MAPK和NF-κB 上 调 脂 多 糖 诱 导 小 鼠 肺 微 血 管 内 皮 细 胞 Toll-like 受体4 的 表 达

基础医学与临床 ›› 2006, Vol. 26 ›› Issue (12): 1328-1331.

MAPK和NF-κB 上调脂多糖诱导小鼠肺微血管内皮细胞 Toll-like 受体4 的表达

余跃杜烨玮孙仁宇王士雯

中国医学科学院基础医学研究所解放军老年心血管研究所医科院基础医学研究所

收稿日期:2006-03-15 修回日期:2006-06-21 出版日期:2006-12-25 发布日期:2006-12-25
通讯作者: 余跃

MAPK and NF-κB up-regulated the expression of Toll-like receptor 4 induced by LPS in pulmonary micro-vascular endothelial cell in mouse

Received:2006-03-15 Revised:2006-06-21 Online:2006-12-25 Published:2006-12-25

摘要/Abstract

摘要： 目的观察 MAPK和 NF-κB 在经脂多糖 ( LPS ) 诱导的小鼠肺微血管内皮细胞 ( PWVEC ) Toll-like 受体4 ( T L R4 ) 表达增强中的作用。方法分离培养 PWVEC。不同浓度的 LPS 与 PWVEC 共培养 2 h , 或与 LPS ( 100 μg / L) 作用不同时间。用 ELISA 法检测培养上清液中TNF-α含量。分别用 ERK抑制剂PD98059 和 p38 MAPK抑制剂SB203580 或NF-κB抑制剂PDTC 进行干预。用反转录聚合酶链反应（RT-PCR）检测 TLR4 和TNF-αmRNA。用Western-blot检测 ERK / p38 MAPK 和核蛋白NF-κB 的表达。结果在 LPS 浓度为50~500μg / L 时 , 作用于PWVEC 2 h , 或 LPS 100 μg / L 作用不同时间时 , 培养上清液中T N F-α 含量呈剂量和时间依赖性显著增加 ( P< 0.01)。 PWVEC 表达 TNF-α及TLR4 mRNA 增加 , 6 h 时达高峰。分别用上述抑制剂均可缓解上述变化 , 而同时联合使用 PD98059 和SB203580 则进一步增强该抑制作用。上述抑制剂分别缓解 LPS 诱导的 PWVEC 表达 ERK / p38 MAPK 和NF-κB 蛋白增强。结论 MAPK 和NF-κB 上调脂多糖诱导小鼠PWVEC TLR4 的表达。

Abstract: Objective To explore the role of MAPK and NF-κB in increase Toll-like receptor 4 ( TLR4 ) expression induced by lipopolysaccharide (LPS ) in pulmonary micro-vascular endothelial cell (PMVEC) in mouse. Methods Isolation and culture mouse PMVEC. PMVEC were exposed to the different concentration LPS at 2h, and 100 μg/L LPS in different time. The concentration of TNF-αin culture supernatant was detected using ELISA. Using PD98059, ERK inhibitor, SB203580, inhibitor for p38 MAPK and PDTC inhibitor for NF-κB pretreated PMVEC. RT-PCR was used to detect the levels of TLR4 and TNF-αmRNA. Western-blot was used to detect the protein expression of nuclear factor-kappa B ( NF-κB ) p65 subunit and ERK / p38 MAPK in nuclear extract after LPS exposure for 6 h respectively. Results At doses of 50~500 ng /mL LPS for 2 h or 100μg / L LPS exposure in different time, the concentration of TNF-αin culture supernatant of PMVEC was increase with both time and dosage dependent ( P <0.01). The levels of TLR4 and TNF-αmRNA in membrane of PMVEC were obviously increase, with the peak at 6h. Above inhibiter decrease the TLR4 and TNF-αmRNA expression separately, and above inhibition effect was increased after simultaneous pretreatment with PD98059 and SB203580. Protein expression of ERK/p38 MAPK and NF-κB in PMVEC induced by LPS were down-regulation by pretreatment with PD98059, SB203580 and PDTC respectively. Conclusion Up-regulation expression of TLR4 in PWVEC of mouse by MAPK and NF-κB pathway induced by LPS.

余跃杜烨玮孙仁宇王士雯. MAPK和NF-κB 上调脂多糖诱导小鼠肺微血管内皮细胞 Toll-like 受体4 的表达[J]. 基础医学与临床, 2006, 26(12): 1328-1331.

. MAPK and NF-κB up-regulated the expression of Toll-like receptor 4 induced by LPS in pulmonary micro-vascular endothelial cell in mouse[J]. Basic & Clinical Medicine, 2006, 26(12): 1328-1331.