Yin Jun, Luo Xin Guo, Yu Wen Jun, Liao Jing Yuan, Shen You Jin, Zhang Ze Wen
Division of Hematology, the Second Hospital affiliated to Medical College of Shantou University, Shantou, China.
Cell Physiol Biochem. 2010;25(4-5):477-90. doi: 10.1159/000303053. Epub 2010 Mar 23.
The role of antisense oligodeoxynucleotide against tissue factor (aODN/TF) in cultured human umbilical vein endothelial cells (HUVECs) subjected to anoxia-reoxygenation (A/R) was investigated.
HUVECs were divided randomly into control group, A/R group, aODN/TF+A/R group, sense oligodeoxynucleotide (sODN/TF) + A/R group and mismatched oligodeoxynucleotide (mODN/TF) + A/R group, in the latter 3 groups, HUVECs were transfected with aODN/TF, sODN/TF and mODN/TF respectively. HUVECs in all A/R groups underwent 3 hrs of anoxia and followed by 2 hrs of reoxygenation. In order to investigate the potential mechanisms of how increased TF may contribute to A/R injury in HUVECs, another set of HUVECs were incubated with human recombinant active site blocked factor VII (FVIIai) during A/R.
After A/R, TF expression at both mRNA and protein level was increased, furthermore, cell viability and the concentrations of SOD, GSH-PX and NO were declined, while LDH, MDA and ET-1 were overproduced (P<0.05 to 0.001 versus control group). In HUVECs of aODN/TF+A/R group, however, TF expression was inhibited, while the declined cell viability and the concentrations of SOD, GSH-PX, NO as well as the enhanced LDH, MDA and ET-1 levels occurred during A/R were ameliorated and reversed effectively (P<0.05 to 0.01 versus those in other A/R groups). The results also showed that ROS was increased and PAR-1, PAR-2, p38 MAP kinase and p42/44 MAP kinase were all activated after A/R (P<0.001 versus HUVECs under normoxia), while FVIIai inhibited the increment of ROS, PAR-1, PAR-2, p38 MAP kinase and p42/44 MAP kinase, and improved the changes of TF:C, MDA, SOD, GSH-PX, cell viability and LDH occurred during A/R (P<0.05 to 0.001 versus HUVECs without FVIIai treatment).
Tissue factor plays an important role in the development of HUVECs injury induced by anoxia-reoxygenation, inhibition of TF with antisense oligodeoxynucleotide is an effective approach to ameliorate the damage.
研究反义寡脱氧核苷酸靶向组织因子（aODN/TF）在缺氧复氧（A/R）处理的人脐静脉内皮细胞（HUVECs）中的作用。
将HUVECs随机分为对照组、A/R组、aODN/TF + A/R组、正义寡脱氧核苷酸（sODN/TF）+ A/R组和错配寡脱氧核苷酸（mODN/TF）+ A/R组，后3组分别用aODN/TF、sODN/TF和mODN/TF转染HUVECs。所有A/R组的HUVECs均经历3小时缺氧，随后2小时复氧。为了研究组织因子增加如何导致HUVECs发生A/R损伤的潜在机制，另一组HUVECs在A/R期间与人重组活性位点阻断因子VII（FVIIai）孵育。
A/R后，组织因子在mRNA和蛋白水平的表达均增加，此外，细胞活力以及超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-PX）和一氧化氮（NO）的浓度下降，而乳酸脱氢酶（LDH）、丙二醛（MDA）和内皮素-1（ET-1）产生过多（与对照组相比，P<0.05至0.001）。然而，在aODN/TF + A/R组的HUVECs中，组织因子表达受到抑制，同时A/R期间出现的细胞活力下降以及SOD、GSH-PX、NO浓度降低以及LDH、MDA和ET-1水平升高均得到有效改善和逆转（与其他A/R组相比，P<0.05至0.01）。结果还表明，A/R后活性氧（ROS）增加，蛋白酶激活受体-1（PAR-1）、蛋白酶激活受体-2（PAR-2）、p38丝裂原活化蛋白激酶（p38 MAP激酶）和p42/44丝裂原活化蛋白激酶均被激活（与常氧下的HUVECs相比，P<0.001），而FVIIai抑制了ROS、PAR-1、PAR-2、p38 MAP激酶和p42/44 MAP激酶的增加，并改善了A/R期间发生的组织因子促凝活性（TF:C）、MDA、SOD、GSH-PX、细胞活力和LDH的变化（与未用FVIIai处理的HUVECs相比，P<0.05至0.001）。
组织因子在缺氧复氧诱导的HUVECs损伤发展中起重要作用，用反义寡脱氧核苷酸抑制组织因子是改善损伤的有效方法。
