PrP106-126及Aβ1-42同步诱导BV-2小胶质细胞趋化及增殖活性的研究

doi:10.11843/j.issn.0366-6964.2014.10.017

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (10): 1699-1710.doi: 10.11843/j.issn.0366-6964.2014.10.017

PrP^106-126及Aβ_1-42同步诱导BV-2小胶质细胞趋化及增殖活性的研究

涂健^1,2，杨利峰¹，祁克宗²，周向梅¹，尹晓敏¹，赵德明^1*

（1.中国农业大学国家海绵状脑病实验室，北京 100193；2.安徽农业大学动物科学学院，合肥 230036）

收稿日期:2014-07-01 出版日期:2014-10-23 发布日期:2014-10-23
通讯作者: 赵德明，E-mail：zhaodm@cau.edu.cn
作者简介:涂健(1980-)，男，安徽安庆人，讲师，博士，主要从事动物疫病病理与生物防治研究，Tel：0551-65786305，E-mail：tujian1980@126.com
基金资助:
国家自然科学基金（30771622；31001048）

PrP^106-126 and Aβ_1-42 Peptides Induce Simultaneously BV-2 Microglia Chemotaxis and Proliferation

TU Jian^1,2，YANG Li-feng¹，QI Ke-zong²，ZHOU Xiang-mei¹，YIN Xiao-min¹，ZHAO De-ming^1*

（1.National Animal TSE Lab，College of Veterinary Medicine，China Agricultural University，Beijing 100193，China;2.College of Animal Science and Technology，Anhui Agricultural University，Hefei 230036，China）

Received:2014-07-01 Online:2014-10-23 Published:2014-10-23

摘要/Abstract

摘要：

通过检测传染性海绵状脑病（TSEs）及阿尔茨海默病（AD）的PrP和Aβ毒性蛋白同步诱导小胶质细胞趋化与增殖活性，旨在初步揭示TSEs及AD的致病机制及重新评估TSEs危害公共卫生安全的风险。以PrP^106-126和Aβ_1-42为研究对象，以BV-2小胶质细胞为实验细胞，构建25~100 μmol•L^－1的PrP_106-126和2.5~10 μmol•L^－1的Aβ_1-42以及两者不同浓度比例混合物侵染BV-2细胞的实验模型，比较多肽及其混合物诱导BV-2的趋化指数（CI）、增殖指数（PI）以及MCP-1、TGF-β1的表达水平，同步进行上述参数的相关性分析。结果表明：与PBS组及蛋白对照组相比，25~100 μmol•L^－1的PrP^106–126及2.5~10 μmol•L^－1Aβ_1-42均上调BV-2的CI、PI及表达MCP-1、TGF-β1水平（P<0.05）；两种多肽不同浓度混合物同步诱导BV-2的CI、PI值及MCP-1、TGF-β1表达水平均大于单一蛋白分别诱导相同参数值，但却小于两种蛋白分别诱导相同参数值之和；两者诱导BV-2 CI、PI及MCP-1表达水平均呈现多肽低浓度依赖性及处理时间依赖性，但当多肽达到高浓度或长诱导时间（PrP^106-126＞50 μmol•L^－1；Aβ_1-42＞5 μmol•L^－1；诱导时间＞12 h）时，这3种参数都进入平台期，但BV-2表达TGF-β1水平持续上升；PrP^106-126及Aβ_1-42对BV-2 RCI与 MCP-1水平呈正相关。结果提示，两种多肽均能上调BV-2的CI、PI及表达MCP-1、TGF-β1水平，并引起这些参数有规律的变化，两者在蛋白水平同步诱导BV-2上述参数有明显的拮抗作用。

Abstract:

Transmissible spongiform encephalopathies (TSEs) and Alzheimer’s disease (AD) belong to a growing family of neurodegenerative disorders that is characterized by the generation of toxic protein aggregates in affected brains (PrP^Sc and Aβ in TSEs and AD，respectively).At present，clinical and experimental evidence indicates the coexistence of PrP and Aβ amyloid deposits in affected brain tissues.Based on these pathological and mechanistic similarities，relationship between TSEs and AD should be further investigated.In this study，we examined the activation of BV-2 microglial chemotaxis and proliferation as well as BV-2 cell secretion of MCP-1 and TGF-β1 after treatment with aggregated forms of PrP^106-126 and Aβ_1-42 (separately and together) in vitro in an attempt to understand how protein aggregates can modulate microglial processes in TSEs and AD.We treated BV-2 microglia with PrP^106-126 or Aβ_1-42 peptides individually at thee different concentrations (25-100 μmol•L^－1 PrP^106-126 and 2.5-10 μmol•L^－1 Aβ_1-42) or with a mixture of PrP^106-126 and Aβ_1-42 peptides at specified concentrations for 6-24 h.BV-2 microglia chemotaxis，proliferation，and MCP-1 and TGF-β1 secretion were measured and compared between treatments.The results demonstrate that PrP^106-126 and Aβ_1-42 peptides induce increases in all four parameters from 6-12 h.However，the measured indices plateaued beyond 12 h in BV-2 cells treated >50 μmol•L^－1 PrP^106-126 or >5 μmol•L^－1 Aβ_1-42，with the exception of TGF-β1 secretion，which continued to increase gradually.Overall，the results of this study indicated that PrP^106-126 and Aβ_1-42 peptides induce increases in all four parameters from 6-12 h and these two peptides have obviously antagonistic effects in inducing microglial chemotaxis and proliferation simultaneously at the protein level.

中图分类号:

S852.659.7

涂健，杨利峰，祁克宗，周向梅，尹晓敏，赵德明. PrP^106-126及Aβ_1-42同步诱导BV-2小胶质细胞趋化及增殖活性的研究[J]. 畜牧兽医学报, 2014, 45(10): 1699-1710.

TU Jian，YANG Li-feng，QI Ke-zong，ZHOU Xiang-mei，YIN Xiao-min，ZHAO De-ming. PrP^106-126 and Aβ_1-42 Peptides Induce Simultaneously BV-2 Microglia Chemotaxis and Proliferation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(10): 1699-1710.

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PrP^106-126及Aβ_1-42同步诱导BV-2小胶质细胞趋化及增殖活性的研究

PrP^106-126 and Aβ_1-42 Peptides Induce Simultaneously BV-2 Microglia Chemotaxis and Proliferation

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PrP106-126及Aβ1-42同步诱导BV-2小胶质细胞趋化及增殖活性的研究

PrP106-126 and Aβ1-42 Peptides Induce Simultaneously BV-2 Microglia Chemotaxis and Proliferation

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PrP^106-126及Aβ_1-42同步诱导BV-2小胶质细胞趋化及增殖活性的研究

PrP^106-126 and Aβ_1-42 Peptides Induce Simultaneously BV-2 Microglia Chemotaxis and Proliferation