OX40-OX40L相互作用对NFATc1表达及斑块形成的影响

The Effect of OX40-OX40L Interaction on the Expression of NFATc1 and Formation of the Atherosclerotic Plaques

目的探讨OX40-OX40L相互作用对活化T细胞核因子c1(NFATc1)及动脉粥样硬化斑块形成的影响。方法选取33只载脂蛋白(Apo)E-/-小鼠,分为对照组、OX40刺激组、OX40刺激+沉默NFATc1组,采用颈动脉硅胶圈置入法建立斑块模型;Masson染色检测斑块组成;采用免疫组化检测斑块内NFATc1和CD68分布。细胞实验以小鼠脑微静脉内皮细胞为对象,分为对照组、OX40刺激组和OX40抑制组。斑块及细胞表达NFATc1采用qRT-PCR和Western blot方法检测。结果细胞实验显示,OX40刺激组小鼠内皮细胞NFATc1蛋白及mRNA表达明显高于对照组(P<0.05),而OX40抑制组小鼠内皮细胞NFATc1蛋白及mRNA表达低于OX40刺激组(P<0.05)。动物实验显示,OX40刺激组Apo E-/-小鼠斑块中NFATc1蛋白及mRNA表达高于对照组(P<0.05),而OX40刺激+沉默NFATc1组Apo E-/-小鼠斑块中NFATc1蛋白及mRNA表达低于OX40刺激组(P<0.05)。OX40刺激组斑块面积与对照组相比显著增加,斑块内纤维增生明显,而OX40刺激+沉默NFATc1组较刺激组斑块面积明显减少,纤维增生程度减弱。OX40刺激组Apo E-/-小鼠斑块内NFATc1及CD68表达高于对照组,而OX40刺激+NFATc1沉默组小鼠斑块内NFATc1及CD68表达低于OX40刺激组。结论 OX40-OX40L信号调控NFATc1表达并影响动脉粥样硬化斑块的形成。

Aim To investigate the effect of OX40-OX40 L interaction on the expression of nuclear factor of activated T cells c1（NFATc1） and formation of the atherosclerotic plaques.Methods Atherosclerotic plaque model was randomly divided into the following groups： control group（n = 11）,OX40 activated group（n = 11） and OX40 activated＋PLKO.1-sh NFATc1 group（n = 11）.Atherosclerotic plaque model was produced by perivascular carotid collar placement in Apo E-/-mice.Masson staining was used to detect the composition of the plaque.Immunohistochemistry was used to observe the distribution of NFATc1 and CD68 in plaques.Mouse brain endothelial cells were divided into three groups：control group,OX40 activated group and OX40 inhibited group.The mRNA and protein level of NFATc1 was detected by real-time quantitative PCR（qRT-PCR） and Western blot respectively.Results In vitro,the expression level of NFATc1 was significantly increased in OX40 activated group,and were decreased in OX40 inhibited group（P0.05）.In vivo,the expression level of NFATc1 was significantly increased in OX40 activated group,and were decreased after the gene of NFATc1 being silenced（P0.05）.Masson staining revealed that the area of the plaque and fibrin proliferation was higher than the control group,while were decreased in OX40 activated＋PLKO.1-sh NFATc1 group.In vivo,compared with the control group,the distribution of NFATc1 and CD68 in the plaque were increased after the addition of agonist-OX40,which can be inhibited by silencing NFATc1.Conclusion OX40-OX40 L interaction may regulate the expression of NFATc1 and promote the formation of the atherosclerotic plaque.