槲皮素对创伤性脑损伤大鼠血脑屏障保护作用的初步研究  

Preliminary study of protective effect of quercetin on blood-brain barrier in rats with traumatic brain injury

作　　者：秦荣 龚存林 张先斌 温剑峰[1] 张有昆 许海泽 邓磊[1] QIN Rong;GONG Cun-lin;ZHANG Xian-bin;WEN Jian-feng;ZHANG You-kun;XU Hai-ze;DENG Lei(The 908th Hospital of the Joint Logistics Support Force of the Chinese People’s Liberation Army,Yingtan 330002,Jiangxi,China)

机构地区：[1]中国人民解放军联勤保障部队第九〇八医院,江西鹰潭330002

出　　处：《生物医学工程与临床》2023年第2期131-137,共7页Biomedical Engineering and Clinical Medicine

基　　金：中国人民解放军联勤保障部队第九〇八医院专项课题(YGKTLC2016002)。

摘　　要：目的研究槲皮素(Que)治疗对创伤性脑损伤(TBI)大鼠血脑屏障的保护作用及其分子机制。方法选择SPF级雄性SD大鼠120只,鼠龄4~5个月,体质量约230~270 g。按随机数字法分为假手术组(SHAM组)、TBI组、Que治疗组(Que组),每组36只,12只备用。每组按实验后1 d、3 d、7 d分为3个亚组,每组12只。采用Feency重物自由落体撞击法建立TBI大鼠模型;SHAM组大鼠仅颅骨开骨窗,不进行撞针撞击;Que组大鼠在TBI建模后腹腔注射Que(100 mg/kg),每天1次,直至处死。分别于实验第1天、第3天、第7天对各组大鼠进行改良神经损伤严重程度评分(mNSS)评估大鼠神经功能损伤情况;干湿质量法检测脑组织含水量;采用伊文思蓝(EB)法检测各组大鼠血脑屏障的通透性;采用酶联免疫吸附分析(ELISA)法检测各组大鼠脑组织中白细胞介素(IL)-1β、肿瘤坏死因子α(TNF-α)、IL-10等炎性因子及基质金属蛋白酶9(MMP-9)的表达水平。结果与SHAM组比较,TBI组、Que组大鼠第1天、第3天、第7天的mNSS、脑组织内EB含量均明显增高(第1天:12.17分±0.93分、7.08分±0.79分vs 0.25分±0.45分;第3天:13.75分±0.75分、7.67分±0.78分vs 0.33分±0.49分;第7天:8.67分±0.65分、4.58分±0.67分vs 0.42分±0.51分。第1天:22.34μg/g±2.18μg/g、17.23μg/g±1.20μg/g vs 1.72μg/g±0.33μg/g;第3天:26.05μg/g±2.38μg/g、22.53μg/g±0.60μg/g vs 1.75μg/g±0.24μg/g;第7天:9.10μg/g±0.78μg/g、6.65μg/g±0.90μg/g vs 1.74μg/g±0.19μg/g)(P<0.05);Que组大鼠第1天、第3天、第7天的m NSS、脑组织含水量、脑组织内EB含量均较TBI组明显降低(P<0.05)。同时,大鼠脑组织含水量检测发现TBI组在第1天(79.73%±1.36%)、第3天(87.66%±1.20%)、第7天(81.21%±0.77%),Que组大鼠在第3天(78.23%±1.30%)均较SHAM组(73.53%±1.20%、73.43%±0.98%、73.65%±1.30%)升高(P<0.05)。ELISA检测结果发现,在第1天、第3天、第7天与SHAM组比较,TBI组、Que组大鼠脑组织中IL-1β、TNF-α、IL-10及MObjective To study the protective effect of quercetin(Que) on blood-brain barrier in rats with traumatic brain injury(TBI) and its molecular mechanism. Methods A total of 120 healthy male Sprague-Dawley rats were sacrificed, which aged4-5 months with body mass of 230-270 g. According to random number method, all of the rats were divided into sham operation group(SHAM group), TBI group and Que treatment group(Que group), with 36 cases in each group and 12 rats for standby. Each group was divided into 3 subgroups according to 1-day, 3-day and 7-day after experiment, 12 cases in each subgroup. The TBI model was established by Feency free fall impact method. In SHAM group, only the skull bone window was opened without impact. The rats in Que group were given intraperitoneally injection with Que(100 mg/kg) once a day after TBI modeling until sacrificed. The modified neurological severity score was performed on 1-day, 3-day and 7-day of experiment to evaluate neurological injury in each group. The water content in brain tissue was measured by dry wet mass method. Evans blue(EB) method was used to detect permeability of blood-brain barrier in each group. The expression levels of inflammatory factors such as interleukin(IL)-1β, tumor necrosis factor-α(TNF-α), IL-10 and matrix metalloproteinase-9(MMP-9) of brain tissue in each group were detected by enzyme-linked immunosorbent assay(ELISA). Results Compared with SHAM group, the modified neurological severity score(mNSS) and EB content in brain tissue of rats in TBI group and Que group were significantly increased on 1-day, 3-day and 7-day(1-day: 12.17 scores ± 0.93 scores, 7.08 scores ± 0.79 scores vs 0.25 scores ± 0.45scores;3-day: 13.75 scores ± 0.75 scores, 7.67 scores ± 0.78 scores vs 0.33 scores ± 0.49 scores;7-day: 8.67 scores ± 0.65 scores, 4.58scores ± 0.67 scores vs 0.42 scores ± 0.51 scores. 1-day: 22.34 μg/g ± 2.18 μg/g, 17.23 μg/g ± 1.20 μg/g vs 1.72 μg/g ± 0.33 μg/g;3-day:26.05 μg/g ± 2.38 μg/g, 22.53 μg/g ± 0.60 μg/g vs 1.75 μg/g

关 键 词：创伤性脑损伤 槲皮素 脑水肿 血脑屏障 炎性因子 基质金属蛋白酶9(MMP-9) 鼠 

分 类 号：R285.5[医药卫生—中药学] R-33[医药卫生—中医学] R651.15