【摘要】：目的：本研究建立新生SD大鼠缺氧缺血性腦損傷（hypoxic-ischemic brain damage，HIBD）模型，通過(guò)檢測(cè)不同時(shí)間點(diǎn)血清S100B蛋白水平，探討血清S100B蛋白的表達(dá)在新生SD大鼠缺氧缺血性腦病中的早期診斷價(jià)值，同時(shí)探索S100B蛋白在新生SD大鼠缺氧缺血性腦病后的出現(xiàn)時(shí)間、出現(xiàn)峰值時(shí)間及消失時(shí)間，從而進(jìn)一步為臨床新生兒缺氧缺血性腦病(HIE)患兒的早期診斷及確定HIE后S100B蛋白最佳采血時(shí)間提供實(shí)驗(yàn)依據(jù)。方法：清潔級(jí)，封閉群，健康新生7日齡SD大鼠168只，雌雄不限，體重12～18g，隨機(jī)分為3組：假手術(shù)組，HIBD模型組，，健康對(duì)照組，每組各56只，HIBD模型組采用改良Rice法制備HIBD模型。根據(jù)動(dòng)物處死時(shí)間不同HIBD組又隨機(jī)分為7個(gè)亞組：0h組、6h組、12h組、24h組、48h組、72h及96h組，每亞組各8只。假手術(shù)組及正常對(duì)照組也在相應(yīng)時(shí)間點(diǎn)分為7組，每組8只。各組大鼠在相應(yīng)的時(shí)間點(diǎn)斷頭取血后處死，收集血液及腦組織標(biāo)本。觀(guān)察各組運(yùn)動(dòng)行為學(xué)變化、腦組織形態(tài)學(xué)變化及HE染色；雙抗夾心ELISA法檢測(cè)血中S100B蛋白的變化。結(jié)果：(1)7日齡SD大鼠在HIBD組建模后出現(xiàn)不同程度的行為異常，正常對(duì)照組與假手術(shù)組鼠未見(jiàn)異常行為(2)正常對(duì)照組、假手術(shù)組腦組織外觀(guān)正常，兩側(cè)對(duì)稱(chēng)，無(wú)水腫、萎縮及壞死；HIBD模型組在建模后各時(shí)間點(diǎn)肉眼觀(guān)腦組織大體形態(tài)出現(xiàn)不同程度的異常改變(3)HE染色：正常對(duì)照組、假手術(shù)組腦組織和HIBD模型組在建模后0h腦細(xì)胞層次清楚、細(xì)胞形態(tài)正常；HIBD模型組在建模后6h出現(xiàn)不同程度的神經(jīng)細(xì)胞水腫、變性；12h、24h腦組織出現(xiàn)淤血水腫和血管充血；48h、72h神經(jīng)細(xì)胞出現(xiàn)不同程度的壞死，細(xì)胞核碎裂、溶解；96h細(xì)胞出現(xiàn)大片狀，細(xì)胞核碎裂、溶解。(4)血清中S100B蛋白的表達(dá)：HIBD組0h時(shí)間點(diǎn)組與正常對(duì)照組、假手術(shù)組比較血清中S100B蛋白差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）；HIBD組（0h、96h時(shí)間點(diǎn)組除外）、與正常對(duì)照組、假手術(shù)組比較各個(gè)時(shí)間點(diǎn)血清中S100B蛋白值顯著升高（P0.05），差異有統(tǒng)計(jì)學(xué)意義；HIBD組48h時(shí)間點(diǎn)組分別與HIBD組6h、12h、24h、72h時(shí)間點(diǎn)組比較血清中S100B蛋白值顯著升高（P0.05），差異有統(tǒng)計(jì)學(xué)意義；HIBD組6h、12h、24h、72h時(shí)間點(diǎn)組組間比較血清中S100B蛋白值差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。正常對(duì)照組、假手術(shù)組、HIBD組血清中S100B蛋白含量在0h為18.89±4.68ng/L、15.21±3.27ng/L、14.39±5.68ng/L；6h分別為17.36±1.24ng/L、15.77±1.59ng/L、27.61±6.30ng/L；12h分別為16.32±2.33ng/L、15.39±0.96ng/L、21.93±5.53ng/L；24h分別為17.01±1.66ng/L、15.12±1.10ng/L、24.65±4.83ng/L；48h分別為18.33±1.99ng/L、15.45±1.78ng/L、44.25±4.12ng/L；72h分別為18.47±1.85ng/L、15.21±1.94ng/L、28.39±4.53ng/L；96h分別為17.61±2.57ng/L、15.40±0.99ng/L、17.01±2.56ng/L結(jié)論：（1）新生大鼠缺氧缺血性腦病模型建立成功。（2）S100B蛋白可作為新生大鼠缺氧缺血性腦病早期診斷的生化指標(biāo)。（3）新生大鼠缺氧缺血性腦病血清中S100B蛋白在缺氧缺血發(fā)生后6小時(shí)開(kāi)始升高，48小時(shí)達(dá)到峰值，72小時(shí)開(kāi)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.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R722.1

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