高胆红素血症及胆红素脑病新生大鼠模型的建立及评价

发布时间：2018-06-15 23:19

本文选题：高胆红素血症 + 胆红素脑病　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:建立可行可靠、步骤简单且具有良好模拟性的高胆红素血症及胆红素脑病的7日龄SD大鼠模型,为在临床实践中探究高胆红素血症及胆红素脑病的发病机制及并发症的防治提供模型基础。方法:选取7日龄的SD(Sprague-Dawley,SD)大鼠50只随机分成5组,分别给予生理盐水,50mg/kg、100mg/kg、150mg/kg、200mg/kg晶体胆红素腹腔内注射。在实验处理24h后取大鼠脑组织及心脏取血,测定血清、脑组织的胆红素水平及S-100β浓度,进行脑组织HE染色观察海马区神经细胞的病理改变,用TUNEL法测定神经细胞凋亡数,计算凋亡率(Apoptotic index,AI)并及时记录各组各指标的实验结果,计算相关数据并对其进行统计学分析,得出结论。进一步评价新生大鼠模型的可靠性及稳定性。结果:(1)观察实验动物一般情况:实验前,各组大鼠一般情况良好,皮肤颜色红润,弹性好,对外界刺激反应灵敏,活动正常。干预24小时后观察:生理盐水组的大鼠皮肤颜色红润,对外界刺激反应灵敏;50mg/kg组的大鼠皮肤欠红润,弹性较差,对外界刺激反应迟缓;100mg/kg组与150 mg/kg组新生大鼠的皮肤颜色黄染,弹性差,对外界刺激反应迟钝,有肌张力降低所致的翻滚及俯伏;200 mg/kg组的大鼠皮肤呈黄绿色,弹性极差,对外界刺激无反应甚至昏迷。(2)HE染色:生理盐水组海马区神经细胞排列整齐,结构完整,形态正常;50mg/kg组可见神经细胞之间轻度的肿胀、变性;100mg/kg组与150mg/kg组神经元结构紊乱,细胞之间肿胀、变形明显,部分细胞核染色质致密浓缩,胞浆空化;200mg/kg组神经细胞之间极度肿胀、变形,可见多数细胞核染色质致密浓缩,胞浆空化。(3)血清及脑组织胆红素水平及S-100β浓度:与生理盐水组相比,各组的血清及脑组织胆红素水平及S-100β浓度均有统计学差异(P0.05);与50mg/kg组相比,100mg/kg组血清及脑组织的S-100β无显著性差异(P0.05)。(4)TUNEL法测凋亡指数(AI):与生理盐水组相比,各组的凋亡指数均有统计学差异(P0.05);与150mg/kg组相比,200mg/kg组凋亡指数无显著性差异(P0.05)。结论:(1)7d SD大鼠腹腔注射50 mg/kg晶体胆红素可建立高胆红素血症动物模型。(2)7d SD大鼠腹腔注射100mg/kg与150 mg/kg晶体胆红素可建立稳定的胆红素脑病模型。(3)当腹腔注射150 mg/kg晶体胆红素时,神经胶质细胞分解明显,所以7d SD大鼠注射150 mg/kg晶体胆红素可建立胶质细胞损伤指标的模型。
[Abstract]:Objective: to establish a 7 day old SD rat model of hyperbilirubinemia and bilirubin encephalopathy. To provide a model for exploring the pathogenesis of hyperbilirubinemia and bilirubin encephalopathy and the prevention and treatment of complications in clinical practice. Methods: fifty SD rats aged 7 days were randomly divided into 5 groups, and were injected intraperitoneally with normal saline 50 mg / kg 100 mg / kg and 200 mg / kg intraperitoneal injection of intraperitoneal bilirubin. After 24 hours of experimental treatment, blood samples were taken from the brain and heart of rats, bilirubin level and S-100 尾 concentration in serum and brain tissue were measured, pathological changes of hippocampal neurons were observed by HE staining, apoptosis of neurons was measured by Tunel method. The apoptotic rate was calculated and the experimental results of each index were recorded in time. The relevant data were calculated and statistically analyzed, and the conclusion was reached. To further evaluate the reliability and stability of neonatal rat model. Results 1) observe the general situation of experimental animals: before the experiment, the rats in each group were generally in good condition, the skin color was ruddy, elastic, sensitive to external stimuli, normal activity. After 24 hours of intervention, the skin color of the normal saline group was ruddy, the skin of the 50 mg / kg group was less ruddy and the elasticity was poor, and the skin color of the newborn rats in the 100mg / kg group and the 150 mg/kg group were yellowish. The skin of the rats in the 200 mg/kg group was yellowish green and had extremely poor elasticity. No response to external stimuli or even coma with HE staining: the hippocampal neurons in the normal saline group were arranged neatly, the structure was intact, and slight swelling was observed between the neurons in the 50 mg / kg group of normal morphology, and the neuronal structure was disordered in the 100 mg / kg group and the 150mg/kg group. The cells were swollen and deformed, some of the nuclear chromatin was dense and concentrated, and the cytoplasmic cavitation of 200 mg / kg group was extremely swollen and deformed, and most of the nuclear chromatin were dense and concentrated. Serum and brain bilirubin level and S-100 尾 concentration: compared with normal saline group, The serum and brain bilirubin levels and S-100 尾 concentrations were significantly different in each group (P 0.05), and the S-100 尾 levels in serum and brain tissues were not significantly different from those in 50mg/kg group. The apoptotic index of each group was significantly different from that of 150mg/kg group (P 0.05), but there was no significant difference in apoptotic index between 200 mg / kg group and 150mg/kg group (P 0.05). Conclusion the hyperbilirubinemia animal model can be established by intraperitoneal injection of 50 mg/kg intraperitoneal bilirubin in 1 / 7 day SD rats with 1: 1% mg/kg and stable bilirubin encephalopathy model with intraperitoneal injection of 100mg/kg and 150 mg/kg intraperitoneal bilirubin) when intraperitoneal injection of bilirubin in 150 mg/kg crystal can be used to establish a stable model of bilirubin encephalopathy. The glial cells were decomposed obviously, so the injury index model of glial cells could be established by injecting 150 mg/kg crystal bilirubin into SD rats on day 7.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R722.1;R-332

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