目的 研究新生大鼠高胆红素血症模型脑组织中NOD样受体蛋白3(NLRP3)小体活化与神经损害的相关性。方法 选择3日龄的SPF级新生SD大鼠36只,随机分为对照组、模型组和NLRP3小体抑制剂VX-765组(VX组)各12只,模型组和VX组通过腹腔注射100 μg/g胆红素的方式建立新生大鼠高胆红素血症模型,VX组同时给予NLRP3小体抑制剂VX-765腹腔注射。比较3组间血清及脑组织中胆红素含量、脑组织含水量及ATP含量、Nissl染色及TUNEL染色、血清NSE及S100B含量、NLRP3炎性小体表达的差异。结果 模型组大鼠脑组织中总胆红素含量、含水量、TUNEL阳性染色染色细胞率、NLRP3、ASC、cleaved-caspase-1、active-IL-1β、active-IL-18的表达水平及血清中总胆红素、NSE、S100含量明显高于对照组,脑组织中ATP含量及Nissl阳性染色细胞数目明显低于对照组(P<0.05);VX组大鼠脑组织中含水量、TUNEL阳性染色细胞率、cleaved-caspase-1、active-IL-1β、active-IL-18的表达水平及血清中NSE、S100含量明显低于模型组(P<0.05),脑组织中ATP含量及Nissl阳性细胞数目显著高于对照组(P<0.05)。结论 新生大鼠高胆红素血症模型中NLRP3小体活化参与了神经损伤过程。
Abstract
Objective To study the correlation between the activation of NOD-like receptor protein 3 (NLRP3) inflammasome and neurological damage in neonatal rat hyperbilirubinemia model.Methods A total of 36 3-days-old SPF-grade neonatal SD rats were randomly divided into control group,model group and VX group,with 12 rats in each group. The hyperbilirubinemia model of neonatal rats was established by intraperitoneal injection of 100 μg/g bilirubin in model group and VX group. The VX group was also given intraperitoneal injection of NLRP3 inflammasome inhibitor VX-765. The differences on bilirubin content in brain and serum,water content and ATP content in brain,Nissl staining and TUNEL staining,NSE and S100B contents in serum and the expression of NLRP3 inflammasome were analyzed among the three groups. Results The total bilirubin content,water content,TUNEL positive staining cell rate,NLRP3,ASC,cleaved-caspase-1,active-IL-1β,active-IL-18 expression levels in brain and serum total bilirubin,NSE,S100B contents in the model group were significantly higher than those in the control group (P<0.05). However,ATP content and the number of Nissl positive staining cells in the brain tissue were significantly lower than those in control group (P<0.05). The water content,TUNEL positive staining cell rate,cleaved-caspase-1,active-IL-1β,active-IL-18 expression level in brain and serum NSE,S100B contents in VX group were significantly lower than those in model group(P<0.05). ATP content and the number of Nissl positive staining cells in the brain tissue were significantly higher than those in model group (P<0.05). Conclusion NLRP3 inflammasome is involved in the process of nerve injury in neonatal rats with hyperbilirubinemia.
关键词
新生儿高胆红素血症 /
NOD样受体蛋白3 /
炎性小体 /
神经损害
Key words
neonatal hyperbilirubinemia /
NOD-like receptor protein 3 /
inflammasome /
neurological damage
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参考文献
[1] Bahr TM,Christensen RD,Agarwal AM,et al. The Neonatal Acute Bilirubin Encephalopathy Registry(NABER):background,aims,andprotocol[J].Neonatology,2019,115(3):242-246.
[2] Olusanya BO,Kaplan M,Hansen TWR.Neonatal hyperbilirubinaemia:a global perspective[J].Lancet Child Adolesc Health,2018,2(8):610-620.
[3] Rawat V,Bortolussi G,Gazzin S,et al. Bilirubin-induced oxidative stress leads to DNA damage in the cerebellum of hyperbilirubinemic neonatal mice and activates DNA double-strand break repair pathways in human cells[J].Oxid Med Cell Longev,2018,26(2018):1801243.
[4] Vodret S,Bortolussi G,Jašprová J,et al. Inflammatory signature of cerebellar neurodegeneration during neonatal hyperbilirubinemia in Ugt1 -/- mouse model[J].J Neuroinflammation,2017,14(1):64.
[5] Watchko JF. Bilirubin-induced neurotoxicity in the preterm neonate[J].Clin Perinatol,2016,43(2):297-311.
[6] 韦倩,冯洁,何春梅,等. Caspase-1活化在胆红素诱导的大鼠海马神经元损伤中的作用[J]. 南方医科大学学报,2018,38(5):567-571.
[7] 王晓丽,郭明星,梁俊晖,等. 新生大鼠高胆红素血症及脑病模型的建立与评价[J]. 中国病理生理杂志,2014,30(8):1523-1531.
[8] Helal NF,Ghany EAGA,Abuelhamd WA,et al. Characteristics and outcome of newborn admitted with acute bilirubin encephalopathy to a tertiary neonatal intensive care unit[J].World J Pediatr,2019,15(1):42-48.
[9] LePichon JB,Riordan SM,Watchko J,et al. The neurological sequelae of neonatal hyperbilirubinemia:definitions,diagnosis and treatment of the kernicterus spectrum disorders (KSDs)[J].Curr Pediatr Rev,2017,13(3):199-209.
[10] 张莹,艾雪梅,梁庆红,等. 胆红素脑病听觉受损的危险因素及防治策略[J]. 临床荟萃,2015,30(7):837-840.
[11] Wang D,Tosevska A,Heiβ EH,et al. Bilirubin decreases macrophage cholesterol efflux and ATP-binding cassette transporter A1 protein expression[J].J Am Heart Assoc,2017,6(5):e005520.
[12] Dal Ben M,Bottin C,Zanconati F,et al. Evaluation of region selective bilirubin-induced brain damage as a basis for a pharmacological treatment[J].Sci Rep,2017,19(7):41032.
[13] Viktorinova A.Iron-mediated oxidative cell death is a potential contributor to neuronal dysfunction induced by neonatal hemolytic hyperbilirubinemia[J].Arch BiochemBiophys,2018,15(654):185-193.
[14] Zhang XH,Zhang BL,Guo SM,et al. Clinical significance of dynamic measurements of seric TNF-α,HMGBl,and NSE levels and aEEG monitoring in neonatal asphyxia[J].Eur Rev Med Pharmacol Sci,2017,21(19):4333-4339.
[15] Zaigham M,Lundberg F,Olofsson P.Protein S100B in umbilical cord blood as a potential biomarker of hypoxic-ischemic encephalopathy in asphyxiated newborns[J].Early Hum Dev,2017,112:48-53.
[16] Wen L,Zhang QS,Heng Y,et al. NLRP3 inflammasome activation in the thymus of MPTP-induced parkinsonian mouse model[J].Toxicol Lett,2018,15(288):1-8.
[17] Liu W,Chen Y,Meng J,et al. Ablation of caspase-1 protects against TBI-induced pyroptosis in vitro and in vivo[J].J Neuroinflammation,2018,15(1):48.
[18] Jing S,Chi L,He Z,et al. NLRP3 inflammasome contributes to neurovascular unit damage in stroke[J].J Drug Target,2019,2:1-22.
[19] Zhu W,Cao FS,Feng J,et al. NLRP3 inflammasome activation contributes to long-term behavioral alterations in mice injected with lipopolysaccharide[J].Neuroscience,2017,343:77-84.
基金
上海市第六人民医院东院院级科学研究基金计划(DY2018002)
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