妊娠相关血清蛋白-A对内皮细胞功能的影响及氟伐他汀的干预研究

发布时间：2018-05-04 00:03

本文选题：妊娠相关血清蛋白-A + 内皮细胞　；参考：《中南大学》2008年硕士论文

【摘要】： 目的观察妊娠相关血清蛋白-A(PAPP-A)在体外对内皮细胞功能的影响,同时探讨氟伐他汀对PAPP-A诱导内皮细胞功能改变的干预作用。方法体外培养人脐静脉内皮细胞株(HUVEC),将对数生长期的HUVEC随机分为四组。分别给予不同浓度的PAPP-A(0 ng/ml、50ng/ml、100 ng/ml、200 ng/ml)刺激,于0h、12 h、24 h和48 h收集上清液和细胞,用硝酸还原酶法测定一氧化氮(NO)的浓度,用免疫组化法测定内皮素-1(ET-1)的表达。取对数生长期的HUVEC随机分为五组,实验组分别予不同浓度的氟伐他汀(0 mmol/L、10~(-7)mmol/L、10~(-6)mmol/L、10~(-5)mmol/L)干预20min后,加入200 ng/mlPAPP-A,12h后收集上清液和细胞,用硝酸还原酶法测定NO的浓度,用免疫组化法测定ET-1的表达。结果在50ng/mlPAPP-A、100ng/mlPAPP-A及200ng/mlPAPP-A干预组,12h、24h和48h上清液NO浓度均低于同组0h NO的浓度,差异具有统计学意义,P＜0.05。在12h、24h和48h,随着PAPP-A浓度的增加,上清液NO的浓度逐渐降低,与对照组相比具有统计学意义,P＜0.05。在200ng/mlPAPP-A干预组,细胞ET-1的表达呈时间依赖性增加;在12h、24h和48h,随着PAPP-A浓度的增加,ET-1的表达呈浓度依赖性增加,与对照组相比具有统计学意义,P＜0.05。实验第二步,随着氟伐他汀干预浓度增加,上清液NO的浓度逐渐升高,与单纯PAPP-A干预组比较具有统计学意义,P＜0.05。在单纯PAPP-A干预组,细胞ET-1的表达明显增加,与对照组比较具有统计学差异,P＜0.05。随着氟伐他汀干预浓度增加,ET-1的表达逐渐减低,均低于单纯PAPP-A干预组,P＜0.05,但仍高于正常对照组,P＜0.05。结论PAPP-A在体外能诱导内皮细胞NO分泌减少,ET-1表达上调,且具有浓度依赖性,无明显时间依赖性。氟伐他汀对PAPP-A诱导的内皮细胞功能损害可能有保护作用,且具有浓度依赖性。
[Abstract]:Objective to observe the effect of pregnancy-associated serum protein (APP-A) on the function of endothelial cells in vitro, and to explore the effect of fluvastatin on the changes of endothelial cell function induced by PAPP-A. Methods Human umbilical vein endothelial cells (HUVECs) were cultured in vitro. HUVEC in logarithmic phase was randomly divided into four groups. The supernatants and cells were collected at 0 h ~ 12 h for 24 h and 48 h respectively. The concentration of nitric oxide (no) was determined by nitrate reductase method. The expression of endothelin-1 ET-1 was detected by immunohistochemical method. HUVEC in logarithmic growth phase was randomly divided into five groups. The experimental group was treated with different concentrations of fluvastatin 0 mmol / L 10 ~ 7 mmol / L ~ (10) ~ (-1) ~ (-1) ~ (-1) ~ (-1) / L ~ (10) mol / L ~ (10) ~ (-1) 20min, then added 200ng / ml PAPP-A for 12 h to collect supernatant and cell, and then the concentration of no was determined by nitrate reductase method, and the concentration of no was determined by nitric acid reductase method, and the concentration of no was determined by nitrate reductase method. The expression of ET-1 was detected by immunohistochemistry. Results at 50 ng / ml PAPP-A 100 ng / ml PAPP-A and 200ng/mlPAPP-A intervention group, the concentration of no in supernatant of 12h and 48h was lower than that of the same group (P < 0.05). At 12h and 48h, the concentration of no in supernatant gradually decreased with the increase of PAPP-A concentration, which was statistically significant compared with the control group (P < 0.05). In the 200ng/mlPAPP-A intervention group, the expression of ET-1 increased in a time-dependent manner, and increased in a concentration-dependent manner with the increase of PAPP-A concentration at 12h and 48h, which was significantly higher than that in the control group (P < 0.05). In the second step of the experiment, with the increase of fluvastatin concentration, the concentration of no in supernatant gradually increased, which was statistically significant compared with that of PAPP-A group (P < 0.05). The expression of ET-1 in PAPP-A group was significantly higher than that in control group (P < 0.05). With the increase of fluvastatin concentration, the expression of Tet-1 decreased gradually, which was lower than that of PAPP-A group (P < 0. 05), but still higher than that of normal control group (P < 0. 05). Conclusion in vitro, PAPP-A can induce no secretion in endothelial cells to decrease the expression of Tet-1 in a dose-dependent manner, but not in time dependent manner. Fluvastatin may have a protective effect on PAPP-A induced endothelial cell dysfunction, and it is in a concentration dependent manner.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363

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