摘要
目的建立SD大鼠神经星形胶质细胞对不同浓度一氧化氮诱导产生Hif-1α模型,观察不同浓度一氧化氮对胶质细胞缺氧诱导因子的调控及对细胞的损伤。方法使用不同浓度(0.3-1.2μmol/L)DETA诱导大鼠神经胶质细胞,蛋白印迹法检测HIF-1α及其下游编码蛋白VEGF蛋白水平和mRNA表达,同时采用CCK-8法、化学比色法、TUNEL法测定在不同一氧化氮浓度下对细胞的增殖、氧化应激反应和细胞凋亡水平的影响。结果HIF-1α及其下游编码蛋白VEGF蛋白水平和mRNA表达随DETA浓度的增加而升高,差异有统计学意义(P<0.01),当DETA达到1.2 mmol/L浓度时并未引起HIF-1α和VEGF蛋白水平及mRNA的进一步升高;随着DETA预处理浓度(0.3-1.2mmol/L)不断增高,对照阴性组结果,细胞增殖降低,MDA水平明显增高(P<0.05),细胞凋亡率明显增加(P<0.01)。结论推荐0.9mmol/L浓度DETA预处理胶质细胞12 h后,对细胞损伤较轻,HIF-1α蛋白水平及mRNA的表达相对较高,是建立SD大鼠神经星形胶质细胞外源性一氧化氮诱导产生Hif-1α模型理想浓度。
Objective To establish a model of H if-1 production induced by neuroastrocytes of SD rats,and to observe the regulation of glial hypoxia-inducing factors at different concentrations.Methods DETA was induced by different concentrations(0.3-1.2μmol/L)by protein blotting,and the proliferation,oxidative stress response and apoptosis were measured by CCK-8,chemical color and TUNEL methods.Result HIF-1 alpha and downstream encoding protein expression level of VEGF protein and mRNA increased with the increase of the concentration of DETA,the difference was statistically significant(P<0.01).When D ETA reached the concentration of1.2 m mol/L,it did not cause a further increase in HIF-1 and VEGF protein levels and mRNA;With the increasing pretreatment concentration of DETA(0.3-1.2 m mol/L),compared with negative group cell proliferation is reduced,the MDA level significantly increased(P<0.05),cell apoptosis rate increased significantly(P<0.01).C onclusion It is recommended that 0.9 mmol/L of DETA pretreat glial cells for 12 h.The damage to the cells is mild,and the HIF-1αprotein level and mRNA expression are relatively high.It is the ideal concentration of nitric oxide-induced Hif-1αmodel.
作者
牛莉莉
阎萍
李建明
NIU Lili;YAN Ping;LI Jianming(Department of clinical Laboratory,People’s Hospital of Changji Hui Autonomous Prefecture,Changji,Xinjiang,831100,China)
出处
《新疆医学》
2022年第6期613-618,共6页
Xinjiang Medical Journal
基金
新疆维吾尔自治区自然科学基金(项目编号:2020D01A03)
