蘘荷氯仿提取物对人真皮微血管内皮细胞表面黏附分子的表达及淋巴细胞黏附作用的影响

【摘要】 目的 探讨蘘荷对人真皮微血管内皮细胞（HDMEC）表面黏附分子的表达及淋巴细胞黏附作用的影响。方法 用蘘荷氯仿提取物（CFMG）与肿瘤坏死因子（TNF）-α/白介素（IL）-1α分别诱导或按不同顺序联合诱导HDMEC不同时间，用酶联免疫吸附试验（ELISA）检测细胞间黏附分子-1（ICAM-1）、血管细胞间黏附分子-1（VCAM-1）和E选择素表达水平。使用γ计数器测定HDMEC对T淋巴细胞和Ramos细胞的黏附能力。用Sigma Plot 12软件包对数据进行配对t检验。 结果 与0.2%二甲基亚砜相比，CFMG轻度下调HDMEC表面ICAM-1、VCAM-1和E选择素表达水平（均P > 0.05）；与细胞培养液相比，TNF-α显著上调ICAM-1、VCAM-1、E选择素的表达水平（均P < 0.01），IL-1α上调ICAM-1和E选择素的表达水平（均P < 0.01），对VCAM-1轻度上调（P > 0.05）。CFMG预处理可显著抑制TNF-α和IL-1α对黏附分子表达水平的上调作用（均P < 0.01），但是CFMG后处理对经TNF-α和IL-1α刺激后升高的黏附分子表达水平影响不大（均P > 0.05）。与0.2% DMSO相比，CFMG轻度降低HDMEC对T淋巴细胞和Ramos细胞黏附率（均P > 0.05），而与细胞培养液相比，TNF-α和IL-1α显著提高其黏附率（均P < 0.01）。 结论 蘘荷脂溶性粗提物可能通过阻遏前炎症细胞因子TNF-α和IL-1α对HDMEC表面黏附分子的上调作用，抑制组织中炎细胞浸润从而起到抗炎作用。 【关键词】 蘘荷； 内皮细胞； 细胞黏附分子； 细胞因子

Effect of chloroform extract of Zingiber mioga Roscoe on human dermal microvascular endothelial cell expression of cell adhesion molecules and adhesivity to lymphocytes

LI Zhong-zhu, ZHANG Qi-guo*, LI Wei-hua, SHI Song-lin, LI Qi-fu. *Department of Dermatology, First Affiliated Hospital of Xiamen University, Xiamen 361003, China Corresponding author: LI Zhong-zhu, Email: leezhongzhu@hotmail.com 【Abstract】 Objective To investigate the effect of chloroform extract of Zingiber mioga Roscoe on human dermal microvascular endothelial cell （HDMEC） expression of cell adhesion molecules and adhesivity to lymphocytes. Methods HDMECs were isolated from human foreskin tissue and subjected to subculture. After several passages, some HDMECs were treated with chloroform extract from Zingiber mioga Roscoe （CFMG） of 200 mg/L for 30 minutes before or after 24-hour stimulation with tumor necrosis factor （TNF）-α or interleukin （IL）-1α. Subsequently, enzyme-linked immunosorbent assay was performed to determine the expression levels of intercellular adhesion molecule-1 （ICAM-1）, vascular cell adhesion molecule-1 （VCAM-1） and E-selectin on HDMECs. Both T lymphocytes isolated from venous blood of healthy adults and Ramos Burkitt′s lymphoma B cells were labelled with chromium-51 and incubated with the HDMECs for four hours followed by the determination of radiation intensity of lymphocytes adhering to HDMECs using a gamma-counter. Results Compared with 0.2% dimethyl sulfoxide （DMSO）, CFMG slightly down-regulated the expression of ICAM-1, VCAM-1 and E-selectin on resting HDMECs （all P > 0.05）. In comparison with culture medium, TNF-α enhanced the expressions of ICAM-1, VCAM-1 and E-selectin significantly （all P < 0.01）, and IL-1α elevated the expressions of ICAM-1 and E-selectin markedly（both P < 0.01） as well as the expression of VCAM-1 slightly （P > 0.05）. The upregulating effects of TNF-α and IL-1α on the expressions of adhesion molecules were notably suppressed by the CFMG treatment prior to the stimulation with TNF-α and IL-1α（all P < 0.01）, but not affected by that after the stimulation （all P > 0.05）. The adhesivity of HDMECs to T lymphocytes and Ramos cells was slightly decreased by CFMG treatment compared with 0.2% DMSO（both P > 0.05）, but was markedly increased by TNF-α and IL-1α compared with the culture medium （both P < 0.01）. Conclusions CFMG may play an anti-inflammatory role via blocking the up-regulating effect of pre-inflammatory cytokines such as TNF-α and IL-1α on the expression of adherence molecules on HDMECs and, hence, inhibiting inflammatory cell infiltration in tissue. 【Key words】 Zingiber mioga Roscoe; Endothelial cells; Cell adhesion molecules; Cytokines