[Interleukin-1beta upregulates matrix metalloproteinase-2 expression and activity in cardiac fibroblasts via nitric oxide synthase pathway].
OBJECTIVE
To investigate the effect of interleukin-1beta (IL-1beta) on expression and activity of matrix metalloproteinase-2 (MMP-2) of cultured human cardiac fibroblasts and related signaling pathway.


METHODS
Primary human cardiac fibroblasts 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-1beta at various concentrations for 24 h, culture supernatant and cell protein were obtained. MMP-2 mRNA was determined by RT-PCR. The activity of MMP-2 was analyzed by zymography and the expression of inducible nitric oxide synthase (iNOS) protein level was detected by Western blot analysis. Assessment of NO production in the culture supernatant was performed using the Griess method.


RESULTS
IL-1beta (4 ng/ml) significantly increased MMP-2 activity of cultured fibroblasts in a time-dependent manner. MMP-2 mRNA expression was significantly upregulated by IL-1beta (4 ng/ml and 10 ng/ml, all P<0.01). Moreover, IL-1beta 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-1beta.


CONCLUSIONS
IL-1beta increased MMP-2 activity and transcription of human cardiac fibroblasts via iNOS-NO pathway.
Guo
Xiao-gang
XG
Department of Cardiology, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China. guoxiaogang11@yahoo.com.cn
Chen
Jun-zhu
JZ
Zhu
Jian-hua
JH
Zhang
Fu-rong
FR
Qiu
Yuan-gang
YG
Zhao
Li-li
LL
Shang
Yun-peng
YP
D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism
D053583:Interleukin-1beta; D012333:RNA, Messenger; D009569:Nitric Oxide; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2