经向小脑延髓池内注射胆红素溶液建立的新生大鼠胆红素脑病模型
A novel newborn rat kernicterus model created by injecting a bilirubin solution into the cisterna magna.
机构信息
Department of Neonatology, Children's Hospital of Chongqing Medical University, Chongqing, China; Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Medical University, Chongqing, China; Chongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory Disorders, Chongqing Medical University, Chongqing, China; Key Laboratory of Pediatrics in Chongqing, Chongqing Medical University, Chongqing, China; Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital, Chongqing Medical University, Chongqing, China.
Department of Neonatology, Children's Hospital of Chongqing Medical University, Chongqing, China; Department of Pediatrics, Southwest Hospital, Third Military Medical University, Chongqing, China.
出版信息
PLoS One. 2014 May 5;9(5):e96171. doi: 10.1371/journal.pone.0096171. eCollection 2014.
BACKGROUND
Kernicterus still occurs around the world; however, the mechanism of bilirubin neurotoxicity remains unclear, and effective treatment strategies are lacking. To solve these problems, several kernicterus (or acute bilirubin encephalopathy) animal models have been established, but these models are difficult and expensive. Therefore, the present study was performed to establish a novel kernicterus model that is simple and affordable by injecting unconjugated bilirubin solution into the cisterna magna (CM) of ordinary newborn Sprague-Dawley (SD) rats.
METHODS
On postnatal day 5, SD rat pups were randomly divided into bilirubin and control groups. Then, either bilirubin solution or ddH2O (pH = 8.5) was injected into the CM at 10 µg/g (bodyweight). For model characterization, neurobehavioral outcomes were observed, mortality was calculated, and bodyweight was recorded after bilirubin injection and weaning. Apoptosis in the hippocampus was detected by H&E staining, TUNEL, flow cytometry and Western blotting. When the rats were 28 days old, learning and memory ability were evaluated using the Morris water maze test.
RESULTS
The bilirubin-treated rats showed apparently abnormal neurological manifestations, such as clenched fists, opisthotonos and torsion spasms. Bodyweight gain in the bilirubin-treated rats was significantly lower than that in the controls (P<0.001). The early and late mortality of the bilirubin-treated rats were both dramatically higher than those of the controls (P = 0.004 and 0.017, respectively). Apoptosis and necrosis in the hippocampal nerve cells in the bilirubin-treated rats were observed. The bilirubin-treated rats performed worse than the controls on the Morris water maze test.
CONCLUSION
By injecting bilirubin into the CM, we successfully created a new kernicterus model using ordinary SD rats; the model mimics both the acute clinical manifestations and the chronic sequelae. In particular, CM injection is easy to perform; thus, more stable models for follow-up study are available.
背景
核黄疸仍在全球范围内发生；然而，胆红素神经毒性的机制仍不清楚，也缺乏有效的治疗策略。为了解决这些问题，已经建立了几种核黄疸（或急性胆红素脑病）动物模型，但这些模型既困难又昂贵。因此，本研究通过向普通新生 Sprague-Dawley（SD）大鼠的枕骨大孔（CM）内注射未结合胆红素溶液，建立了一种简单且经济实惠的新型核黄疸模型。
方法
在出生后第 5 天，SD 大鼠幼仔被随机分为胆红素组和对照组。然后，向 CM 内分别注射胆红素溶液或 ddH2O（pH=8.5），剂量为 10μg/g（体重）。为了对模型进行特征描述，观察神经行为学结果，计算死亡率，并在胆红素注射和断奶后记录体重。通过 H&E 染色、TUNEL、流式细胞术和 Western blot 检测海马细胞凋亡。当大鼠 28 天时，使用 Morris 水迷宫测试评估学习和记忆能力。
结果
胆红素处理的大鼠表现出明显异常的神经表现，如紧握拳头、角弓反张和扭转痉挛。胆红素处理的大鼠体重增长明显低于对照组（P<0.001）。胆红素处理的大鼠的早期和晚期死亡率均明显高于对照组（P=0.004 和 0.017）。胆红素处理的大鼠海马神经细胞出现凋亡和坏死。胆红素处理的大鼠在 Morris 水迷宫测试中的表现明显差于对照组。
结论
通过向 CM 内注射胆红素，我们成功地使用普通 SD 大鼠建立了一种新的核黄疸模型；该模型模拟了急性临床表现和慢性后遗症。特别是，CM 注射易于操作；因此，为后续研究提供了更稳定的模型。
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