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Cardiovascular disease basic research
The effects of sPLA2-IIA in human umbilical vein endothelial cells
  1. Tan Donghan,
  2. He Mingyue,
  3. Chen Lianfeng,
  4. Fang Quan
  1. Peking Union Medical College Hospital, Dongcheng, Beijing, China

Abstract

Background Secretory phospholipase A2 group IIA (sPLA2-IIA) appears to be an important inflammatory mediator of cardiovascular disease and may play a pivotal role in the pathophysiology of AS. To understand whether the influence of sPLA2-IIA on proinflammantory effects in vascular endothelial cells may help comprehend the mechanism induced by the sPLA2-IIA involved in AS.

Objectives To investigate the effect of different concentrations of sPLA2-IIA on Endothelial Cells, and to investigate the signalling pathway.

Methods (1) HUVEC were cultured in three different concentrations groups (0.01, 0.1, 1 ug/ml) of sPLA2-IIA. Endothelial cell nitric oxide concentration in the supernatant was detected by kit. The mRNA expression levels of ET-1, eNOS, ICAM-1, VCAM-1 were determined by Real Time-PCR. The protein expression level were measured by Western Blot or by ELISA. (2) HUVEC cells were cultured with 1 ug/ml sPLA2-IIA and 10 nmol/l 4-BPB (Hydrolysis inhibitor) or 1 umol/l PD98059 (ERK1/2 inhibitor) or SP600125 (JNK inhibitor) or SB203580 (p38 inhibitor) or Bay11-7085 (NF-kB inhibitor) alone or combined. Parameters described in Method step 1 were measured.

Results (1) sPLA2-IIA increased the mRNA and protein expression of ICAM-1, VCAM-1 and ET-1 and decreased the level of NO in a concentration dependent way. 1 ug/ml of sPLA2-IIA dramatically decreased the mRNA expression of eNOS. The protein expression of eNOS was not affected by sPLA2-IIA. (2) 4-BPB abolished the over expression of mRNA and protein of ICAM-1, VCAM-1 and ET-1 induced by the sPLA2-IIA. 4-BPB also abolished the up- regulated mRNA of eNOS and reduced the down-regulation of NO induced by sPLA2-IIA. (3) PD98059 abolished the over expression of ICAM-1, VCAM-1 and ET-1 and reduced the down-regulation of NO induced by the sPLA2-IIA. (4) SP600125 abolished the over expression of ET-1 and reduced the down-regulation of NO induced by sPLA2-IIA. (5) SB203580 had little effect on the over expression of ICAM-1, VCAM-1 and ET-1 and down-regulation of NO induced by sPLA2-IIA. 6. Bay11-7085 abolished the over expression of ICAM-1, VCAM-1 and ET-1 induced by the sPLA2-IIA.

Conclusions (1) sPLA2-IIA induced the over expression of cytokines including ICAM-1, VCAM-1 and ET-1 in a concentration dependent manner or the repression of eNOS and NO. (2) Hydrolysis of sPLA2-IIA exactly take a part to regulate the expression of ICAM-1, VCAM-1, ET-1 and eNOS in endothelial cells. (3) sPLA2-IIA affect endothelial cells adhesion function primarily via ERK1/2 and NF-kB pathway. (4) sPLA2-IIA affect endothelial cells vasomotor function primarily via ERK1/2, JNK and NF-kB pathway.

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