铅、镉联合对SD大鼠肾细胞的毒性损伤及硒的保护作用

发布时间：2018-08-07 12:19

【摘要】： 铅(Pb)、镉(Cd)均为重要的环境污染物,并通过食物链系统进入动物和人体产生毒性作用。本研究是在建立SD大鼠肾细胞培养的基础上,通过培养液中加入不同浓度的Cd(10、20、40μmol/L)、Pb(10,20,40μmol/L)和Pb-Cd联合(分别为5、10、20μmol/L),并加入Na_2SeO_30.5mg/L,在培养6h后,观察细胞形态、测定细胞凋亡率和细胞周期及上清液脂质过氧化指标,探讨Pb、Cd对肾细胞的联合毒性及硒的保护作用。结果表明: (1)建立了0.05%胰蛋白酶消化SD乳鼠肾组织培养肾细胞的方法,该法细胞产量高,操作简便,不易污染,花费较少。 (2)低浓度Pb或Cd(10μmol/L)对细胞无明显损伤,20μmol/L以上可以使细胞皱缩、体积变小,表面有细胞碎片,甚至呈泡沫状或树枝状。Pb、Cd联合可加剧细胞损伤。 (3)低浓度的Pb、Cd(10μmol/L)使细胞凋亡率下降,而20μmol/L以上的Pb、Cd均可使细胞凋亡率增加,并改变细胞周期。Pb、Cd联合具有明显的协同性,分别浓度达40μmol/L时,细胞凋亡率降低,表明细胞已有死亡。 (4)Pb、Cd均可引起肾细胞培养液GSH-Px、SOD的活性降低,MDA含量升高,并存在剂量和时间效应。 (5)在培养液中添加Na_2SeO_3,可降低肾细胞凋亡率,使细胞周期趋于正常,并缓解GSH-Px、SOD活性下降和MDA含量升高的幅度,表明Se对Pb、Cd所致的肾细胞损伤具有一定的保护作用。
[Abstract]:Lead, (Pb), cadmium and (Cd) are important environmental pollutants and have toxic effects on animals and human beings through food chain system. Based on the establishment of renal cell culture in SD rats, different concentrations of CD (10 ~ 20 ~ 40 渭 mol/L) Pb (10 ~ 20 ~ 20 ~ 40 渭 mol/L) and Pb-Cd (5 ~ 10 ~ 10 ~ 20 渭 mol/L) were added to the culture medium to observe the morphology of the cells, and Na2SeOO _ (30) mg / L was added to the culture medium. The apoptosis rate, cell cycle and lipid peroxidation in supernatant were measured to study the combined toxicity of PBN and CD on renal cells and the protective effect of selenium. The results showed that: (1) A method for digestion of kidney cells by 0.05% trypsin was established. The method has the advantages of high cell yield, simple operation and easy contamination. (2) low concentration of Pb or CD (10 渭 mol/L) had no obvious damage to the cells. More than 20 渭 mol/L could make the cells shrink, become smaller, and have cell fragments on the surface. (3) low concentration of PBN CD (10 渭 mol/L) decreased the apoptosis rate of cells, but more than 20 渭 mol/L of PBN CD could increase the apoptosis rate of the cells, and even the combination of foam or dendritic cells could aggravate the cell injury. (3) low concentration of PBN CD (10 渭 mol/L) decreased the apoptosis rate of the cells, but more than 20 渭 mol/L, the apoptosis rate of the cells increased. When the cell cycle was changed, the cell apoptosis rate decreased when the concentration of 40 渭 mol/L was 40 渭 mol/L, respectively. The results indicated that the cells had died. (4) PbPxC could induce the decrease of the activity of GSH-PxN SOD in renal cell culture medium and the increase of MDA content. There were dose and time effects. (5) adding NaCo2SeO3 to the culture medium could reduce the apoptosis rate of renal cells and make the cell cycle normal. It also alleviated the decrease of SOD activity and the increase of MDA content, indicating that se has a certain protective effect on renal cell injury induced by PBH and CD.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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