AIM: To investigate the effects of atorvastatin on the expression of pregnancy - associated plasma protein A( PAPP - A )induced by TNF - α and IL - 1β in endothelial cells.METHODS: The rat aortic endothelial cells were isolated from thoracic aortas and cultured by the tissue explant method.The cells in passage 3-4 were used in the experiment and were randomly divided into 4 groups: blank control group: the cells were treated without any intervention; atorvastatin concentration groups: the cells were incubated with atorvastatin at the concentrations of 0.1, 1 and 10 μmol/L for 24 h; atorvastatin time groups: the cells were incubated with atorvastatin at the concentration of 10 μmol/L for 6 h,12 h and 24 h; atorvastatin + inflammatory factors groups: the cells were pre - incubated with 60 μg/L TNF - α or 20 μg/L IL -1β for 1 h, then different concentrations of atorvastatin ( 0.1, 1.0, 10 μmol/L ) were added for 6 h, 12 h and 24 h.MTT reduction assay was used to observe the cell proliferation.The mRNA expression of PAPP - A was detected by RT -PCR.The protein level of PAPP - A in the supernatants of cultured cells was measured by ELISA.RESULTS: Compared with blank control group, no significant change of cell proliferation was observed after the intervention of atorvastatin and TNF - α/IL - 1(3 for 3 h, 6 h, 12 h, 24 h and 48 h, indicating that the drugs had no toxic effects on the cells.No significant difference of PAPP - A expression between atorvastatin groups and blank control groups was found.Compared with TNF - α groups and IL - 1β groups, PAPP - A expressions in atorvastatin intervention groups significantly decreased.The protein level of PAPP - A was gradually decreased with the raised concentration of atorvastatin and the prolonged time in a concentration - and time - dependent manner.CONCLUSION: Atorvastatin doesn't influence the PAPP - A expression, but inhibits the expression of PAPP - A activated by inflammatory factors in a concentration - and time - dependent manner in primary cultured rat aortic endothelial cells.%目的:研究阿托伐他汀对炎症因子肿瘤坏死因子 α(TNF-α)和白细胞介素-1β(IL-1β)诱导大鼠主动脉内皮细胞表达妊娠相关血浆蛋白A(PAPP-A)的影响.方法:原代培养大鼠主动脉内皮细胞,选择生长良好的第3-4代细胞用于实验.实验分组:(1)空白对照组:不加干预措施原培养液继续培养;(2)阿托伐他汀浓度组:加入浓度分别为 0.1、1、10 μmol/L的阿托伐他汀作用24 h;(3)阿托伐他汀时间组:加入浓度为10 μmol/L的阿托伐他汀分别作用6、12、24 h;(4)阿托伐他汀干预组:加入TNF-α(60 μg/L)或者IL-1β(20 μg/L)作用1 h后,再加入不同浓度的阿托伐他汀(0.1、1、10 μmol/L)进行培养,分别于作用6、12、24 h终止实验.MTT法观察药物对细胞增殖的影响,RT-PCR技术测定细胞中PAPP-A mRNA的表达,ELISA方法检测上清液中PAPP-A蛋白水平.结果:(1) 外源性加入阿托伐他汀及TNF-α或者IL-1β后,分别作用3、6、12、24、48 h,同对照组相比,细胞增殖能力无明显变化(P<0.05),说明干预药物本身对细胞没有毒性作用.(2)单独加入阿托伐他汀,细胞PAPP-A表达水平与空白对照组比较没有明显差异(P<0.05).(3)炎症因子作用一段时间后再加入阿托伐他汀,PAPP-A mRNA和蛋白表达水平随着阿托伐他汀浓度和作用时间的增加而逐渐降低,有一定的浓度-时间依赖关系,与 TNF-α组和IL-1β组比较明显下降(P<0.05).结论:阿托伐他汀本身对细胞表达PAPP-A没有影响,但在一定程度上抑制TNF-α和IL-1β诱导的内皮细胞PAPP-A的表达,并有浓度-时间依赖关系.
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