CAJ | 학술논문

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-10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.
목적 탐토양하대인진피미혈관내피세포(HDMEC)표면점부분자적표체급림파세포점부작용적영향.방법 용양하록방제취물(CFMG)여종류배사인자(TNF)-α/백개소(IL)-1α분별유도혹안불동순서연합유도HDMEC불동시간,용매련면역흡부시험(ELISA)검측세포간점부분자-1(ICAM-1)、혈관세포간점부분자-1(VCAM-1)화E선택소표체수평.사용γ계수기측정HDMEC대T림파세포화Ramos세포적점부능력.용Sigma Plot 12연건포대수거진행배대t검험.결과 여0.2％이갑기아풍상비,CFMG경도하조HDMEC표면ICAM-1、VCAM-1화E선택소표체수평(균P＞0.05)；여세포배양액상비,TNF-α현저상조ICAM-1、VCAM-1、E선택소적표체수평(균P＜0.01),IL-1α상조ICAM-1화E선택소적표체수평(균P＜ 0.01),대VCAM-1경도상조(P＞0.05).CFMG예처리가현저억제TNF-α화IL-1α대점부분자표체수평적상조작용(균P＜ 0.01),단시CFMG후처리대경TNF-α화IL-1α자격후승고적점부분자표체수평영향불대(균P＞ 0.05).여0.2％ DMSO상비,CFMG경도강저HDMEC대T림파세포화Ramos세포점부솔(균P＞ 0.05),이여세포배양액상비,TNF-α화IL-1α현저제고기점부솔(균P＜0.01).결론 양하지용성조제물가능통과조알전염증세포인자TNF-α화IL-10대HDMEC표면점부분자적상조작용,억제조직중염세포침윤종이기도항염작용.
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 antiinflammatory 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.