[1]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.
 Li Shengjun,Li Mengwen,Zhang Yan,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.
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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(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
37卷
期数:
2015年第21期
页码:
2131-2136
栏目:
论著
出版日期:
2015-11-15

文章信息/Info

Title:
TAT-NBD exerts anti-inflammatory effect in rat cortical astrocytes by inhibiting bilirubin-induced nuclear factor-κB activation
作者:
李胜君李梦文张燕冯洁华子瑜
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,儿科学重庆市重点实验室
Author(s):
Li Shengjun Li Mengwen Zhang Yan Feng Jie Hua Ziyu

Department of Neonatology, Key Laboratory of Developmental Diseases in Children of Ministry of Education, Chongqing Key Laboratory of Pediatrics, Children’s Hospital of Chongqing Medical University, Chongqing, 400014, China

关键词:
胆红素星形胶质细胞核因子-&kappaB炎症TAT-NBD
Keywords:
bilirubin cortical astrocytes nuclear factor-kappa B inflammation trans-activator of transcription-NEMO binding domain
分类号:
R392.32; R742.89; R977.6
文献标志码:
A
摘要:

目的      明确反式激活蛋白-NEMO结合域(TAT-NBD)对胆红素诱导的大鼠皮层星形胶质细胞核因子-κB(NF-κB)活化及炎症反应的作用。      方法       分离并鉴定新生SD大鼠大脑皮质星形胶质细胞,随机分为对照组、胆红素组、TAT-NBD干预组。免疫荧光观察各组细胞形态,改良MTT检测细胞相对存活率。Western blot检测NF-κB p65表达,EMSA检测NF-κB的入核、活化。ELISA法检测培养基上清炎症因子IL-1β、TNF-α、IL-6水平。      结果      胆红素组NF-κB p65蛋白灰度比值明显高于对照组(P<0.01),2、12 h达高峰(P<0.05);TAT-NBD干预组NF-κB p65蛋白表达明显低于胆红素组(P<0.01)。IL-1β、TNF-α、IL-6的释放与胆红素诱导的NF-κB p65表达有时间相关性,于12 h达高峰,且TAT-NBD干预2、12 h的IL-1β、TNF-α、IL-6 浓度均低于胆红素组(P<0.01)。星形胶质细胞的存活率与胆红素作用时间呈负相关,TAT-NBD干预2、12 h的相对存活率均高于胆红素组(P<0.05)。      结论       胆红素可诱导原代培养的大鼠星形胶质细胞NF-κB激活、过表达,TAT-NBD抑制NF-κB的活化高峰、减少炎症因子释放产生抗炎作用,可用于预防胆红素脑损伤。

Abstract:

Objective       To determine the effect of trans-activator of transcription-NEMO binding domain (TAT-NBD) on bilirubin-induced activation of nuclear factor-κB (NF-κB) and neuroinflammation in rat cortical astrocytes.       Methods       Astrocytes were freshly separated from the cortex of newborn SD rats. After identification and purification, the astrocytes at passage 2 were randomly divided into control group, bilirubin group and TAT-NBD intervention group. Cellular morphology were observed by immunofluorescence assay, the survival rate were assessed with a modified MTT assay, the expression and activity of NF-κB p65 protein was detected by Western blotting and electrophoretic mobility shift assay (EMSA), and the contents of IL-1β, TNF-α and IL-6 in the supernatants were measured with ELISA.       Results       Compared with the astrocytes in the control group, the expression of NF-κB p65 protein was significantly increased in bilirurin-treated cells (P<0.01), reaching the peak at 2 and 12 h (P<0.05). Consistent with the expression of NF-κB, the contents of IL-1β, TNF-α and IL-6 were in a time-dependent manner. TAT-NBD intervention dramatically inhibited bilirubin-induced NF-κB p65 activation as well as the release of IL-1β, TNF-α and IL-6 (P<0.01). The viability of astrocyte cells was decreased after bilirubin exposure, whereas the survival rate of TAT-NBD intervention group was significantly higher than that of bilirubin group but lower than that of the control group (P<0.01).       Conclusion      In in vitro model, bilirubin-induced cortical astrocytes inflammation is obviously related to the over-expression and activation of NF-κB. TAT-NBD does show strong anti-inflammatory effects by inhibiting the activation of NF-κB and release of inflammatory cytokines, which might be useful in prophylaxis of bilirubin-induced brain injury.

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更新日期/Last Update: 2015-11-06