检测TNF-α和IL-1β表达情况。结果 与假手术组相比,模型组和丰富环境预处理组大鼠mNSS升高(P均<0.05),梗死周围皮质出现明显病理损伤,TUNEL阳性细胞数量增加(P<0.05),小胶质细胞Iba-1激活增多(P均<0.05),TNF-α和IL-1β蛋白表达水平升高(P均<0.05)。与模型组相比,丰富环境预处理组大鼠mNSS降低(P<0.05),梗死周围皮质病理损伤减轻,TUNEL阳性细胞数量降低(P<0.05),小胶质细胞Iba-1激活减少(P<0.05),TNF-α和IL-1β蛋白表达水平下降(P均<0.05)。结论 丰富环境预处理可减轻大鼠脑缺血再灌注后的神经功能障碍和梗死周围脑组织损伤,其机制可能与抑制梗死周围皮质的炎症反应和细胞凋亡有关。
关键词:  脑缺血再灌注  预处理  丰富环境  炎症  细胞凋亡
DOI:10.16781/j.CN31-2187/R.20220533
投稿时间:2022-06-22修订日期:2022-09-02
基金项目:上海市浦东新区卫生健康委员会临床特色学科建设项目(PWYts2021-10).
Effects of enriched environment preconditioning on expression of tumor necrosis factor α and interleukin 1β and apoptosis in rats after cerebral ischemia-reperfusion injury
GAO Zhenkun1,2,SHEN Xinya2,HAN Yu2,GUO Yisha2,YUAN Mei2,BI Xia2*
(1. Department of Rehabilitation Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;
2. Department of Rehabilitation Medicine, Zhoupu Hospital Affiliated to Shanghai University of Medicine & Health Sciences, Shanghai 201318, China
*Corresponding author)
Abstract:
Objective To explore the effects of enriched environment preconditioning on the expression of tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) and apoptosis in the peri-infarct cortex of rats after cerebral ischemia-reperfusion. Methods A total of 45 male Sprague-Dawley rats were randomly divided into sham group, model group and enriched environment preconditioning group, with 15 rats in each group. Before modeling, rats in the sham group and model group were fed in standard environment cages, while rats in the enriched environment preconditioning group were fed in enriched environment cages, all for 3 weeks. The transient middle cerebral artery occlusion model was established by modified Longa's method in the model group and enriched environment preconditioning group, while rats in the sham group were not inserted with suture. Modified neurological severity score (mNSS) was used to evaluate the neurological function 1-3 d after operation. The brain tissue was harvested 3 d after the operation, the pathological changes of the peri-infarct cortex were observed by hematoxylin-eosin staining, the apoptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL), the activation of microglia ionized calcium-binding adaptor molecule 1 (Iba-1) was observed by immunofluorescence, and the expression of TNF-α and IL-1β was detected by Western blotting. Results Compared with the sham group, the peri-infarct cortical tissue in the rats of model and enriched environment preconditioning groups had obvious pathological changes, and the mNSS, TUNEL positive cell number, the activation of microglia Iba-1, the expression of TNF-α and IL-1β were increased (all P<0.05). Compared with the model group, mNSS, TUNEL positive cell number, the activation of microglia Iba-1, and the expression of TNF-α and IL-1β were decreased in the enriched environment preconditioning group (all P<0.05) and the degree of peri-infarct cortical tissue injury was alleviated. Conclusion Enriched environment preconditioning can attenuate neurological function and peri-infarct brain tissue injuries in rats after cerebral ischemia-reperfusion, and the mechanism may be related to the inhibition of inflammation and apoptosis in the peri-infarct cortex.
Key words:  cerebral ischemia-reperfusion  preconditioning  enriched environment  inflammation  apoptosis