苦参素对人肝正常细胞L02影响的实验研究

An experimental study of the effect of oxymatrine on human normal hepatocyte cell line L02

目的探讨不同浓度苦参素(OM)作用不同时间对人肝正常细胞L02的损伤作用或不良反应。方法体外培养人肝正常细胞L02;采用MTT法观察不同浓度的OM作用不同时间后观察肝正常细胞L02细胞形态变化及增殖的情况。结果倒置显微镜下观察:与阴性对照组相比较,OM作用时间在24h、浓度为0～2.0mg/ml时L02细胞的形态及数量无明显变化,当作用时间超过24h后,随着时间延长、浓度增加,OM实验组L02细胞收缩变小、与周围细胞分离、细胞核周围较多空泡样结构均越明显,细胞数量逐渐减少。MTT检测结果显示:①当作用时间在24h时,与阴性对照组相比较,OM浓度为0.5mg/ml、1.0mg/ml、2.0mg/ml时对正常肝细胞L02的增殖无明显抑制作用(P>0.05),浓度为3.0mg/ml、4.0mg/ml、8.0mg/ml时对人肝正常细胞增殖有不同程度的抑制作用(P<0.05),其抑制率分别为8.48%、15.43%、51.72%;②OM 1.0mg/ml、2.0mg/ml、3.0mg/ml、4.0mg/ml、8.0mg/ml各浓度组作用于L02细胞48h和72h的抑制率分别为18.79%、26.06%、27.58%、31.52%、76.36%和33.20%、38.53%、41.82%、55.61%、89.95%。与24h相比较,作用48h、72h对正常肝细胞的增殖有不同程度的抑制作用(P<0.05或P<0.01)。结论 OM在低浓度短时间内对人正常肝细胞L02无明显损伤或不良反应,但随着浓度增加、作用时间延长对人正常肝细胞L02的增殖有不同程度的损伤或不良反应,且呈时间-剂量依赖性。

Objective To explore liver cell L02 at different time points. the effect of different concentrations Methods Human normal liver cells of oxymatrine on human normal L--02 were cultured in vitro and treated with oxymatrine. After cells were exposed to different concentrations of oxymatrine at different time points, then observed the morphological changes in was performed to detect the L02 cell proliferation. scope, compared with the negative control group, L02 cells by using inverted microscope, and MTT assay Results The cells were observed under inverted micro- when the cells exposed to oxymatrine at 24h and at 0 2.0mg/mlconcentration, L02 cell morphology and number did not change significantly, while after 24h, with the prolongation of time and increase of concentration, we found that in the experimental group, L02 cells shrank and separated with each other, what＇s more, we can also found around the nuclei more and more vacuole --like structures formed, the number of cells were less and less. The MTT assay results showed that.@ When the cells were exposed to oxymatrine at 24h, compared with the negative control group, there was no prolifera- tion inhibition at 0.5mg/ml, 1.0mg/ml, 2.0mg/ml concentration of oxymatrine in experimental groups （ P 0.05）, when the oxymatrine concentration increased to 3.0mg/ml, 4.0mg/ml, 8.0mg/ml, there were differ- ent degrees of inhibition for L02 cells proliferation （ P〈 0.05）, the inhibition rates were 8.48%, 15.43%, 51.72%. （2） The inhibition rate of oxymatrine at the concentration of 1. 0mg/ml, 2. 0mg/ml, 3. 0mg/ml, 4. Omg/mland 8. 0mg/ml on human normal liver cells L-02 for 48h and 72h were 18.79%, 26. 06%, 27.58%, 31.52%, 76.36% and 33. 20%, 38. 53%, 41. 82%, 55. 61%, 89. 95%, respectively. Compared with 24h, ceils exposed to oxymatrine at 48h and 72h could inhibit on normal liver cell proliferation at different degrees （ P 〈0.05 or P 〈0. 01 ）. Conclusion Oxymatrine at low concentrations has no significant injury or adverse effects on human normal cells 1.02 within a short period of time, but with the increase of oxymatrine concentra- tion and tile exposure time prolongation, there have different degrees of injury or side effects on human normal liver ceils [.02. and has time-- and dose-- dependence.