摘要
目的观察普罗布考对人视网膜色素上皮(Human retinal pigment epithelial,hRPE)细胞抗过氧化氢(Hydrogen peroxide,H_2O_2)损伤的影响。方法以传代培养hRPE细胞为研究对象,预处理组根据普罗布考浓度不同分成4个亚组(20μmol/L组、40μmol/L组、60μmol/L组、80μmol/L组),各亚组普罗布考作用24h之后加入500μmol/L H_2O_2作用24h建立氧化损伤模型;单纯损伤组:损伤及培养条件同预处理组,但不加普罗布考;正常对照组:相同条件培养,不加普罗布考且不用H_2O_2损伤。应用比色法测定乳酸脱氢酶(Lactate dehydrogenase,LDH)细胞释放率和细胞脂质过氧化产物丙二醛(Maleic dialdehyde,MDA)含量;免疫细胞化学方法检测细胞内核因子Kappa B(Nuclear factor kappa B NF-κB)的表达情况。结果 DH释放率、MDA含量及NF-κB核移位率,单纯损伤组较正常对照组明显升高,各预处理组与单纯损伤组相比均明显降低,预处理组之间随着普罗布考的浓度增加,LDH释放率、MDA含量以及NF-κB核移位率均逐渐降低(P<0.05)。线性相关分析结果显示,普罗布考浓度与LDH细胞释放率、MDA含量、NF-κB核移位率均呈负相关。正常对照组细胞中NF-κB免疫细胞化学染色呈阴性表达。单纯损伤组内几乎所有细胞NF-κB均呈核阳性染色。预处理组随着普罗布考浓度增大,NF-κB细胞核阳性染色数逐渐减少,染色程度亦逐渐减低。结论普罗布考预处理对hRPE细胞的氧化损伤具有一定的保护作用,其机制可能与抑制NF-κB的活化有关。
Objective To investigate the protective effects of Prohucol on human retinal pigment epithelial cells against oxidative damage induced by hydrogen peroxide. Methods The subeultured hRPE cells was as study target. Pretreatment group were pretreated 24 hours with different concentrations （ 20μmol/L, 40μmol/L,60μmol/L and 80μmol/L） respectively of Prohucol solution. They were treated with 500μmol/L of H2O2 for 24 hours to establish the cell injury model. Simply injury group without probucol; normal control group without Probucol and H2O2. The content of Malondialdehyde（MDA） which was the product of cellular lipid peroxidation and the releasing rate of lactate dehydrogenase（LDHI were estimated by chroma- tometry. NF-κ3 was measured by immunohistochemistry. Results The release rate of LDH,the level of MDA,the nuclear translocation rate of NF-κ3 in simply injury group were significant higher than control group, whereas the level in pretreatment group were lower than simply injury group. Meanwhile, the pa- rameter in different concentrations of Probucol treatment groups were decreased accompanied with the increase of concentration. Conclusion Probucol can improve the antioxidative stress status of hRPE cells. The possible mechanism may attribute to the protective effect in inhibiting the activation of NF -κB in hRPE cells.
出处
《中国煤炭工业医学杂志》
2016年第5期718-722,共5页
Chinese Journal of Coal Industry Medicine
关键词
普罗布考
视网膜色素上皮细胞
氧化损伤
NF-ΚB
Probucol
Retinal pigment epithelial cells
Oxidative damage
Cell culture techniques
NF-κB
