Chen Xu-lin, Xia Zhao-fan, Wei Duo, Ben Dao-feng, Wang Yong-jie, Deng Nian-qing
Department of Burns, First Affiliated Hospital of Anhui Medical University, Hefei 230022, P.R. China.
Zhonghua Shao Shang Za Zhi. 2005 Dec;21(6):426-9.
To investigate the influence of burn serum on the expression of vascular cell adhesion molecule-1 (VCAM-1) of human umbilical vein endothelial cells (HUVECs) andits signal transduction mechanism.
HUVECs cultured in vitro were employed for the experiment, and were divided into normal control (NC, with addition of normal serum), burn serum (BS, with addition of burn serum), SB203580 (with addition of 10 micromol/L SB203580 treatment 1 hour before burn serum treatment) and PDTC [with 10 mmol/L pyrrolidine dithiocarbamate (PDTC) 1 hour before burn serum treatment] groups. Protein and mRNA expression of VCAM-1 in HUVECs was measured by flow cytometry and reverse transcription polymerase chain reaction (RT-PCR) respectively at 0, 6, 12, 24 and 36 hours after burn serum treatment. The expression of VCAM-1 on HUVEC surface and the soluble VCAM-1 (sVCAM-1) content in HUVECs culture supernatants were measured by ELISA at 24 hours after the serum stimulation. Adherence of peripheral blood mononuclear leukocytes (PBMC) adherence to HUVECs was also observed in vitro.
The expression of VCAM-1 mRNA increased obviously in BS group after the burn serum stimulation and reached peak level at 24 post stimulation hour (PSH), and it decreased thereafter. The above expression was significantly decreased in SB203580 and PDTC groups at 24 PSH, but there was no difference compared with normal control (P > 0.05). The VCAM-1 expression on the membrane of HUVEC was evidently higher in BS group (66.5 +/- 6.2) than that in NC group (19.1 +/- 1.9, P < 0.05) at 24 PSH, but it was decreased significantly in SB203580 (21.7 +/- 2.3) and PDTC (23.1 +/- 2.4) groups and there was no significant difference compared with NC group (P > 0.05), and which was evidently lower than that in BS group (P < 0.05). The VCAM-1 content in the supernatant of BS group (125 +/- 10 ng/L) was obviously higher than that in NC (23 +/- 3 ng/L), SB203580 (27 +/- 5 ng/L) and PDTC (29 +/- 5 ng/L) groups. (P < 0.05). The number of PBMCs adherent to HUVECs in BS group [(197 +/- 11)%] was much larger than that in NC group [(100 +/- 4)%], SB203580 group [(113 +/- 7)%] or PDTC group [(97 +/- 112)%] at 24 PSH (P < 0.05), but no difference between NC group and SB203580, PDTC groups (P > 0.05).
Burn serum can enhance the expression of VCAM-1 in HUVECs through p38 MAPK signaling pathway, and the activation of NF-kappaB was also involved in this process.
探讨烧伤血清对人脐静脉内皮细胞（HUVECs）血管细胞黏附分子-1（VCAM-1）表达的影响及其信号转导机制。
采用体外培养的HUVECs进行实验，分为正常对照组（NC，添加正常血清）、烧伤血清组（BS，添加烧伤血清）、SB203580组（在烧伤血清处理前1小时添加10 μmol/L SB203580处理）和PDTC组[在烧伤血清处理前1小时添加10 mmol/L吡咯烷二硫代氨基甲酸盐（PDTC）]。在烧伤血清处理后0、6、12、24和36小时，分别采用流式细胞术和逆转录聚合酶链反应（RT-PCR）检测HUVECs中VCAM-1的蛋白和mRNA表达。在血清刺激后24小时，采用酶联免疫吸附测定（ELISA）检测HUVECs表面VCAM-1的表达及HUVECs培养上清液中可溶性VCAM-1（sVCAM-1）的含量。体外观察外周血单个核白细胞（PBMC）对HUVECs的黏附情况。
烧伤血清刺激后，BS组VCAM-1 mRNA表达明显增加，在刺激后24小时达到峰值水平，此后下降。在刺激后24小时，SB203580组和PDTC组上述表达明显降低，但与正常对照组相比无差异（P>0.05）。在刺激后24小时，BS组HUVECs膜上VCAM-1表达明显高于NC组（66.5±6.2比19.1±1.9，P<0.05），但在SB203580组（21.7±2.3）和PDTC组（23.1±2.4）明显降低，与NC组相比无显著差异（P>0.05），且明显低于BS组（P<0.05）。BS组上清液中VCAM-1含量（125±10 ng/L）明显高于NC组（23±3 ng/L）、SB203580组（27±5 ng/L）和PDTC组（29±5 ng/L）（P<0.05）。在刺激后24小时，BS组黏附于HUVECs的PBMC数量[（197±11）%]明显多于NC组[（100±4）%]、SB203580组[（113±7）%]或PDTC组[（97±112）%]（P<0.05），但NC组与SB203580组、PDTC组之间无差异（P>0.05）。
烧伤血清可通过p38丝裂原活化蛋白激酶（MAPK）信号通路增强HUVECs中VCAM-1的表达，核因子κB（NF-κB）的激活也参与此过程。
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