Interleukin-1β upregulates matrix metalloproteinase-2 expression and activity in cardiac fibroblastsvia nitric oxide synthase pathway
10.3760/cma.j.issn.0253-3758.2009.01.014
- VernacularTitle:诱导型一氧化氮合酶参与白细胞介素-1β对人心脏成纤维细胞基质金属蛋白酶-2作用的实验研究
- Author:
Xiao-Gang GUO
1
;
Jun-Zhu CHEN
;
Jian-Hoa ZHU
;
Fu-Rong ZHANG
;
Yuan-Gang QIU
;
Li-Li ZHAO
;
Yun-Peng SHANG
Author Information
1. 浙江大学医学院附属第一医院
- Keywords:
Heart;
Fibroblasts;
Intedeukin-1beta;
Nitric oxide synthase;
Matrix metalloproteinases
- From:
Chinese Journal of Cardiology
2009;37(1):63-68
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of interleukin-1β(IL-1β)on expression and activity of matrix metallopmteinage-2(MMP-2)of cultured human cardiac fibroblasts and related signaling pathway.Methods Primary human cardiac fibmblasts seeded in 6-well tissue culture plates and cultured to 80%to 90%confluence were harvested at passage 3 to 6 and exposed to IL-1β at vaious concentrations for 24 h,culture supernatant and cell protein were obtained.MMP-2 mRNA was determined by RT-PCR.The activity of MMP-2 wag analyzed by zymography and the expression of inducible nitric oxide synthese(iNOS)Drotein level was detected by Western blot analysis.Assessment of NO production in the culture supernatant wag performed using the Griess method.Results IL-1β(4 ng/ml)significandy increased MMP-2 activity of cultured fibmblagts in a time-dependent manner.MMP-2 mRNA expression wag significantly upregulated by IL-1β(4 ng/ml and 10 ng/ml,all P＜0.01).Moreover,IL-1β also significantly increased NO production in supernatant(P＜0.01)and these effects could be significantly blocked by cotreatment with L-NMMA(10-3 mol/L,all P＜0.01).Western blot analysis showed that iNOS could not be detected in unstimulated human cardiac fibroblasts but could be detected in cardiac fibroblasts exposed to IL-1β.Conclusion IL-1β increased MMP-2 activity and transcription of human cardiac fibroblasts via iNOS-NO
