摘要
目的探讨橙皮素(HES)对转化生长因子-β_1(TGF-β_1)诱导心肌成纤维细胞增殖的影响,并探讨其可能的机制。方法取SD大鼠30只,取其心肌组织进行成纤维细胞的培养。而后采用TGF-β_1(10 ng/ml)刺激成纤维细胞,采用4种不同浓度梯度的HES(25μmol/L、50μmol/L、75μmol/L、100μmol/L)干预成纤维细胞24 h,以台盼蓝染色检测其对细胞活性的影响;荧光酶标仪检测法用于检测细胞内活性氧(ROS)的水平;活细胞计数试剂盒(CCK-8)用以检测成纤维细胞的增殖情况。结果台盼蓝染色结果提示,HES对成纤维细胞的细胞活性无显著影响,各组活性值分别为空白对照组:100%,HES 25μmol/L组:95%,HES 50μmol/L组:94%,HES 75μmol/L组:93%,HES100μmol/L组:93%。酶标仪检测结果表明,与空白对照组相比,TGF-β_1可增加成纤维细胞内ROS的水平(P<0.01),当HES与TGF-β_1同时作用时,ROS水平随着HES浓度的增加逐渐降低,均存在统计学意义(P<0.05);TGF-β_1可刺激成纤维细胞增殖(P<0.01),当HES(100μmol/L)或N-乙酰半胱氨酸(NAC)(1 mmol/L)与TGF-β_1同时作用时,可明显抑制成纤维细胞增殖(P<0.01)。结论 HES可通过抑制TGF-β_1诱导的氧化应激进而抑制心肌成纤维细胞增殖,提示其对于预防心肌纤维化可能具有潜在效果。
Objective To investigate the effect of hesperetin（HES） on transforming growth factor-β1（TGF-β1） cardiac fibroblasts induced cell proliferation,and to explore its possible mechanism.Methods 30 SD rats were included;take the myocardial tissues to culture the fibroblasts.And then uses the TGF-β1（10 ng/ml） to stimulate fibroblast cells,with 4different concentrations of HES（25 μmol/L,50 p,mol/L,75 μmol/E,100 μmol/L） to intervene fibroblasts for 24 h,blue staining was used to detect the effect on cell viability by trypan;enzyme-linked fluorescent immunoassay testing method was used for the detection of intracellular reactive oxygen species（ROS） level;living cell counting Kit（CCK-8） was used to detect the proliferation of fibroblasts.Results Trypan blue staining results indicated that,HES had no significant effect on fibroblast activity in cells,each activity values were blank control group：100%,HES 25 μmol/1.group：95%,HES 50 μmol/L group;94%,HES 75 μmol/L group：93%,HES 100 μmol/1.group：93%,enzyme mark instrument resulls show that,compared with the blank control group,TGF-β1 can increase the level of ROS in fibroblast cells（P〈0.01）,when HES and TGF-β1 have effect at the same lime,ROS levels decreased gradually with the increase of HES concentration,there were statistical significance（P〈0.05）;TGF-β1 could stimulate the proliferation of fibroblasts（P〈0.01）,when HES（100μmol/L） or N-acetylcysleine（NAC）（1 mmol/L） and TGF-β1 have effect at the same lime,they can inhibit fibroblast proliferation（P〈0.01）.Conclusion Through inhibiting TGF-β1 induced oxidative stress,HES can inhibit the proliferation of cardiac fibroblasts,which suggested that it may have potential for the prevention of myocardial fibrosis.
出处
《疑难病杂志》
CAS
2015年第4期376-379,共4页
Chinese Journal of Difficult and Complicated Cases
基金
中央高校基本科研业务费专项基金资助项目(No.2012302020212)
