氧糖剥夺损伤诱导星形胶质细胞分泌IL-10的机制初探

吕泽宇; 何慕兰; 石霞; 张赟; 杨亭; 李廷玉; 陈洁

doi:10.13406/j.cnki.cyxb.001396

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氧糖剥夺损伤诱导星形胶质细胞分泌IL-10的机制初探

神经病学 | 更新时间：2024-01-17

- 氧糖剥夺损伤诱导星形胶质细胞分泌IL-10的机制初探
- Oxygen glucose deprivation injury induces IL-10 secretion from astrocytes and its mechanism
- 重庆医科大学学报 2017年42卷第10期页码：1227-1233
- 作者机构：
  
  重庆医科大学附属儿童医院儿童营养研究中心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
- 作者简介：
- 基金信息：
  
  国家自然科学基金资助项目(编号:81300522、81271385)
- DOI：10.13406/j.cnki.cyxb.001396
  中图分类号： R742
- 纸质出版日期：2017，
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吕泽宇, 何慕兰, 石霞, 等. 氧糖剥夺损伤诱导星形胶质细胞分泌IL-10的机制初探[J]. 重庆医科大学学报, 2017,42(10):1227-1233.

Lü Zeyu, He Mulan, Shi Xia, et al. Oxygen glucose deprivation injury induces IL-10 secretion from astrocytes and its mechanism[J]. Journal of Chongqing Medical University, 2017,42(10):1227-1233.
吕泽宇, 何慕兰, 石霞, 等. 氧糖剥夺损伤诱导星形胶质细胞分泌IL-10的机制初探[J]. 重庆医科大学学报, 2017,42(10):1227-1233. DOI： 10.13406/j.cnki.cyxb.001396.

Lü Zeyu, He Mulan, Shi Xia, et al. Oxygen glucose deprivation injury induces IL-10 secretion from astrocytes and its mechanism[J]. Journal of Chongqing Medical University, 2017,42(10):1227-1233. DOI： 10.13406/j.cnki.cyxb.001396.

摘要

目的:探讨氧糖剥夺(oxygen glucose deprivation

OGD)损伤诱导星形胶质细胞分泌白细胞介素-10(interleukin-10

IL-10)的可能机制。方法:培养并鉴定原代星形胶质细胞

体外建立OGD损伤模型

分别利用si TLR2(Toll-like receptor-2

TLR2)腺病毒、NF-κB(NF-kappa B)抑制剂PDTC干预

q RT-PCR和ELISA分别检测损伤细胞IL-10的表达水平

Western blot检测损伤细胞TLR2、磷酸化NF-κB p65(p-NF-κB p65)蛋白表达水平。免疫荧光检测OGD损伤细胞中NF-κB p65的亚细胞定位。结果:免疫荧光观察发现分离培养的原代星形胶质细胞纯度可达90%以上。经体外OGD损伤处理后

OGD损伤组TLR2、pNF-κB p65的蛋白表达水平[(1.16±0.10)

(1.01±0.08)]与IL-10的m RNA及蛋白表达水平[(0.00±0.00)

(513.01±34.47)]较正常对照组[(0.49±0.08)

(0.23±0.09)

(0.00±0.00)

(381.89±22.26)]均明显升高(P=0.001

P=0.000

P=0.026

P=0.005)

且p65蛋白在细胞核内表达增加。si TLR2腺病毒感染组OGD损伤的星形胶质细胞中TLR2、p-NF-κB p65的蛋白表达水平[(0.36±0.13)

(0.38±0.10)]与IL-10的m RNA及蛋白表达水平[(0.00±0.00)

(567.05±50.57)]较对照组[(1.33±0.42)

(0.92±0.28)

(0.00±0.00)

(703.60±12.97)]明显降低(P=0.019

P=0.034

P=0.004

P=0.011)。与OGD处理组[(1.24±0.23)

(0.00±0.00)

(769.23±23.37)]相比

PDTC干预可有效降低p-NF-κB p65蛋白(0.26±0.07)与IL-10的m RNA及蛋白表达水平[(0.00±0.00)

(598.92±42.73)](P=0.002

P=0.000

P=0.046)

但并未影响TLR2的表达水平变化[(1.11±0.24)

(1.09±0.94)](P=0.949)。结论:OGD损伤可激活TLR2/NF-κB信号通路诱导星形胶质细胞分泌IL-10。

Abstract

Objective:To investigate the potential mechanism involved in interleukin（IL-10）secretion from astrocytes when being treated with oxygen glucose deprivation（OGD）. Methods:Primary astrocytes isolated from neonatal Sprague Dawley rats were cultivated and identified as usual methods. After being suffered from OGD injury

the cultured astrocytes were treated with either vehicle

si TLR2（Toll-like receptor-2

TLR2）or PDTC（a NF-κB inhibitor）

respectively. First

real-time PCR and ELISA techniques were employed to examine the levels of IL-10 transcription and expression in the OGD-injured astrocytes. Next

the protein levels of TLR2 and pNF-κB p65 among these OGD-injured astrocytes were detected using Western blot. In addition

the subcellular distribution of NF-κB p65 in OGD-injured astrocytes was tested with immunofluorescence. Results:Immunofluorescence revealed that the isolated primary cultured astrocytes reached a high purity more than ninety percent. Compared with the control group[（0.49±0.08）

（0.23±0.09）

（0.00 ±0.00）

（381.89 ±22.26）]

OGD treatment not only upregulated the TLR2

p-NF-κB p65 and IL-10 expression levels[（1.16 ±0.10）

（1.01 ±0.08）

（0.00 ±0.00）

（513.01 ±34.47）] among these astrocytes（P =0.001

P =0.000

P =0.026

P =0.005）

but also increased the protein expression level of NF-κB p65 in the nucleus. However

TLR2 si RNA effectively reduced the TLR2 expression level and prevented OGD-induced p-NF-κB p65 and IL-10 upregulation [（0.36 ±0.13）

（0.38 ±0.10）

（0.00±0.00）

（567.05±50.57）] when compared with those in si TLR2-untreatment group [（1.33±0.42）

（0.92±0.28）

（0.00±0.00）

（703.60±12.97）]. Additionally

PDTC also dramatically decreased p-NF-κB p65 and IL-10 expression levels[（0.26±0.07）

（0.00±0.00）

（598.92±42.73）] without affecting TLR2 expression in OGD-injured astrocytes（1.09±0.94）comparing with those in OGD group[（1.24±0.23）

（0.00±0.00）

（769.23±23.37）

（1.11±0.24）]（P=0.002

P=0.000

P=0.046

P=0.949）. Conclusion:TLR2/NF-κB signaling pathway may mediate OGD-induced IL-10 secretion from astrocytes.

关键词

星形胶质细胞白细胞介素-10氧糖剥夺Toll样受体2核因子-κB

Keywords

primary astrocytesinterleukin-10oxygen glucose deprivationToll-like receptor-2nuclear factor κB

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