疡愈涂剂对人白细胞与血管内皮细胞黏附影响的分子机制

Effects and Molecular Mechanisms of YYTJ on the Adhesion of Leukocytes to Human Umbilical Vein Endothelial Cells

目的研究疡愈涂剂(YangYuTuJi,YYTJ)对白细胞与血管内皮细胞黏附影响的分子机制,探讨其促进皮肤创伤愈合的机制。方法用虎红染色法观察YYTJ高(132.5mg/L)、中(66.3mg/L)、低(33.2mg/L)浓度对中性粒细胞(polymorphonuclear,PMN)、人单核细胞(THP-1)与肿瘤坏死因子(tumor necrosis factor,TNF)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)黏附的影响,用荧光免疫细胞化学法观察YYTJ对HUVEC表面细胞间黏附分子-1(inter cell ularadhesion molecule-1,ICAM-1)、血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)和CD44表达的影响。结果 YYTJ和TNF共同作用HUVEC12h后,高浓度YYTJ能明显抑制HUVEC与PMN以及THP-1黏附(P<0.05),能明显抑制HUVEC表面ICAM-1和CD44表达(P<0.05)。中浓度YYTJ能明显抑制HUVEC表面VCAM-1和CD44表达(P<0.05,P<0.01)。低浓度YYTJ能明显降低HUVEC表面CD44表达(P<0.05)。结论疡愈涂剂通过降低TNF诱导的HUVEC表面黏附分子的表达,抑制白细胞与血管内皮细胞的黏附,从而抑制过度的炎性反应。

Objective To observe the effects and molecular mechanisms of a traditional Chinese medicine formula Yang Yu Tu Ji（YYTJ） on the adhesion of leukocytes to human umbilical vein endothelial cells（HUVEC）,and study the mechanisms of YYTJ in promoting skin wound healing.Methods YYTJ at 132.5 μg/mL,which was confirmed to have no toxic effect on cells activity was selected as the high dose to observe the effect on HUVEC and human peripheral polymorphonuclear（PMN） leukocyte activity by MTT and Trypan-blue assay respectively.The effect of YYTJ at high（132.5 mg/L）,medium（66.3 mg/L）and low（33.2 mg/L）concentration on the adhesion of PMN/THP-1 to HUVEC primed by TNF were observed by Rose Bengal staining.The expression of ICAM-1,VCAM-1 and CD44 on HUVEC were measured by fluorescent-immunocytochemistry assay.Results High concentration of YYTJ reduced PMN/THP-1-HUVEC adhesion at 12 h after treating with YYTJ and TNF,showed lower level of ICAM-1 and CD44 on HUVEC（P0.05）.Medium dose of YYTJ decreased the level of VCAM-1 and CD44 on HUVEC（P0.05,P0.01 respectively）.Low dose of YYTJ inhibited the level of CD44 on HUVEC（P0.05）.Conclusion It’s possible that YYTJ inhibit adhesion of leukocytes to HUVEC by decreasing the expression of adhesion molecules on HUVEC,therefore inhibit excessive inflammatory response.