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目的观察烧伤小鼠干扰素诱导表达蛋白IFIT1的表达变化,初步探究其表达变化的原因。方法实验小鼠致TBSA15%Ⅱ度烧伤,1d及7d后处死,取肝、肺及脾组织。脂多糖掺入小鼠腹腔巨噬细胞株RAW264.7的培养基中,至终浓度0.1~1.0μg/ml,作用6h。提取组织及细胞RNA,进行半定量逆转录-聚合酶链式反应,提取组织及细胞裂解上清,进行免疫印迹检测。结果小鼠肝、肺、脾组织IFIT1的转录在烧伤后1d升高,伤后7d恢复至伤前水平;肝、肺、脾IFIT1的蛋白水平在伤后1d一致升高,伤后7d仍能检出升高。体外实验,细菌脂多糖显著激活RAW264.7细胞的IF-IT1转录及蛋白表达。结论烧伤小鼠伤后早期IFIT1表达迅速升高,此变化与同期的内毒素血症引起的细胞应激有关。
Objective To investigate the expression of interferon-inducible protein (IFIT1) in burn mice and to explore the possible causes of the changes. Methods TBSA was induced in 15% of second degree burn mice and sacrificed on day 1 and day 7 after transplantation. The liver, lung and spleen tissues were harvested. Lipopolysaccharide was incorporated into the medium of mouse peritoneal macrophage cell line RAW264.7 to a final concentration of 0.1-1.0 μg / ml for 6 hours. Tissues and cell RNAs were extracted and subjected to semi-quantitative reverse transcriptase-polymerase chain reaction (PCR). Tissues and cell lysates were extracted and subjected to Western blotting. Results The transcription of IFIT1 in the liver, lung and spleen of mice increased at 1 day after burn, and recovered to the pre-injury level 7 days after injury. The protein level of IFIT1 in liver, lung and spleen increased consistently at 1 day after injury, Checked up. In vitro, bacterial lipopolysaccharide significantly activated IF-IT1 transcription and protein expression in RAW264.7 cells. Conclusion The expression of IFIT1 in burned mice rapidly increases after burn injury, which is related to the cellular stress induced by endotoxemia in the same period.