几种因子对HaCaT细胞合成结合珠蛋白影响的研究

发布时间：2018-11-13 20:09

【摘要】：目的 在基因及蛋白质水平上研究白细胞介素-6,白细胞介素-4对人角质形成细胞:HaCaT细胞合成结合珠蛋白的影响。 探讨肿瘤坏死因子α及干扰素γ对HaCaT细胞合成结合珠蛋白的影响。 探讨免疫抑制剂地塞米松及甲氨喋呤对HaCaT细胞合成结合珠蛋白的影响。 材料和方法 1.细胞培养 将HaCaT细胞接种于六孔板内,在含有10%胎牛血清(fetal calf serum,FCS),1mM丙酮酸盐,2mM谷氨酰胺,青霉素(100μg/mL),链霉素(100μg/mL)的1640培养液中,5% CO_2,37℃条件下培养。细胞形成单层后加入含有不同刺激因子的,含有或不含1μM地塞米松的,不含有胎牛血清的1640培养液。所用刺激因子如下:白细胞介素-6(interleukin-6,50ng/mL),白细胞介素-4(interleukin-4,20ng/mL),肿瘤坏死因子α(tumor necrosis factor α,20ng/mL),干扰素γ(interferon γ,20ng/ml),地塞米松(dexamethasone,10μM,100μM,1000μM)及甲氨喋呤(methotrexate,50ng/ml,500ng/ml,5000ng/ml)。分别作用12,24,48小时后,收集细胞及细胞上清,用于结合珠蛋白(Haptoglobin,Hp)的检测。 2.实验方法 (1) 半定量RT-PCR ① 细胞总RNA的提取:Trizol RNA提取试剂,由3×10~6细胞中提取总RNA。
[Abstract]:Objective to study the effects of interleukin-6 and interleukin-4 on the synthesis of globin in human keratinocytes: HaCaT cells at the gene and protein levels. To investigate the effect of tumor necrosis factor 伪 and interferon 纬 on the synthesis of globin in HaCaT cells. To investigate the effects of immunosuppressants dexamethasone and methotrexate on the synthesis of globin in HaCaT cells. Materials and methods 1. HaCaT cells were inoculated into six well plates in cell culture. The cells containing 10% fetal bovine serum (fetal calf serum,FCS, 1mM pyruvate, 2mM glutamine, penicillin (100 渭 g/mL) were cultured in vitro. Streptomycin (100 渭 g/mL) was cultured in 1640 medium at 5% CO_2,37 鈩，

本文编号：2330296