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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、400μmol/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;Ed U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-αmRNA及蛋白表达水平。结果(1)FFA在400μmol/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+HMWHA组、FFA+LMW-HA组IL-6、IL-8、TNF-αmRNA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-αmRNA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。
Objective To investigate the effect of hyaluronic acid (HA) on the expression of interleukin 6 (IL-6), interleukin 8 and tumor necrosis factor α (TNF-α) in human vascular smooth muscle cells (HA- VSMCs) induced by free fatty acids )Impact. Methods HA-VSMCs were cultured and treated with different concentrations of FFA and different concentrations of FFA + HMW-HA or LMW-HA for 24,48 h. The effect of FFA and FFA + HA on the survival rate of HA-VSMC was detected by thiazolyl blue assay. The cell proliferation activity was detected by Ed U staining. The contents of IL-6, IL-8 and TNF-α in culture supernatant were detected by enzyme-linked immunosorbent assay The mRNA and protein levels of IL-6, IL-8 and TNF-α were detected by quantitative PCR and Western blot. Results (1) FFA induces the proliferation of HA-VSMC in a concentration-dependent manner at a concentration of 400 μmol / L. (2) Both HMW-HA and LMW-HA inhibited the proliferation of HA-VSMC induced by FFA and the inhibitory effect of HMW-HA In LMW-HA; (3) FFA significantly promoted the secretion of IL-6, IL-8 and TNF-α in HA-VSMC with significant time and concentration dependent; HA inhibited the above effects; (4) PCR and Western blot results showed that the levels of IL-6, IL-8 and TNF-αmRNA and protein in FFA + HMWHA group and FFA + LMW-HA group were lower than those in FFA group (all P <0.01) HA group was lower than that of FFA + LMW-HA group (P <0.01). CONCLUSION: (1) The expression of IL-6, IL-8 and TNF-α mRNA and protein in HA-VSMC induced by FFA is increased. (2) The expression of IL-6, IL-8 and TNF- Inhibited, and HMW-HA inhibition stronger than LMW-HA.