miR-377介导Syk对ApoE-/-小鼠动脉粥样硬化病变的影响

miR-377 Mediates the effect of syk on atherosclerosis in apoE-/-mice

目的研究微小RNA-377(miR-377)介导脾酪氨酸激酶(Syk)对基因敲除(ApoE-/-)小鼠动脉粥样硬化病变的影响。方法将实验对象分为对照组(ApoE-/-小鼠)、miR-377组(注入miR-377的ApoE-/-小鼠)、Syk组(注入Syk的ApoE-/-小鼠)和Syk+miR-377组(注入miR-377和Syk的ApoE-/-小鼠)。通过制备miR-377和Syk质粒注入到ApoE-/-小鼠,通过qPCR和Western blot检测ApoE-/-小鼠Syk mRNA和蛋白的表达,检测炎症因子和脂质、氧化物歧化酶的水平。通过HE染色和油红O染色检测miR-377对ApoE-/-小鼠动脉粥样硬化主动脉斑块和表面积的影响。结果miR-377组的Syk mRNA相对表达明显低于对照组,miR-377组的Syk蛋白水平表达明显低于对照组。Syk组TNF-α水平明显低于对照组和miR-377组。miR-377组的1L-6水平明显高于对照组,Syk组和miR377+Syk组低于对照组。miR-377组的1L-1β水平明显低于对照组。miR-377组的TC水平明显低于对照组,Syk组的1L-1β水平高于对照组和miR-377组。miR-377组的LDL水平明显低于对照组,miR-377组的HDL水平明显低于对照组,Syk组的HDL水平明显高于对照组和miR-377组。miR-377组SOD的水平明显高于对照组。对照组、Syk组有明显的主动脉斑块,经过miR-377处理过的miR-377组和miR377+Syk组未见明显的主动脉斑块。miR-377处理的ApoE-/-小鼠,动脉粥样硬化斑块面积显著减少。syk处理的ApoE-/-小鼠斑块面积增加。对照组的硬化斑块面积/总面积之比为25.63%,miR-377组中为7.81%,Syk组为35.19%,Syk+miR-377组为9.73%。结论Syk基因的过表达会导致动脉粥样硬化细胞脂质的积累和氧化应激的增加,miR-377能够通过抑制Syk基因的表达来调控动脉粥样硬化的发展。

Objective To study the effect of miR-377-mediated Syk on atherosclerosis in ApoE/mice.Methods The experimental subjects were divided into control group(ApoE-/-mice),miR-377 group(ApoE-/-mice injected with miR-377),Syk group(ApoE-/-mice injected with Syk)and Syk+miR-377 group(ApoE-/-mice injected with miR-377and Syk).miR-377 single-stranded antisense nucleotide drugs and Syk gene sequences were prepared and transfected into ApoE-/-mice.The expression of Syk gene and protein in ApoE-/-mice was detected by qPCR and Western blot,and the levels of inflammatory factors,lipid and oxidative dismutase were detected.The effects of miR-377 on atherosclerotic aortic plaque and surface area in ApoE-/-mice were studied by HE staining and oil red O staining.Results The relative expression of Syk gene in miR-377 group was significantly lower than that in control group,and the expression of Syk protein in miR-377 group was significantly lower than that in control group.TNF-alpha levels in Syk group were significantly lower than those in control group and miR-377 group.The 1L-6 level in the miR-377 group was significantly higher than that in the control group,while the 1L-6 level in the Syk group and the MiR377+Syk group was significantly lower than that in the control group.The level of 1L-1βin the MiR377 group was significantly lower than that in the control group.The TC level of miR-377 group was significantly lower than that of control group,and the 1L-1βlevel of Syk group was significantly higher than that of control group and miR-377 group.The level of LDL in the miR-377 group was significantly lower than that in the control group,the level of HDL in the miR-377 group was significantly lower than that in the control group,and the level of HDL in the Syk group was significantly higher than that in the control group and the miR-377 group.The level of SOD in the miR-377 group was significantly higher than that in the control group.There were obvious aortic plaques in the control group and Syk group.There were no obvious aortic plaques in the miR-377+Syk group and the miR-377 treated group.The area of atherosclerotic plaque in ApoE-/-mice treated with miR-377 decreased significantly.The plaque area of ApoE-/-mice treated with Syk increased.The ratio of sclerotic plaque area to total area was 25.63%in control group,7.81%in miR-377 group,35.19%in Syk group and 9.73%in Syk+miR-377 group.ConclusionOverexpression of Syk gene can increase lipid accumulation and oxidative stress in atherosclerotic cells.miR-377 can regulate the development of atherosclerosis by inhibiting the expression of Syk gene.