中药含药血清对高糖诱导心肌细胞促炎因子巨噬细胞移动抑制因子表达的影响

Effect of herbal serum on macrophage migration inhibitory factor expression induced by high glucose in myocardial cells

目的探讨益气化痰清毒方煎剂对巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)水平的影响及其含药血清对高糖诱导心肌细胞株(H9C2)MIF表达的影响。方法制备中药煎剂(药物为黄芪、党参、毛冬青等),取40只SD大鼠随机(随机数字表法)分4组(空白对照组、低、中、高剂量组),完成灌药后用酶联免疫吸附试验(ELISA)法测各组动物血清MIF浓度;制备各组含药血清,分别干预经33 mmol/L葡萄糖处理后的H9C2心肌细胞,用反转录聚合酶链反应(RT-PCR)法测定各组细胞MIF mRNA的表达。结果与对照组相比,经低、中、高剂量药物干预的大鼠血清MIF浓度均呈减少趋势,差异有统计学意义(P<0.05)。经高糖处理的H9C2细胞MIF表达明显增加,在含药血清干预后MIF表达明显减少,且各组含药血清对MIF表达均有明显抑制作用,差异均有统计学意义(P<0.05),其中高剂量组效果最显著(P<0.01)。随着药物浓度的增加,MIF表达呈减少趋势。结论益气化痰清毒方可降低大鼠血清MIF浓度,其含药血清对高糖诱导H9C2心肌细胞MIF表达有明显抑制作用。

Objectives To investigate the effects of the Yiqihuatanqingdu decoction on serum concentrations of macrophage migration inhibitory factor （MIF） and how the herbal serum affect MIF mRNA expression induced by high glucose in H9C2 cells.Methods The herbal decoction which consisted of radix astragali,radix codonopsis,radix ilicis pubescentis and so on were prepared.According to the herbal decoction concentration,40 SD rats were randomly divided into four groups：control group,low,medium and high dose groups.Their serum MIF concentrations were determined by enzyme-linked immunosorbent assay （ELISA）.H9C2 cells cultured with 33 mmol/L of glucose were treated with three groups of herbal serum.MIF mRNA expressions were detected by reverse transcription polymerase chain reaction （RT-PCR）.Results Compared with control group,serum concentrations of MIF significantly decreased in low to high dose groups （P＜0.05）.Expression of MIF in H9C2 cells cultured with high glucose significantly increased,but significantly decreased after treated with herbal-containing serum.Expression of MIF in the three groups were significantly inhibited by herbal-containing serum （P＜0.05）,especially in high dose group （P＜0.01）.Herbal-containing serum decreased the expression of MIF mRNA in a dose-dependent manner.Conclusions The Yiqihuatanqingdu decoction can significantly inhibit the serum concentration of the pro-inflammatory factor MIF in rats and its containing serum can decrease the expression of MIF mRNA in H9C2 cells induced by high glucose.