Effects of γ-globulin on MMP-2, MMP-9 and TNF-αsecretion in human umbilical veins endothelial cells stimulated by serum from children with Kawasaki disease

Abstract

Abstract: AIM:To investigate the inducible matrix metalloproteases-2 (MMP-2), matrix metalloproteases- 9(MMP-9)and TNF-αsecretion in human umbilical veins endothelial cells (HUVECs)stimulated by the serum from children with acute phase of Kawasaki disease (KD)and the regulation effect of γ-globulin on MMP-2, MMP-9 and TNF-αsecretion. METHODS: HUVECs were cultured in six different conditional mediums with normal serum group (Group C), febrile serum group(Group F), KD without coronary artery lesion serum group(Group NAC), KD without coronary artery lesion serum added γ- globulin group (Group NACI), KD with coronary artery lesion serum group (Group AC), and KD with coronary artery lesion serum added γ-globulin group(Group ACI).The activities of MMP-2, MMP-9 and TNF-αin supernatant were detected using enzyme-linked immunosorbent assay (ELISA)after 12 h cultivation. RESULTS:The concentrations of MMP-2, MMP-9 and TNF-αin supernatant inHUVECs cultured with 10% serum from KD patients were significantly higher in Group AC or Group NAC than those in Group C and Group F (P <0. 01), there was statistical significance between in Group AC and Group NAC (P <0. 01).The concentrations of MMP-2, MMP-9 and TNF-αin supernatant in Group F were significantly lower than those in Group AC or Group NAC (P <0. 01), but were significantly higher than those in Group C (P <0. 01).The levels of MMP-2, MMP-9 and TNF-αin supernatant in HUVEC cultured with γ-globulin significantly were decreased in Group AC and Group NAC, there were statistical difference (P <0. 01).The levels of MMP-2 andMMP-9 in supernatant and the content of TNF-αhad significant positive correlation(r =0. 839, 0. 765, P <0. 01). CONCLUSION:The levels of MMP-2, MMP-9 and TNF-αin HUVEC induced by the serum f rom KD patients are increased and might take part in the formation and development of coronary artery lesions. The inhibition effects of γ-globulin on MMP-2, MMP-9 and TNF- αsecretion in endothelial cells could be related to the mechanism which γ-globulin prevents the happening of coronary artery of KD.

Key words: Kawasaki disease, γ-globulin, human umbilical veins endothelial cell, matrix metalloproteases-2, matrix metalloproteases-9, TNF-α

CLC Number:

R965. 2

ZHANG Yuan-hai, HE Yue-e, XIANG Ru-lian, WU Rong-zhou, XU Qiang, RONG Xing, CHEN Qi. Effects of γ-globulin on MMP-2, MMP-9 and TNF-αsecretion in human umbilical veins endothelial cells stimulated by serum from children with Kawasaki disease[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2009, 14(5): 586-590.

References

[1] Senzaki H.The pathophysiology of coronary artery aneury sms in kawasaki disease: role of matrix metalloproteinases[J]. Arch Dis Child, 2006, 91(10):847-851.
[2] Senzaki H, Masutani S, Kobayashi J, et al.Circulating matrix metalloproteinases and their inhibitors in patients with Kawasaki disease[J]. Circulation, 2001, 104(8): 860-863.
[3] Lau AC, Rosenberg H, Duong TT, et al. Elastolytic matrix metalloproteinases and coronary outcome in children with kawasaki disease[J]. Pediatr Res, 2007, 61(6):710^-715.
[4] Gavin PJ, Crawford SE, Shulman ST, et al.Systemic arterial expression of matrix metalloproteinases 2 and 9 in acute Kawasakidisease[J]. Arterioscler Thromb Vasc Biol, 2003, 23(4):576-581.
[5] Lau AC, Duong TT, Ito S, et al.Matrix metalloproteinase 9 activity leads to elastin breakdown in an animal model of Kawasaki disease[J]. Arthritis Rheum, 2008, 58(3):854-863.
[6] 张承俊, 朱兴雷.辛伐他汀对肿瘤坏死因子α诱导的人脐静脉内皮细胞基质金属蛋白酶9 表达的影响[J]. 中国动脉硬化杂志, 2006, 14(1):50-52.
[7] Wang BW, Chang H, Lin S. Induction of matrix metalloproteinases- 14 and-2 by cy clical mechanical stretch is mediated by tumor necrosis factor-alpha in cultured human umbilical vein endothelial cells [J]. Cardiovasc Res, 2003, 59(2):460-469.
[8] 陈春燕, 高玲, 陈琴, 等.重组C 反应蛋白促进脐静脉内皮细胞表达基质金属蛋白酶2[J]. 中华高血压杂志, 2007, 15(3):199-102.
[9] Hui Yuen JS, Duong TT, Yeung RS, et al. TNF-alpha is necessary for induction of coronary artery inflammation and aneurysm formation in an animal model of Kawasaki disease[J]. J Immunol, 2006, 76(10):6294-6301.
[10] Kaneko K, Savage CO, Pottinger BE, et al.Antiendothelial cell antibodies can be cytotoxic to endothelial cells without cytokine pre-stimulation and correlate with ELISA antibody measurement in kawasaki disease[J]. Clin Exp Immunol, 1994, 98(2):264-269.
[11] Sakata K, Kita M, Imanishi J, et al. Effect of Kawasaki disease on migration of human umbilical vein endothelial cells[J]. Pediatr Res, 1995, 38(4):501-505.
[12] Kazatchkine MD, Kaveri SV.Immunomodulation of autoimmune and inflammatory diseases with intravenous immune globulin[J]. N Engl J Med, 2001, 345(10):747-755.
[13] Makata H, Ichiyama T, Uchi R.Anti-inflammatory effect of intravenous immunoglobulin in comparison with dexamethasone in vitro: implication for treatment of Kawasaki disease [J]. Naunyn Schmiedebergs Arch Pharmacol, 2006, 373(5):325-332.
[14] Burns JC, Glode MP.Kawasaki syndrome[J]. Lancet, 2004, 364(9433):533-544.
[15] Stenbog EV, Windelborg B, Horlyck A, et al.The effect of TNFalpha blockade in complicated, refractory Kawasaki disease[J]. Scand J Rheumatol, 2006, 35(4):318-321.