叶酸联合维生素B12对高同型半胱氨酸血症引起大鼠动脉粥样硬化的治疗作用

doi:10.13481/j.1671-587x.20170516

摘要/Abstract

摘要： 目的：探讨叶酸联合维生素B12对高同型半胱氨酸血症引起大鼠动脉粥样硬化的治疗作用，并阐明其治疗机制。方法：将33只大鼠随机分为对照组、模型组和治疗组，每组11只，对照组大鼠给予普通饲料，模型组大鼠给予普通饲料和蛋氨酸，治疗组大鼠给予普通饲料和蛋氨酸同时灌胃给予叶酸和维生素B12。HE染色观察大鼠胸主动脉的病理组织形态表现，ELISA法检测大鼠血清同型半脱氨酸（Hcy）和基质金属蛋白酶9（MMP-9）水平，免疫组织化学染色检测大鼠胸主动脉平滑肌细胞中CC趋化因子受体5（CCR5）和CC趋化因子配体5（CCL5）蛋白表达水平，RT-PCR法检测胸主动脉平滑肌细胞中CCR5和CCL5 mRNA表达水平。结果：HE染色，对照组大鼠胸主动脉中膜平滑肌细胞排列整齐，内弹力膜完整，内皮细胞连续；模型组大鼠胸主动脉中膜平滑肌细胞紊乱疏松，内膜损伤，内膜下有泡沫细胞形成，内皮细胞坏死脱落；治疗组大鼠胸主动脉病理组织形态表现介于对照组和模型组之间。模型组和治疗组大鼠血清Hcy水平高于对照组（P < 0.05），治疗组大鼠血清Hcy水平低于模型组（P < 0.05）；模型组和治疗组大鼠血清MMP-9水平高于对照组（P < 0.05），治疗组大鼠血清MMP-9水平低于模型组（P < 0.05）；模型组和治疗组大鼠胸主动脉平滑肌细胞中CCR5和CCL5蛋白灰度值低于对照组（P < 0.05），治疗组大鼠胸主动脉平滑肌细胞中CCR5和CCL5蛋白表达水平高于模型组（P < 0.05）；模型组和治疗组大鼠胸主动脉平滑肌细胞中CCR5和CCL5 mRNA相对表达水平高于对照组（P < 0.05），治疗组大鼠胸主动脉平滑肌细胞中CCR5和CCL5 mRNA相对表达水平低于模型组（P < 0.05）。结论：叶酸联合维生素B12治疗可以降低血清Hcy和MMP-9水平和胸主动脉平滑肌细胞中CCR5及CCL5表达水平，这可能是叶酸联合维生素B12治疗动脉粥样硬化机制之一。

关键词: 动脉粥样硬化, 叶酸, 维生素B12, 基质金属蛋白酶9, CC型趋化因子受体5, CC型趋化性细胞因子5

Abstract: Objective: To investigate the therapeutic effect of folic acid combined with vitamin B12 on atherosclerosis induced by hyperhomocysteinemia in the rats, and to elucidate its therapeutic mechanism. Methods: Thirty-three rats were randomly divided into control group, model group and treatment group (n=11).The rats in control group were given normal diet,the rats in model group were given normal diet and methionine,and the rats in treatment group were given normal diet, methionine, perfused folic acid and vitamin B12. The histopathological changes of thoracic aorta of the rats were observed by HE staining. The levels of serum Hcy and matrix metalloproteinase-9 (MMP-9) were detected with ELISA,and the expression levels of CCR5 and CC chemokine ligand 5 (CCL5) proteins in thoracic aortic smooth muscle cells were detected by immunohistochemical staining. The expression levels of CCR5 and CCL5 mRNA in thoracic aortic smooth muscle cells were detected by RT-PCR. Results: The results of HE staining showed that the medial smooth muscle cells of the rat thoracic aorta were arranged neatly in control group, the intimal elastic membrane was complete and the endothelial cells were continuous; the smooth muscle cells of the thoracic aorta were loose in model group, the endometrium was damaged, the foam cells were formed under the intima, and the endothelial cells were necrotic;the pathological changes of the thoracic aorta of the rats in treatment group were between control group and model group. The levels of serum Hcy of the rats in model group and treatment group were higher than that in control group (P<0.05); the serum Hcy level in treatment group was lower than that in model group (P<0.05).The levels of serum MMP-9 of the rats in model group and treatment group were higher than that in control group (P<0.05);the serum MMP-9 level in treatment group was lower than that in model group (P<0.05).The expression gray values of CCR5 and CCL5 proteins in the thoracic aortic smooth muscle cells of the rats in rats in model group and treatment group were lower than those in control group (P<0.05); the expression gray values of CCR5 and CCL5 proteins in the thoracic aortic smooth muscle cells of the rats in treatment group were higher than those in model group (P<0.05).The relative expression levels of CCR5 and CCL5 mRNA in the thoracic aortic smooth muscle cells of the rats in model group and treatment group were higher than those in control group (P<0.05);the relative expression levels of CCR5 and CCL5 mRNA in the thoracic aortic smooth muscle cells of the rats in treatment group was lower than that in model group (P<0.05). Conclusion: Folic acid combined with vitamin B12 could decrease the serum Hcy and MMP-9 levels and the expression levels of CCR5 and CCL5 in the thoracic aortic smooth muscle cells which may be one of the mechanisms of folic acid combined with vitamin B12 in the treatment of atherosclerosis.

Key words: atherosclerosis, folic acid, vitamin B12, matrix metalloproteinase-9, CC-type chemokine receptor 5, CC-type chemotactic cytokine 5

中图分类号:

R363

徐吉光, 李宇宁. 叶酸联合维生素B12对高同型半胱氨酸血症引起大鼠动脉粥样硬化的治疗作用[J]. 吉林大学学报(医学版), 2017, 43(05): 943-947.

XU Jiguang, LI Yuning. Therapeutic effect of folic acid combined with vitamin B12 on atherosclerosis induced by hyperhomocysteinemia in rats[J]. Journal of Jilin University Medicine Edition, 2017, 43(05): 943-947.

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