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[摘要]
目的 采用实时荧光定量PCR(qRT-PCR)法探索全反式维甲酸(ATRA)对兔颈动脉粥样硬化斑块组织炎症因子表达的影响。方法 选取24只纯种雄性新西兰白兔随机分为对照组、模型组、ATRA组。对照组给予基础饲料、其余两组给予高脂饲料,共饲养12周。饲养4周后,ATRA组ig 5 mg/kg的ATRA,模型组和对照组ig等量溶剂,每天1次,至12周末处死各组动物。取颈动脉粥样硬化斑块组织,进行HE染色,显微镜下观察管壁结构;提取组织RNA,qRT-PCR法测定组织中转化生长因子-β1(TGF-β1)、白细胞介素-10(IL-10)、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9) mRNA的相对表达量。结果 模型组可见典型的AS斑块,可见薄层的纤维帽及大的脂质核心,以及大量炎症细胞浸润,内弹力纤维膜断裂;ATRA组内皮细胞形态基本完整,可见少量炎性细胞浸润。与对照组比较,模型组中TGF-β1和IL-10 mRNA的相对表达量明显降低(P<0.05),而MMP-2和MMP-9 mRNA的相对表达量明显升高(P<0.05);与模型组比较,ATRA组的TGF-β1和IL-10 mRNA的相对表达量明显升高(P<0.05),MMP-2和MMP-9 mRNA的相对表达量明显降低(P<0.05)。结论 ATRA可通过调节炎症因子的表达抗动脉粥样硬化。
[Key word]
[Abstract]
Objective To investigate the effects of all-trans retinoic acid (ATRA) on the inflammatory response in carotid atherosclerotic plaques of rabbit model by using fluorescent quantitative PCR. Methods Twenty-four white purebred male New Zealand rabbits were randomly divided into control group, model group, and ATRA group. The rabbits in the control group were treated with basal feed, while the other two groups were fed with high fat diet. Four weeks later, ATRA group received ATRA ravage at a 5 mg dose once a day and model group received saline ravage at the same dose once a day. After 12 weeks, all rabbits were sacrificed and the carotid atherosclerotic plaques of rabbit were extracted. HE staining was performed and the wall structure was observed under microscope. With the mRNA of GAPDH as internal reference, the relative mRNA expression of TGF-β1, IL-10, MMP-2, and MMP-9 were detected by qRT-PCR. Results The typical AS plaque, thin fibrous cap and large lipid core, infiltration of inflammatory cells and breakage of elastic fibrous membrane were observed in the model group. The endothelial cells in the ATRA group were basically intact with a small amount of inflammatory cells infiltration. Compared with control group, the relative mRNA expression of TGF-β1 and IL-10 were decreased obviously in model group (P < 0.05), while MMP-2 and MMP-9 mRNA were increased significantly (P < 0.05).Compared with model group, the relative mRNA expression of TGF-β1 and IL-10 were increased significantly in ATRA group (P < 0.05), while MMP-2 and MMP-9 mRNA decreased obviously (P < 0.05). Conclusion ATRA can exert anti-atherosclerosis effect by regulating the expression of inflammatory factors.
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[基金项目]
广东药学院科技处-附属第一医院联合自然科学培育基金项目(GYFYLH201322);广州市越秀区科技计划项目(2017-WS-010)