Effects of miR-125a-3p on Atherosclerotic Plaque, M1/M2 Macrophages, MMP-9 and VEGF
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摘要:
目的 探讨miR-125a-3p对大耳兔动脉粥样硬化斑块形成、M1/M2巨噬细胞、基质金属蛋白酶9（MMP-9）和血管内皮生长因子（VEGF）的影响。 方法 15只成年雄性健康日本大耳兔，随机分为3组，每组5只，对照组给予普通饲料喂养，动脉粥样硬化模型组给予牛血清白蛋白注射和高胆固醇饲料喂养，miR-125a-3p抑制剂干预组给予动脉粥样硬化兔注射miR-125a-3p干扰慢病毒载体。油红O染色后图像分析法测定斑块面积，免疫组化法测定MMP-9和VEGF，免疫荧光法检测斑块组织中M1标记物CD11c和M2标记物CD206。 结果 与动脉粥样硬化组相比，miR-125a-3p抑制剂组斑块面积百分比显著降低（P < 0.0001）。与对照组相比，动脉粥样硬化组MMP-9（P < 0.001）、VEGF（P < 0.01）和CD11c（P < 0.001）水平显著升高，CD206水平显著降低（P < 0.001）；与动脉粥样硬化组相比，miR-125a-3p抑制剂干预组MMP-9（P < 0.01）、VEGF（P < 0.01）和CD11c（P < 0.001）水平有所降低，CD206水平有所升高（P < 0.001）。 结论 抑制miR-125a-3p可减轻动脉粥样硬化斑块的形成，平衡M1/M2巨噬细胞，降低斑块组织中MMP-9和VEGF的表达，miR-125a-3p可能是治疗不稳定动脉粥样硬化斑块的新靶点。 -
关键词:
- miR-125a-3p抑制剂 /
- 动脉粥样硬化 /
- M1/M2巨噬细胞 /
- 基质金属蛋白酶9 /
- 血管内皮生长因子
Abstract:Objective To investigate the effects of miR-125a-3p inhibitors on the atherosclerotic plaque formation, M1/M2 macrophages, MMP-9 and VEGF. Methods Fifteen healthy adult male Japanese big-eared rabbits were randomly divided into 3 groups with 5 rabbits in each group. Those in the control group were fed with the normal diet,, those in the atherosclerosis model group were injected with bovine serum albumin and fed with the high cholesterol diet, and those in the miR-125a-3p inhibitor intervention group were injected with miR-125a-3p to interfere with lentivirus vector. The plaque area was determined by image analysis after oil red O staining, MMP-9 and VEGF were determined by immunohistochemistry, and M1 marker CD11c and M2 marker CD206 in plaque tissues were detected by immunofluorescence. Results Compared with the atherosclerosis group, the percentage of plaque area in miR-125a-3p inhibitor group decreased significantly （P < 0.0001）. Compared with the control group, the levels of MMP-9 （P < 0.001）, VEGF （P < 0.01） and CD11c （P < 0.001） increased significantly, and the level of cd206 decreased significantly in the atherosclerosis group （P < 0.001）; Compared with the atherosclerosis group, the levels of MMP-9 （P < 0.01）, VEGF （P < 0.01） and CD11c （P < 0.001） decreased, and the level of cd206 increased in miR-125a-3p inhibitor intervention group （P < 0.001）. Conclusion Inhibition of miR-125a-3p can reduce the formation of atherosclerotic plaques, balance M1/M2 macrophages, and reduce the expression of MMP-9 and VEGF in plaque tissues.miR-125a-3p may be a new target for the treatment of unstable plaques. -
Key words:
- miR-125a-3p inhibitors /
- Atherosclerosis /
- M1/M2 macrophages /
- MMP-9 /
- VEGF
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图 1 血管内膜油红“O”染色各组动脉粥样硬化病变区域
与对照组比较，****P < 0.0001，****P < 0.0001；与AS模型组比较，***P < 0.001。
Figure 1. Vascular intima oil red "O" staining for atherosclerotic lesions in each group
图 2 血管内膜油红“O”染色
a：对照组；b：AS模型组；c：AS模型+miR-125a-3p抑制剂干预组；miR-125a-3p抑制剂干预后动脉粥样硬化组病变区域显著减小。
Figure 2. Vascular intima is stained with oil red "O" stain
图 3 免疫组化检测各组MMP-9 表达水平
与对照组比较，***P < 0.001，*P < 0.05；与AS模型组比较，**P < 0.01。
Figure 3. The expression level of MMP-9 was detected by immunohistochemistry
图 4 MMP-9免疫组化染色（×200）
a：对照组；b：AS模型组；c：miR-125a-3p抑制剂干预组；miR-125a-3p抑制剂干预后，MMP-9表达水平显著下降。
Figure 4. MMP-9 immunohistochemical staining （×200）
图 5 免疫组化检测各组VEGF表达水平
与对照组比较，**P < 0.01，*P < 0.05；与AS模型组比较，**P < 0.01。
Figure 5. Immunohistochemistry was used to detect VEGF expression levels in each group
图 6 VEGF免疫组化染色（×200）
a：对照组；b：AS模型组；c：AS模型+miR-125a-3p抑制剂干预组；miR-125a-3p抑制剂干预后，VEGF表达水平下降。
Figure 6. VEGF immunohistochemical staining （×200）
图 7 免疫荧光检测各组M1标志物CD11c表达水平
与对照组比较，****P < 0.0001，*P < 0.05；与AS模型组比较，****P < 0.0001。
Figure 7. The expression level of M1 marker CD11c was detected by immunofluorescence
图 8 M1标志物CD11c免疫荧光染色（×200）
a：对照组；b：AS模型组；c：AS模型+miR-125a-3p抑制剂干预组；miR-125a-3p抑制剂干预后，CD11c表达水平显著下降。
Figure 8. M1 marker CD11c immunofluorescence staining （×200）
图 9 免疫荧光检测各组M2 标志物CD206表达水平
与对照组比较，****P < 0.0001，**P < 0.01；与AS模型组比较，****P < 0.0001。
Figure 9. The expression level of M2 marker CD206 was detected by immunofluorescence
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