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透明质酸对游离脂肪酸引起的人主动脉血管平滑肌细胞表达IL-6、IL-8和TNF-α的影响
作者:
作者单位:

(1.宁夏医科大学临床医学院检验系,宁夏银川市 750003;2.甘肃省人民医院中心实验室,甘肃省兰州市 730000 ;3.甘肃省人民医院内分泌科 甘肃省内分泌代谢病重点实验室,甘肃省兰州市 730000;4.临床检验中心,甘肃省兰州市 730000)

作者简介:

杨兴文,硕士研究生,检验医师,主要从事血管疾病的生物化学及分子生物学研究,E-mail为yangxingwen199051@163.com。

基金项目:

国家自然科学基金资助项目(81460155、81260136、30960147)


Effect of hyaluronic acid on the expressions of IL-6, IL-8 and TNF-α in human aortic vascular smooth muscle cells induced by free fatty acid
Author:
Affiliation:

1.Inspection Department, Clinical Medical College, Ningxia Medical University, Yinchuan, Ningxia 750003, China;2.Central Laboratory, ;3.Department of Endocrinology & Key Laboratory of Endocrine and Metabolic Diseases of Gansu Province, ;4.Clinical Laboratory Center, People's Hospital of Gansu Province, Lanzhou, Gansu 730000, China)

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    摘要:

    目的 探讨透明质酸(HA)对游离脂肪酸(FFA)引起的人主动脉血管平滑肌细胞(HA-VSMC)表达白细胞介素6(IL-6)、IL-8和肿瘤坏死因子α(TNF-α)的影响。方法 培养HA-VSMC,用不同浓度(0、100、200、400 μmol/L)FFA和不同浓度FFA＋高分子量HA(HMW-HA)或低分子量HA(LMW-HA)处理细胞24、48 h。噻唑蓝法检测FFA和FFA＋HA对HA-VSMC存活率的影响；EdU染色检测细胞增殖活性；酶联免疫吸附法检测培养上清IL-6、IL-8和TNF-α含量；实时荧光定量PCR和Western blot检测IL-6、IL-8和TNF-α mRNA及蛋白表达水平。结果 (1)FFA在400 μmol/L内呈浓度依赖性诱导HA-VSMC增殖；(2)HMW-HA、LMW-HA均对FFA诱导的HA-VSMC增殖有抑制作用,且HMW-HA抑制作用强于LMW-HA；(3)FFA显著促进HA-VSMC IL-6、IL-8和TNF-α的分泌,且有明显的时间和浓度依耐性；HA可以抑制上述作用；(4)实时荧光定量PCR和Western blot结果显示,FFA＋HMW-HA组、FFA＋LMW-HA组IL-6、IL-8、TNF-α mRNA及蛋白表达水平均低于FFA组(均P＜0.01),其中FFA＋HMW-HA组又低于FFA＋LMW-HA组(P＜0.01)。结论 (1)FFA诱导HA-VSMC IL-6、IL-8、TNF-α mRNA和蛋白表达升高；(2)HA对FFA引起的HA-VSMC IL-6、IL-8和TNF-α的表达有抑制作用,且HMW-HA抑制效果强于LMW-HA。

    Abstract:

    Aim To investigate the effect of hyaluronic acid (HA) on the expression of interleukin-6 (IL-6), IL-8 and tumor necrosis factor-α (TNF-α) in human aortic vascular smooth muscle cell (HA-VSMC) induced by free fatty acid (FFA). Methods HA-VSMCs were cultured and dealt with different concentrations (0,0, 0,0 μmol/L) of FFA and FFA plus high molecular weight HA (HMW-HA) or low molecular weight HA (LMW-HA) for 4,8 hours. The effects of FFA and FFA plus HA on the survival rate of HA-VSMCs were detected by methyl thiazolyl tetrazolium (MTT) method. Cell proliferation activity was detected by EdU staining. The contents of IL-6, IL-8 and TNF-α in culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Real-time fluorescence quantitative PCR and Western blot were used to detect the expression levels of IL-6, IL-8 and TNF-α mRNA and protein. Results(1)FFA showed a concentration-dependent induction to HA-VSMC proliferation in the 400 μmol/L. (2)HMW-HA and LMW-HA both inhibited the proliferation of HA-VSMC induced by FFA, and the inhibitory effect of HMW-HA was stronger than that of LMW-HA. (3)FFA significantly promoted the secretions of HA-VSMC IL-6, IL-8 and TNF-α in a apparent time- and concentration-dependent manner; HA could inhibit the above effect. (4)Real-time fluorescence quantitative PCR and Western blot results showed that the expression levels of IL-6, IL-8, TNF-α mRNA and protein in FFA＋HMW-HA group and FFA＋LMW-HA group were lower than those in FFA group (all P＜0.01), where the FFA＋HMW-HA group was lower than the FFA＋LMW-HA group (P＜0.01). Conclusions (1)FFA can induce the increases of IL-6, IL-8 and TNF-α mRNA and protein expressions in HA-VSMCs. (2)HA can inhibit the expressions of IL-6, IL-8 and TNF-α induced by FFA, and the inhibitory effect of HMW-HA is stronger than that of LMW-HA.

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杨兴文,李伟华,李元,董莉,权金星,韩平治.透明质酸对游离脂肪酸引起的人主动脉血管平滑肌细胞表达IL-6、IL-8和TNF-α的影响[J].中国动脉硬化杂志,2017,25(2):115~121.

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  • 收稿日期:2016-07-27
  • 最后修改日期:2016-08-31
  • 在线发布日期: 2017-02-08

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