新生大鼠缺氧缺血性脑病血清S100B蛋白表达变化的实验研究

发布时间：2018-08-12 20:17

【摘要】：目的：本研究建立新生SD大鼠缺氧缺血性脑损伤（hypoxic-ischemic brain damage，HIBD）模型，通过检测不同时间点血清S100B蛋白水平，探讨血清S100B蛋白的表达在新生SD大鼠缺氧缺血性脑病中的早期诊断价值，同时探索S100B蛋白在新生SD大鼠缺氧缺血性脑病后的出现时间、出现峰值时间及消失时间，从而进一步为临床新生儿缺氧缺血性脑病(HIE)患儿的早期诊断及确定HIE后S100B蛋白最佳采血时间提供实验依据。方法：清洁级，封闭群，健康新生7日龄SD大鼠168只，雌雄不限，体重12～18g，随机分为3组：假手术组，HIBD模型组，，健康对照组，每组各56只，HIBD模型组采用改良Rice法制备HIBD模型。根据动物处死时间不同HIBD组又随机分为7个亚组：0h组、6h组、12h组、24h组、48h组、72h及96h组，每亚组各8只。假手术组及正常对照组也在相应时间点分为7组，每组8只。各组大鼠在相应的时间点断头取血后处死，收集血液及脑组织标本。观察各组运动行为学变化、脑组织形态学变化及HE染色；双抗夹心ELISA法检测血中S100B蛋白的变化。结果：(1)7日龄SD大鼠在HIBD组建模后出现不同程度的行为异常，正常对照组与假手术组鼠未见异常行为(2)正常对照组、假手术组脑组织外观正常，两侧对称，无水肿、萎缩及坏死；HIBD模型组在建模后各时间点肉眼观脑组织大体形态出现不同程度的异常改变(3)HE染色：正常对照组、假手术组脑组织和HIBD模型组在建模后0h脑细胞层次清楚、细胞形态正常；HIBD模型组在建模后6h出现不同程度的神经细胞水肿、变性；12h、24h脑组织出现淤血水肿和血管充血；48h、72h神经细胞出现不同程度的坏死，细胞核碎裂、溶解；96h细胞出现大片状，细胞核碎裂、溶解。(4)血清中S100B蛋白的表达：HIBD组0h时间点组与正常对照组、假手术组比较血清中S100B蛋白差异无统计学意义（P0.05）；HIBD组（0h、96h时间点组除外）、与正常对照组、假手术组比较各个时间点血清中S100B蛋白值显著升高（P0.05），差异有统计学意义；HIBD组48h时间点组分别与HIBD组6h、12h、24h、72h时间点组比较血清中S100B蛋白值显著升高（P0.05），差异有统计学意义；HIBD组6h、12h、24h、72h时间点组组间比较血清中S100B蛋白值差异无统计学意义（P0.05）。正常对照组、假手术组、HIBD组血清中S100B蛋白含量在0h为18.89±4.68ng/L、15.21±3.27ng/L、14.39±5.68ng/L；6h分别为17.36±1.24ng/L、15.77±1.59ng/L、27.61±6.30ng/L；12h分别为16.32±2.33ng/L、15.39±0.96ng/L、21.93±5.53ng/L；24h分别为17.01±1.66ng/L、15.12±1.10ng/L、24.65±4.83ng/L；48h分别为18.33±1.99ng/L、15.45±1.78ng/L、44.25±4.12ng/L；72h分别为18.47±1.85ng/L、15.21±1.94ng/L、28.39±4.53ng/L；96h分别为17.61±2.57ng/L、15.40±0.99ng/L、17.01±2.56ng/L结论：（1）新生大鼠缺氧缺血性脑病模型建立成功。（2）S100B蛋白可作为新生大鼠缺氧缺血性脑病早期诊断的生化指标。（3）新生大鼠缺氧缺血性脑病血清中S100B蛋白在缺氧缺血发生后6小时开始升高，48小时达到峰值，72小时开始下降。
[Abstract]:AIM: To establish a hypoxic-ischemic brain damage (HIBD) model in neonatal SD rats. To investigate the early diagnostic value of serum S100B protein expression in neonatal SD rats with hypoxic-ischemic encephalopathy (HIBD) by detecting the levels of serum S100B protein at different time points. The occurrence time, peak time and disappearance time of ischemic encephalopathy provide experimental basis for early diagnosis of clinical neonatal hypoxic-ischemic encephalopathy (HIE) and determination of the best time for collecting S100B protein after HIE. Methods: 168 healthy neonatal SD rats of 7 days old, male or female, weighing 12-1. 8 g, randomly divided into three groups: sham operation group, HIBD model group, healthy control group, 56 in each group, HIBD model group using modified Rice method HIBD model group. According to the different time of animal execution, the HIBD model group was randomly divided into seven subgroups: 0 h group, 6 h group, 12 h group, 24 h group, 48 h group, 72 h and 96 h group, each subgroup had 8 rats. The rats in each group were sacrificed at the corresponding time points and blood and brain tissue samples were collected. The changes of motor behavior, brain morphology and HE staining were observed. The changes of S100B protein in blood were detected by sandwich ELISA. The normal control group and sham operation group showed no abnormal behavior (2) normal control group, sham operation group showed normal appearance of brain tissue, bilateral symmetry, no edema, atrophy and necrosis; HIBD model group showed abnormal changes in the gross morphology of brain tissue at various time points after modeling (3) HE staining: normal control group In the sham operation group, the brain tissue and the HIBD model group had clear level of brain cells and normal cell morphology at 0 h after modeling; in the HIBD model group, there were different degrees of neuronal edema and degeneration at 6 h after modeling; in the 12 h and 24 h brain tissues, there were congestion and congestion; in the 48 h and 72 h brain cells appeared different degrees of necrosis, nuclear fragmentation and lysis; and in the 96 h after modeling, there were no significant differences in neuronal degeneration. The expression of S100B protein in serum of HIBD group at 0 h time point was not significantly different from that of normal control group (P 0.05), HIBD group (except for 0 h and 96 h time point group), sham operation group and normal control group. The serum levels of S100B protein in HIBD group were significantly higher than those in HIBD group at 6 h, 12 h, 24 h and 72 h time points (P 0.05), and there was significant difference between HIBD group at 6 h, 12 h, 24 h and 72 h time points (P 0.05). The control group, sham-operation group, HIBDgroup, HIBDgroup, sham-operation group, HIBDgroup, HIBDgroup, the serum S100B protein content in 0 h was 18.89 +4.68ng/L, 15.21 [(3.27ng/L), 15.21 [(3.27ng/L), 14.39 [(5.68 ng/L), 14.39 [(14.39 [5.68 ng/L), 17.36 [(1.36 [1.24ng/L), 15.77 [1.24ng/L, 15.77 [1.59ng/L, 15.77 [1.59 ng/L, 27.61 [6.61 [6.61 [6.30 ng/L]]/L, 16.32 [L, 24.65 + 4.83ng/L; 48H Conclusion: (1) The establishment of hypoxic-ischemic encephalopathy model in neonatal rats was successful. (2) S100B protein can be used as hypoxic-ischemic encephalopathy model in neonatal rats. Biochemical markers for early diagnosis of hypoxic-ischemic encephalopathy. (3) Serum S100B protein in neonatal rats began to increase 6 hours after hypoxic-ischemic encephalopathy, peaked at 48 hours, and began to decrease at 72 hours.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R722.1

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