川崎病急性期血清对血管内皮MMP-2、 MMP-9和TNF-α分泌的影响及IVIG 的干预机制探讨

摘要/Abstract

摘要： 目的:观察川崎病(Kawasaki disease, KD)患儿急性期血清对脐静脉内皮细胞(HUVEC)分泌基质金属蛋白酶-2(matrix metalloproteases-2, MMP-2)、基质金属蛋白酶-9(matrix metalloproteases-9, MMP-9)和TNF-α的影响及人血丙种球蛋白(IVIG)对上述过程的调节作用。方法:HUVEC 培养分为正常血清组(C 组)、发热血清组(F 组)、川崎病冠脉无损害血清组(NAC 组)、川崎病冠脉无损害血清+IVIG 组(NACI 组)、川崎病冠脉损害血清组(AC 组)、川崎病冠脉损害血清+IVIG 组(ACI 组)。每组分别培养12 h 后吸取上清液, 应用ELISA 法测定细胞上清液MMP-2、 MMP-9 和TNF-α水平。结果:经川崎病患儿血清作用后的HUVEC 培养上清液中MMP-2、 MMP-9 和TNF-α水平升高明显, 与C 组、F 组相比差异有统计学意义(P 均<0. 01), 其中AC 组更高, 与NAC 组相比差异有统计学意义(P <0. 01);F 组上清液中MMP-2、 MMP-9 和TNF-α水平虽然低于川崎病组, 但仍高于C 组(P <0. 01)。经丙种球蛋白干预后,AC组、NAC 组MMP-2、 MMP-9 和TNF-α水平下降, 差异有统计学意义(P 均<0. 01)。HUVEC 培养上清液中MMP-2、 MMP-9 与TNF-α含量之间呈显著性正相关(r =0. 839, 0. 765, P <0. 01)。结论:川崎病患儿急性期血清能诱导HUVEC 分泌MMP-2、 MMP-9 和TNF-α增加,MMP-2、 MMP-9 和TNF-α可能参与川崎病患儿冠脉损害的发生发展;IVIG 防治冠脉损害的机制, 可能与抑制内皮细胞分泌MMP-2、 MMP-9 和TNF-α有关。

关键词: 川崎病, 人血丙种球蛋白, 脐静脉内皮细胞, 基质金属蛋白酶-2, 基质金属蛋白酶-9, TNF-α

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

中图分类号:

R965. 2

张园海, 何跃娥, 项如莲, 吴蓉洲, 徐强, 荣星, 陈其. 川崎病急性期血清对血管内皮MMP-2、 MMP-9和TNF-α分泌的影响及IVIG 的干预机制探讨[J]. 中国临床药理学与治疗学, 2009, 14(5): 586-590.

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.

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