论文部分内容阅读
目的观测脂多糖(LPS)是否诱导人牙周膜成纤维细胞(HPDLF)发生内质网应激。方法体外培养HPDLF,分别用LPS(0.1、1、10μg/mL)刺激0、3、6、9 h后收集细胞,提取RNA。采用实时荧光定量PCR检测葡萄糖调节蛋白78(GRP78)、CHOPmRNA表达水平,通过RT-PCR检测XBP1mRNA表达水平及剪切活化情况。结果 LPS作用HPDLF 3 h后GRP78、CHOP、XBP1mRNA表达水平显著上调,其中XBP1mRNA表达水平在6 h达到高峰,并且出现活化的剪切形式XBP1s,而GRP78、CHOPmRNA表达水平持续上调。结论 LPS作用HPDLF后细胞发生内质网应激,且存在浓度和一定的时间依赖性。
Objective To observe whether lipopolysaccharide (LPS) induces endoplasmic reticulum stress in human periodontal ligament fibroblasts (HPDLF). Methods HPDLF was cultured in vitro, and the cells were harvested at 0, 1, 6, and 9 h after stimulation with LPS (0.1, 1, 10 μg / mL) respectively. RNA was extracted. Real-time fluorescence quantitative PCR was used to detect the expression of GRP78 and CHOPmRNA. The expression of XBP1mRNA and the activation of shearing were detected by RT-PCR. Results The expression of GRP78, CHOP and XBP1 mRNA in HPDLF treated with LPS for 3 h was significantly up-regulated. The expression level of XBP1 mRNA peaked at 6 h, and the activated form of XBP1s appeared, while the expression of GRP78 and CHOP mRNA was continuously up-regulated. Conclusion LPS treated with HPDLF endoplasmic reticulum stress occurs, and the concentration and time-dependent manner.