CAJ | 학술논문

目的观察内毒素休克(endotoxic shock,ES)前后大鼠早期肺组织基因表达谱的差异,探讨急性肺损伤可能的分子机制.方法 20只雄性Wistar大鼠随机数字表法分为正常对照组和LPS组,每组10只.尾静脉注射LPS 10 mg/kg制备ES大鼠模型(LPS组),6 h后测定大鼠动脉血氧分压(PaO2).提取大鼠肺组织RNA,利用Affymetrix RAT 230A大鼠全基因组芯片对大鼠肺组织基因表达谱进行检测,并用半定量RT-PCR技术对其中5条基因的表达水平进行验证.根据差异基因的类型结合ES的特点分析数据.结果与正常对照组比较,LPS组大鼠PaO2显著下降(P＜0.05);按照显著差异基因的标准,在大鼠15 650条靶基因中,初步筛选出158条差异表达基因,其中上调基因117条,下调基因34条.根据基因的生物学功能,差异表达基因主要为:炎症相关基因、物质转运相关基因、转录调节相关基因、信号转导相关基因、应激反应相关基因、代谢相关基因、细胞凋亡相关基因、细胞黏附相关基因等.其中5条基因的RT-PCR验证结果与芯片分析结果相符.结论 ES大鼠损伤的肺组织中多种基因表达发生变化,其中炎症相关基因的差异表达最为突出.
목적 관찰내독소휴극(endotoxic shock,ES)전후대서조기폐조직기인표체보적차이,탐토급성폐손상가능적분자궤제.방법 20지웅성Wistar대서수궤수자표법분위정상대조조화LPS조,매조10지.미정맥주사LPS 10 mg/kg제비ES대서모형(LPS조),6 h후측정대서동맥혈양분압(PaO2).제취대서폐조직RNA,이용Affymetrix RAT 230A대서전기인조심편대대서폐조직기인표체보진행검측,병용반정량RT-PCR기술대기중5조기인적표체수평진행험증.근거차이기인적류형결합ES적특점분석수거.결과 여정상대조조비교,LPS조대서PaO2현저하강(P＜0.05);안조현저차이기인적표준,재대서15 650조파기인중,초보사선출158조차이표체기인,기중상조기인117조,하조기인34조.근거기인적생물학공능,차이표체기인주요위:염증상관기인、물질전운상관기인、전록조절상관기인、신호전도상관기인、응격반응상관기인、대사상관기인、세포조망상관기인、세포점부상관기인등.기중5조기인적RT-PCR험증결과여심편분석결과상부.결론 ES대서손상적폐조직중다충기인표체발생변화,기중염증상관기인적차이표체최위돌출.
Objective To observe the difference of gene expression profiles of lung in rats before and after endotoxic shock (ES). Methods A total of 20 male Wistar rats were randomly divided into control group and lipopolysaceharide (LPS) group ( 10 rats per group). The LPS rat model was made by injecting LPS into tail vein. Six hours after ES, partial pressure of oxygen in the arterial blood ( PaO2 )was measured. Gene expression profiles of the lung in each group were detected by rats oligo gene chip Affymetrix RAT 230A. The expression level of five genes was verified via semi-quantitative RT-PCR. The data were analyzed in combination with type of differential gene and character of ES. Results Compared with control group, PaO2 in LPS group was decreased more significantly (P ＜0.05). Among 15 650 probes detected, 158 genes showed differential expression in ES group in comparison with control group. The expression level of 117 genes was up-regulated while that of 34 genes down-regulated significantly. According to their biological function, differentially expressed genes were classified as inflammation genes, material transporter genes, transcription regulator genes, signal transduction genes, stress response genes, metabolic genes, apoptosis genes and cell adhesion genes. The results of Semi-quantitative RT-PCR of five genes were consistent with those of the microarray examination. Conclusion The expression of many genes of the lung may change in ES rats, especially the inflammatory genes.