[1]冯洁,李梦文,韦倩,等.半胱氨酸天冬氨酸蛋白酶-1活化在胆红素神经毒性中的作用研究[J].第三军医大学学报,2017,39(18):1796-1802.
 FENG Jie,LI Mengwen,WEI Qian,et al.Role of caspase-1 activation in bilirubin neurotoxicity in vivo and in vitro[J].J Third Mil Med Univ,2017,39(18):1796-1802.
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半胱氨酸天冬氨酸蛋白酶-1活化在胆红素神经毒性中的作用研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第18期
页码:
1796-1802
栏目:
基础医学
出版日期:
2017-09-30

文章信息/Info

Title:
Role of caspase-1 activation in bilirubin neurotoxicity in vivo and in vitro
作者:
冯洁李梦文韦倩李胜君华子瑜
重庆医科大学附属儿童医院新生儿科,儿童发育疾病研究教育部重点实验室,儿童发育重大疾病国家国际科技合作基地,认知发育与学习记忆障碍转化医学重庆市重点实验室
Author(s):
FENG Jie LI Mengwen WEI Qian LI Shengjun HUA Ziyu

Department of Neonatology, Key Laboratory of Developmental Diseases in Children of Ministry of Education, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory Disorders, Children’s Hospital of Chongqing Medical University, Chongqing, 400014, China

关键词:
胆红素神经毒性星型胶质细胞半胱氨酸天冬氨酸蛋白酶-1炎症
Keywords:
bilirubin neurotoxicity astrocytes caspase-1 inflammation
分类号:
R345;R349.29;R722.17
文献标志码:
A
摘要:

目的     明确半胱氨酸天冬氨酸蛋白酶-1(caspase-1)活化是否参与胆红素神经毒性的发生机制;VX-765 抑制caspase1活化是否能抑制胆红素神经毒性,发挥神经保护作用。方法     建立胆红素脑病动物模型,Western blot 检测脑组织caspase-1蛋白的表达,ELISA 法检测炎症因子IL-1β的水平,免疫荧光染色检测脑组织中GFAP蛋白的表达;VX0765干预后动态观察各组新生鼠的神经系统临床表现,记录新生鼠体质量变化,评估生活能力。原代培养大鼠皮层星型胶质细胞分为胆红素组、VX-765组、对照组:改良MTT法检测细胞存活率,Western blot 检测细胞caspase-1蛋白的表达,ELISA 法检测培养液上清IL-1β的水平。结果     胆红素脑病动物模型,建模后12 h,胆红素组较对照组,脑组织中活化型caspase-1表达增加(P<0.05),IL-1β水平增高(P<0.01),脑组织切片皮层区星型胶质细胞活化(P<0.05)。与胆红素组相比,VX-765组新生鼠建模后异常神经系统表现减少(P<0.01),生活能力改善(P<0.05)。原代培养大鼠皮层星型胶质细胞,胆红素干预6h后,活化型caspase-1表达显著高于对照组(P<0.05);与胆红素组相比,VX-765干预可抑制caspase-1活化(P<0.05),提高细胞存活率(P<0.05),减少培养液上清IL-1β的释放(P<0.01)。结论      胆红素可诱导活化型caspase-1表达增加;VX-765抑制caspase-1活化可减轻胆红素神经毒性,提高原代培养星型胶质细胞存活率,减少炎症因子释放。

Abstract:

Objective     To investigate whether the activity of caspase-1 is involved in the pathogenesis of bilirubin neurotoxicity, and whether VX-765 (caspase1 specific inhibitor) exerts neuroprotective effect in the process. Methods      Neonatal SD rats were randomly divided into Kernicterus group (n=42), control group (n=30) and VX-765 treatment group (n=12, 50 mg/g, via intra peritoneal injection). Kernicterus model was established by injecting 1 μL/g bilirubin into posterior cistern in the rats. The expression of caspase-1 was assessed by Western blotting, the content of IL-1β in the brain tissues was measured-by ELISA, and the expression of GFAP protein was detected by immunofluorescence assay. Further, VX-765 was administered intraperitoneally at 24 h and 1 h before model establishment, typical neurological manifestations and body weight were dynamically recorded to evaluate their life activities. Primarily cultured rat cortical astrocytes were randomly divided into control group, bilirubin group and VX-765 intervention group. The survival rate of the astrocytes was assessed with modified MTT assay, expression of caspase-1 was detected by Western blotting, and content of IL-1β in the supernatant was measured by ELISA. Results     At 12 h after bilirubin injection, the expression of caspase-1 (P<0.05) and content of IL-1β (P<0.01) were significantly higher in the kernicterus model group, and the activation of astrocytes in the cortex of brain tissue was observed (P<0.05), when compared with the control group. Meanwhile, VX-765 intervention resulted in decreased abnormal neural behaviors (P<0.01) and improved life activities (P<0.05). In primarily cultured rat cortical astrocytes, bilirubin exposure for 6 h enhanced the expression of activated caspase-1 when compaired with the control group (P<0.05), but VX-765 intervention suppressed the activation of caspase-1 (P<0.05), increased the cell survival  (P<0.05), and reduced the release of IL-1β in the supernatant (P<0.01). Conclusion     Bilirubin induces the expression of activated caspase-1. While VX-765 alleviates the bilirubin-induced neurotoxicity by inhibiting the activation of caspase-1, and then improves the survival rate of primarily cultured rat cortical astrocytes and reduces the release of inflammatory factors.
 

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更新日期/Last Update: 2017-09-29