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目的:评估帕瑞昔布对过氧化氢诱导的大鼠原代星形胶质细胞氧化应激状态的保护作用,并初步探讨其作用机制。创新点:首次在大鼠原代星形胶质细胞中证实帕瑞昔布对过氧化氢诱导的氧化应激具有保护作用,且此作用可能与Bax、Bcl-2和BDNF的失调有关。方法:设立如下四组对照:(1)阴性对照组;(2)100μmol/L H_2O_2处理组(处理时间为24 h);(3)80μmol/L帕瑞昔布处理组;(4)160μmol/L帕瑞昔布处理组(第三和第四组先用帕瑞昔布处理24 h,再用100μmol/L H_2O_2处理24 h)。用荧光显微镜观察星形胶质细胞的形态,用MTT法检测星形胶质细胞的存活率,用荧光探针二氯荧光黄双乙酸盐(DCDHF-DA)检测星形胶质细胞内氧自由基的含量,并用碘化丙啶(PI)染色检测细胞的凋亡状态。最后用反转录酶聚合酶链反应(RT-PCR)和蛋白质印迹(Western blot)检测Bax、Bcl-2和BDNF三种蛋白在4组中的表达水平。结论:H_2O_2处理可以导致星形胶质细胞的形态发生改变(图1)和存活率降低(图2),提高星形胶质细胞内的氧自由基水平(图3),同时诱导细胞凋亡(图4)。然而,所有这些变化都可以被帕瑞昔布逆转。此外,我们发现,Bax、Bcl-2和BDNF的表达水平在H_2O_2处理时失调,而在帕瑞昔布预处理时恢复正常。综上所述,帕瑞昔布对H_2O_2诱导的氧化应激具有保护作用。
PURPOSE: To evaluate the protective effect of parecoxib against hydrogen peroxide-induced oxidative stress in rat primary astrocytes and to explore its mechanism. Innovation: For the first time in primary rat astrocytes confirmed that parecoxib has a protective effect against hydrogen peroxide-induced oxidative stress, and this effect may be related to the imbalance of Bax, Bcl-2 and BDNF. METHODS: The following four groups of controls were set up: (1) negative control group; (2) 100 μmol / L H 2 O 2 treatment group for 24 h; (3) 80 μmol / L parecoxib treatment group; L parecoxib treatment group (groups 3 and 4 were treated with parecoxib for 24 h followed by 100 μmol / L H 2 O 2 for 24 h). The morphology of astrocytes was observed by fluorescence microscopy. The viability of astrocytes was detected by MTT assay. The content of oxygen free radicals in astrocytes was detected by fluorescent probe dichloro-fluorescein diacetate (DCDHF-DA) , And the apoptosis status of the cells was detected by propidium iodide (PI) staining. Finally, the expression of Bax, Bcl-2 and BDNF in four groups were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot. CONCLUSION: H 2 O 2 treatment results in altered morphology of astrocytes (Figure 1) and decreased survival (Figure 2), increased levels of oxygen free radicals in astrocytes (Figure 3), and induced apoptosis (Figure 4). However, all these changes can be reversed by parecoxib. In addition, we found that the expression levels of Bax, Bcl-2 and BDNF were deregulated during H 2 O 2 treatment and returned to normal after pretreatment with parecoxib. In summary, parecoxib has a protective effect on H 2 O 2 induced oxidative stress.