Sestrin2蛋白对动脉粥样硬化的干预效果及作用机制研究

Intervention effect and mechanism of Sestrin2 protein on atherosclerosis

目的探讨Sestrin2蛋白对动脉粥样硬化病变干预效果及作用机制。方法选择30只雄性ApoE-/-小鼠,建立动脉粥样硬化模型。模型建立后分为3组:9只小鼠经慢病毒处理(沉默组,Sestrin2基因沉默,原计划该组纳入10只小鼠,因其中1只处理后出现病态表现遂予以剔除),10只经过表达Sestrin2腺病毒处理(过表达组),余10只为对照组。分组处理完毕后20 d检测动脉粥样硬化斑块相关指标:斑块面积、最小管腔直径、管腔面积、白细胞介素(IL)-1β、肿瘤坏死因子α(TNF-α)、IL-10、基质金属蛋白酶(MMP)8、MMP12、MMP13,以及与Sestrin2蛋白表达相关联的磷酸化c-Jun氨基末端激酶(p-JNK)和磷酸化c-Jun(p-c-Jun)表达水平。结果对照组管腔直径、管腔面积大于沉默组(P<0.05),但均低于过表达组(P<0.05);沉默组斑块面积明显大于对照组和过表达组(P<0.01),且过表达组明显小于对照组(P<0.01)。沉默组IL-1β、TNF-α水平明显高于对照组和过表达组(P<0.01),而过表达组明显低于对照组(P<0.05);沉默组IL-10水平明显低于对照组和过表达组(P<0.01),而过表达组明显高于对照组(P<0.01)。沉默组MMP8、MMP12及MMP13水平高于对照组和过表达组(P<0.05),而过表达组又明显低于对照组(P<0.01)。过表达组p-JNK蛋白表达水平明显高于对照组和沉默组(P<0.01),而对照组表达水平又高于沉默组(P<0.01);各组小鼠p-c-Jun蛋白表达水平比较差异均无统计学意义(P>0.05)。结论Sestrin2蛋白通过调控MMP、IL等因子表达水平抑制动脉粥样硬化病变过程。

Objective To investigate the intervention effect and mechanism of Sestrin2 protein on atherosclerotic lesion.Methods Thirty male ApoE-/-mice were selected to establish the atherosclerosis model.After constructing the model,the mice were divided into the three groups:9 mice were treated with lentivirus(silencing group,Sestrin2 gene silencing,10 mice were originally included in this group,but 1 mouse was removed due to appear the sickness after treatment),10 mice were treated with Sestrin2 expressed adenovirus method(overexpression group),and the remaining 10 mice served as the control group.On 20 d after treatment end,the atherosclerotic plaque related indicators:plaque area,minimal lumen diameter,lumen area,interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),matrix metalloproteinase 8(MMP 8),MMP12,MMP13,p-JNK and phosphorilation p-c-Jun expression levels were detected.Results The lumen diameter and lumen area of the control group were larger than those of the silencing group(P<0.05),but lower than those of the overexpression group(P<0.05).The plaque area of the silencing group was significantly larger than that of the control group and the overexpression group(P<0.01),moreover the overexpression group was significantly smaller than that of the control group(P<0.01).The levels of IL-1βand TNF-αin the silencing group were significantly higher than those in the control group and the overexpression group(P<0.01),while the overexpression group was significantly lower than the control group(P<0.05).The level of IL-10 in the silencing group was significantly lower than that in the control group and the overexpression group(P<0.01),while the overexpression group was significantly higher than the control group(P<0.01).The levels of MMP8,MMP12 and MMP13 in the silencing group were higher than those in the control group and the overexpression group(P<0.05),while the overexpression group was significantly lower than the control group(P<0.01).The expression level of p-JNK protein in the overexpressed group was significantly higher than that in the control group and the silencing group(P<0.01),while the expression level in the control group was higher than that in the silencing group(P<0.01).There was no statistically significant difference in protein expression level of p-c-Jun among various groups(P>0.05).Conclusion Sestrin2 protein inhibits the atherosclerotic lesion process by regulating the expression levels of the factors such as MMP and interleukin,thus may inhibit the atherosclerosis lesion process.