细胞外基质金属蛋白酶诱导因子单克隆抗体抑制ApoE^((-/-))小鼠动脉粥样硬化形成的实验研究

Experimental study on the inhibitory effect of extracellular matrix metalloproteinase inducer monoclonal antibody on the formation of ApoE^((-/-)) atherosclerosis in mice

目的:探究EMMPRIN单克隆抗体对ApoE缺陷小鼠动脉粥样硬化(As)形成作用机制的实验研究。方法:选取健康的雄性,昆明小鼠10只和ApoE基因缺失ApoE^((-/-))小鼠30只,随机分为四组:空白对照组、EMMPRIN单克隆抗体干预组、ApoE^((-/-))As模型组、ApoE^((-/-))阴性对照组。利用ELASA试剂盒检测As模型动物外周血生化指标,观察EMMPRIN单克隆抗体对模型动物脂质代谢及As斑块的影响。利用RT-qPCR、Western-Blot和免疫组化等方法检测模型动物氧化应激信号通路中血管内皮生长因子(VEGF)和成纤维细胞生长因子2(FGP-2)的表达。结果:EMMPRIN单克隆抗体干预组与ApoE(-/-)阴性As模型组的比较中,在TG、TC、LDL-C等指标中,EMMPRIN单克隆抗体干预组明显低于ApoE(-/-)AS模型组,具有明显的统计学差异(t=4.327, 3.295, 1.773,P<0.05)。EMMPRIN单克隆抗体干预组的HDL-C均高于As模型对照组,具有明显的统计学差异(t=8.438,P<0.05)。②RT-PCR和Western blot结果显示,EMMPRIN单克隆抗体干预组中主动脉VEGF和FGP-2表达量明显低于As模型组,差异具有统计学意义(P<0.05)。③免疫组化显示,EMMPRIN单克隆抗体干预组中主动脉VEGF蛋白表达量与As模型组比较,差异有统计学意义(P<0.05).结论:EMMPRIN单克隆抗体通过抑制内质网应激VEGF蛋白和FGP-2蛋白的表达发挥其抑制As的作用。

Objective: an Experimental study on the effect of EMMPRIN Monoclonal Antibody on Atherosclerosis formation in ApoE deficient mice. Methods: a total of 10 healthy male Kunming mice and 30 ApoE^(-/-) mice were selected. They were randomly divided into 4 groups: the blank control group, EMMPRIN monoclonal antibody intervention group, Mouse atherosclerosis model with ApoE^(-/-) group, Negative control group with ApoE^(-/-). ELASA kit was used to detect peripheral blood biochemical markers in AS model animals. Observation of EMMPRIN monoclonal antibody on lipid metabolism and Atherosclerotic plaque in Model Animals. Expression of Vascular Endothelial growth Factor and fibroblast growth Factor 2 in oxidative stress signaling Pathway of Model Animals by RT-qPCRG Western-Blot and Immunohistochemistry. Results: In the comparison between the EMMPRIN monoclonal antibody intervention group and the ApoE^(-/-)negative atherosclerosis model group, the EMMPRIN monoclonal antibody intervention group was significantly lower than theApoE^(-/-) atherosclerosis model group in the TG、 TC、 LDL-C and so on. There was significant statistical difference between the EMMPRIN monoclonal antibody intervention group and the ApoE^(-/-) atherosclerosis model group(P<0.05). HDL-C in EMMPRIN monoclonal antibody intervention group was higher than that in atherosclerosis model control group(P<0.05).② The result of RT-PCR and Western blot showed that the expression of VEGF and FGP-2 in aorta was significantly lower in EMMPRIN monoclonal antibody intervention group than in atherosclerosis model group, with statistical difference(P<0.05).③Immunohistochemical staining showed that the expression of VEGF protein in aorta in the EMMPRIN monoclonal antibody intervention group was significantly different from that in the atherosclerosis model group(P<0.05). Conclusions: EMMPRIN monoclonal antibody inhibits atherosclerosis by inhibiting the expression of VEGF and FGP-2 proteins in endoplasmic reticulum stress.