5-甲基四氢叶酸对大鼠动脉粥样硬化的潜在干预作用

doi:10.6040/j.issn.1671-7554.0.2021.1598

摘要/Abstract

摘要： 目的探讨5-甲基四氢叶酸(5-MTHF)对大鼠动脉粥样硬化的潜在干预特性和可能机制。方法高脂饮食联合维生素D2建立大鼠动脉粥样硬化模型,将48只雄性Wister大鼠随机分为对照组、模型组、5-MTHF低剂量组(0.5 mg/kg)和5-MTHF高剂量组(2 mg/kg),每组12只。6周后处死大鼠,检测各组大鼠血清活性叶酸、同型半胱氨酸、血脂、氧化应激、炎症因子和内皮因子含量,采用苏木精-伊红染色法观察大鼠主动脉病理变化,采用qRT-PCR法检测大鼠主动脉中核因子-κB p65(NF-κB p65)和凝集素样氧化型低密度脂蛋白受体-1(LOX-1)mRNA表达。结果与对照组比较,模型组大鼠体质量下降(P<0.05),血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDC-C)含量增加(P<0.001),高密度脂蛋白(HDL-C)含量降低(P<0.001),主动脉中有平滑肌细胞增生和脂质斑块弥漫,提示大鼠动脉粥样硬化造模成功。模型组血清活性叶酸含量较对照组差异无统计学意义(P>0.05),同型半胱氨酸浓度增加77.47%(P<0.001)。与模型组相比,5-MTHF低剂量组、5-MTHF高剂量组血清活性叶酸水平升高(P=0.042,P<0.001),同型半胱氨酸浓度降低(P=0.046,P<0.001);5-MTHF低剂量组大鼠的血脂异常、一氧化氮(NO)和一氧化氮合酶(NOS)活力、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、炎症肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)、白介素6(IL-6)、内皮素1(ET-1)、可溶性细胞间黏附分子1(sICAM-1)、NF-κB p65和LOX-1 mRNA表达水平均无明显改善(P>0.05);5-MTHF高剂量组大鼠血清TC和LDC-C含量降低(P=0.023,P=0.036),HDL-C含量升高(P=0.035);NO浓度、NOS和SOD活力增加(P=0.035,P=0.022,P=0.04);MDA、TNF-α、IL-6、ET-1含量降低(P=0.022,P=0.045,P=0.024,P=0.045),GSH-Px活力明显增强(P=0.007);LOX-1 mRNA表达下调(P=0.038),NF-κB p65 mRNA表达下调差异无统计学意义(P>0.05)。5-MTHF对动脉粥样硬化大鼠斑块无消退作用,但高剂量5-MTHF可减少平滑肌细胞增生和斑块中泡沫细胞聚集。结论 5-MTHF可以改善动脉粥样硬化大鼠的血脂异常、氧化应激和炎症反应,减少泡沫细胞聚集和平滑肌细胞增生,但未减轻动脉粥样硬化斑块,其作用机制可能是降低同型半胱氨酸水平,抑制氧化应激和下调LOX-1 mRNA表达。

关键词: 5-甲基四氢叶酸, 动脉粥样硬化, 同型半胱氨酸, 氧化应激, 凝集素样氧化型低密度脂蛋白受体-1

Abstract: Objective To investigate the potential intervention properties and possible mechanism of 5-methyltetrahydrofolate(5-MTHF)on atherosclerosis in rats. Methods Rat atherosclerosis models were established with a high-fat diet combined with vitamin D2. A total of 48 male Wister rats were randomly divided into control group, model group, 5-MTHF low-dose and high-dose groups(0.5 mg/kg, 2 mg/kg), with 12 rats in each group. The rats were sacrificed 6 weeks later, and the contents of folate, homocysteine, blood lipid, oxidative stress, inflammatory factors, and endothelial factors in the serum were measured. The pathological changes in the aorta were observed with HE staining. The mRNA expressions of nuclear factor-κB p65(NF-κB p65)and lectin-like oxidized low-density lipoprotein receptor-1(LOX-1)in aorta were detected with qRT-PCR. Results Compared with the control group, the model group had decreased body weight(P<0.05), increased levels of serum total cholesterol(TC), triglyceride(TG), low-density lipoprotein(LDC-C)(P<0.001), decreased level of high-density lipoprotein(HDL-C)(P<0.001), and smooth muscle cell hyperplasia and diffuse lipid plaques in the aorta, suggesting that the atherosclerosis models were established successfully. The serum folate content in the model group was not significantly different from that in the control group(P>0.05), but the homocysteine concentration increased by 77.47%(P<0.001). Compared with the model group, the 5-MTHF low- and high-dose groups had increased serum folate level(P=0.042, P<0.001)but decreased homocysteine concentration(P=0.046, P<0.001). Low-dose 5-MTHF did not significantly improve dyslipidemia, nitric oxide synthase(NOS), superoxide dismutase(SOD), glutathione peroxidase(GSH-Px)activities, nitric oxide(NO), malondialdehyde(MDA), inflammatory tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6), endothelin-1(ET-1), soluble intercellular adhesion molecule 1(sICAM-1), and mRNA expressions of NF-κB p65 and LOX-1(P>0.05). In the 5-MTHF high-dose group, serum TC and LDC-C contents decreased(P=0.023, P=0.036), HDL-C level increased(P=0.035); NO concentration, NOS and SOD activities increased(P=0.035, P=0.022, P=0.04); MDA, TNF-α, IL-6 and ET-1 contents decreased(P=0.022, P=0.045, P=0.024, P=0.045), but the GSH-Px activity was significantly enhanced(P=0.007); the mRNA expression of LOX-1 was down-regulated(P=0.038)but the down-regulation of the mRNA expression of NF-κB p65 was not statistically significant(P>0.05). 5-MTHF had no regression effects on plaques in atherosclerotic rats, but high-dose 5-MTHF reduced smooth muscle cell proliferation and foam cell aggregation in plaques. Conclusion 5-MTHF can improve dyslipidemia, oxidative stress and inflammation, reduce foam cell aggregation and smooth muscle cell proliferation, but does not reduce atherosclerotic plaques in atherosclerotic rats, probably by reducing homocysteine level, inhibiting oxidative stress and down-regulating the mRNA expression of LOX-1.

Key words: 5-methyltetrahydrofolate, Atherosclerosis, Homocysteine, Oxidative stress, Lectin-like oxidized low-density lipoprotein receptor-1

中图分类号:

R543.5

吴虹,张正铎,唐延金,祁少俊,高希宝. 5-甲基四氢叶酸对大鼠动脉粥样硬化的潜在干预作用[J]. 山东大学学报 (医学版), 2022, 60(8): 6-13.

WU Hong, ZHANG Zhengduo, TANG Yanjin, QI Shaojun, GAO Xibao. Potential intervention effects of 5-methyltetrahydrofolate on atherosclerosis in rats[J]. Journal of Shandong University (Health Sciences), 2022, 60(8): 6-13.

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