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

冯洁; 李梦文; 韦倩; 李胜君; 华子瑜

doi:10.16016/j.1000-5404.201702007

您当前的位置：

首页 >

文章列表页 >

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

基础医学 | 更新时间：2026-01-09

- 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
- Role of caspase-1 activation in bilirubin neurotoxicity in vivo and in vitro
- 第三军医大学学报 2017年39卷第18期页码：1796-1802
- 作者机构：
  
  400014 重庆，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
- 作者简介：
  
  [ 华子瑜，E-mail: h_ziyu@163.com HUA Ziyu, E-mail: h_ziyu@163.com
- 基金信息：
  
  国家自然科学基金青年科学基金(81200459);重庆市基础科学与前沿技术研究一般项目(CSTC2013jcyjA0020)
- DOI：10.16016/j.1000-5404.201702007
  中图分类号： R345; R349.29; R722.17
- 收稿：2017-02-03，
  
  修回：2017-03-23，
  
  纸质出版：2017-09-30
- 稿件说明：
移动端阅览
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.

Jie FENG, Mengwen LI, Qian WEI, et al. Role of caspase-1 activation in bilirubin neurotoxicity in vivo and in vitro[J]. Journal of Third Military Medical University, 2017, 39(18): 1796-1802.
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. DOI： 10.16016/j.1000-5404.201702007.

Jie FENG, Mengwen LI, Qian WEI, et al. Role of caspase-1 activation in bilirubin neurotoxicity in vivo and in vitro[J]. Journal of Third Military Medical University, 2017, 39(18): 1796-1802. DOI： 10.16016/j.1000-5404.201702007.

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；VX-765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(

0.05)，IL-1β水平增高(

0.01)，脑组织切片皮层区星型胶质细胞活化(

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(

0.01)，生活能力改善(

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(

0.05)；与胆红素组相比，VX-765干预可抑制caspase-1活化(

0.05)，提高细胞存活率(

0.05)，减少培养液上清IL-1β的释放(

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 (

=42)

control group (

=30) and VX-765 treatment group (

=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 (

0.05) and content of IL-1β (

0.01) were significantly higher in the kernicterus model group

and the activation of astrocytes in the cortex of brain tissue was observed (

0.05)

when compared with the control group. Meanwhile

VX-

765 intervention resulted in decreased abnormal neural behaviors (

0.01) and improved life activities (

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 (

0.05)

but VX-765 intervention suppressed the activation of caspase-1 (

0.05)

increased the cell survival (

0.05)

and reduced the release of IL-1β in the supernatant (

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.

关键词

Keywords

references

BHUTANI V K, JOHNSON L. Synopsis report from the pilot USA kernicterus registry[J]. J Perinatol, 2009, 29(Suppl 1): S4-S7. DOI: 10.1038/jp.2008.210.

SGRO M, CAMPBELL D, SHAH V. Incidence and causes of severe neonatal hyperbilirubinemia in Canada[J]. CMAJ, 2006, 175(6): 587-590. DOI:10.1503/cmaj.060328.

TIRIBELLI C, OSTROW J D. The molecular basis of bilirubin encephalopathy and toxicity: report of an EASL Single Topic Conference, Trieste, Italy, 1-2 October, 2004[J]. J Hepatol, 2005, 43(1): 156-166. DOI: 10.1016/j.jhep. 2005.04.003.

GICQUEL T, ROBERT S, VICTONI T, et al. The NLRP3 inflammasome: physiopathology and therapeutic application[J]. Presse Med, 2016, 45(4 Pt 1): 438-446. DOI:10.1016/j.lpm.2016.01.006.

SINGHAL G, JAEHNE E J, CORRIGAN F, et al. Inflammasomes in neuroinflammation and changes in brain function: a focused review[J]. Front Neurosci, 2014, 8: 315. DOI: 10.3389/fnins.2014.00315.

SUN Q, SCOTT M J. Caspase-1 as a multifunctional inflammatory mediator: noncytokine maturation roles[J]. J Leukoc Biol, 2016, 100(5): 961-967. DOI:10.1189/jlb.3MR0516-224R.

Sollberger G, Strittmatter G E, Garstkiewicz M, et al. Caspase-1: the inflammasome and beyond[J]. Innate Immun, 2014, 20(2): 115-125. DOI: 10.1177/1753425 913484374.

SONG S, HU Y, GU X, et al. A novel newborn rat kernicterus model created by injecting a bilirubin solution into the cisterna magna[J]. PLoS ONE, 2014, 9(5): e96171. DOI: 10.1371/journal.pone.0096171.

李胜君, 李梦文, 张燕, 等.反式激活蛋白-NEMO结合域抑制胆红素诱导的大鼠皮层星形胶质细胞NF-κB活化[J].第三军医大学学报, 2015, 37(21): 2131-2136. DOI: 10.16016/j.1000-5404.201503068.

LI S J, LI M W, ZHANG Y, et al. Tat-nbd exerts anti-inflammatory effect in rat cortical astrocytes by inhibiting bilirubin-induced nuclear factor-κb activation[J]. J Third Mil Med Univ, 2015, 37(21): 2131-2136. DOI: 10.16016/j. 1000-5404.201503068.

FERNANDES A, BRITES D. Contribution of inflammatory processes to nerve cell toxicity by bilirubin and efficacy of potential therapeutic agents[J]. Curr Pharm Des, 2009, 15(25): 2915-2926. DOI:10.2174/138161209789058165.

BERGER I, PELEG O, OFEK-SHLOMAI N. Inflammation and early brain injury in term and preterm infants[J]. Isr Med Assoc J, 2012, 14(5): 318-323.

FERNANDES A, FALCÄO A S, SILVA R F, et al. Inflammatory signalling pathways involved in astroglial activation by unconjugated bilirubin[J]. J Neurochem, 2006, 96(6): 1667-1679. DOI: 10.1111/j.1471-4159.2006.03680.x.

BRITES D. The evolving landscape of neurotoxicity by unconjugated bilirubin: role of glial cells and inflammation[J]. Front Pharmacol, 2012, 3: 88. DOI: 10.3389/fphar.2012.00088.

SOFRONIEW M V, VINTERS H V. Astrocytes: biology and pathology[J]. Acta Neuropathol, 2010, 119(1): 7-35. DOI: 10.1007/s00401-009-0619-8.

ALLAMAN I, BÉLANGER M, MAGISTRETTI P J. Astrocyte-neuron metabolic relationships: for better and for worse[J]. Trends Neurosci, 2011, 34(2): 76-87. DOI: 10. 1016/j.tins.2010.12.001.

FERNANDES A, BARATEIRO A, FALCÃO A S, et al. Astrocyte reactivity to unconjugated bilirubin requires TNF-α and IL-1β receptor signaling pathways[J]. Glia, 2011, 59(1): 14-25. DOI:10.1002/glia.21072.

FERNANDES A, FALCãO A S, SILVA R F, et al. MAPKs are key players in mediating cytokine release and cell death induced by unconjugated bilirubin in cultured rat cortical astrocytes[J]. Eur J Neurosci, 2007, 25(4): 1058-1068. DOI: 10.1111/j.1460-9568.2007.05340.x.

LI M, SONG S, LI S, et al. The blockade of NF-κB activation by a specific inhibitory peptide has a strong neuroprotective role in a sprague-dawley rat kernicterus model[J]. J Biol Chem, 2015, 290(50): 30042-30052. DOI: 10.1074/jbc.M115.673525.

FANTUZZI G, DINARELLO C A. Interleukin-18 and interleukin-1 beta: two cytokine substrates for ICE (caspase-1)[J]. J Clin Immunol, 1999, 19(1): 1-11.

ALBONI S, CERVIA D, SUGAMA S, et al. Interleukin 18 in the CNS[J]. J Neuroinflammation, 2010, 7: 9. DOI: 10. 1186/1742-2094-7-9.

SEQUEIRA D, WATCHKO J F, DAOOD M J, et al. Unconjugated bilirubin efflux by bovine brain microvascular endothelial cells in vitro[J]. Pediatr Crit Care Med, 2007, 8(6): 570-575. DOI: 10.1097/01.PCC.0000288716.63685.1B.

BERGSBAKEN T, FINK S L, COOKSON B T. Pyroptosis: host cell death and inflammation[J]. Nat Rev Microbiol, 2009, 7(2): 99-109. DOI:10.1038/nrmicro2070.

STEVENSON D, MAISELS M J, WATCHKO J. Care of the Jaundiced Neonate[M]. New York: McGraw-Hill Professio nal, 2012: 1-285.

BAUERNFEIND F G, HORVATH G, STUTZ A, et al. Cutting edge: NF-kappaB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression[J]. J Immunol, 2009, 183(2): 787-791. DOI:10.4049/jimmunol.0901363.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

反式激活蛋白-NEMO结合域抑制胆红素诱导的大鼠皮层星形胶质细胞NF-κB活化

MAVS功能丧失性突变导致抗病毒免疫缺陷：揭示病毒感染触发儿童泛发性脓疱性银屑病的新机制

Z-藁本内酯通过抑制PI3K/AKT信号通路延缓视网膜色素变性：基于网络药理学与分子对接验证

PCBP1调控铁稳态抑制铁死亡对抗镉神经毒性的作用研究

ATF3通过抑制ATG5和ATG16L1表达调控巨噬细胞自噬与炎症反应