急性肺损伤模型中Sdr9c7基因的作用研究

Changes of Sdr9c7 Gene Expression in Acute Lung Injury Model

检测了经鼻腔滴注脂多糖(LPS)溶液诱导的急性肺损伤模型小鼠肺组织Sdr9c7基因表达变化,并探讨其在急性肺损伤的作用。取9只雌性C57BL/6小鼠随机分为空白组、第3天组和第7天组,每组3只。第3天组和第7天组小鼠用LPS诱导急性肺损伤模型,第3天组于LPS诱导的第3天终止实验,第7天组于LPS诱导的第7天终止实验,经鼻腔滴注PBS溶液作对照空白组。转染SDR9C7-siRNA至A549细胞并检测敲低表达的效果。采用苏木精-伊红(HE)染色观察小鼠肺组织形态学,并评估病理评分;用实时荧光定量PCR(RT-qPCR)检测各组肺组织和细胞的IL-1β、TNF-α、IL-6和Sdr9c7基因相对表达量。与空白组比较,第3天组和第7天组肺水肿评分、炎症评分、总病理评分以及Sdr9c7、IL-1β、TNF-α和IL-6基因相对表达量均升高(均P<0.05);与第3天组比较,第7天组炎症评分、总病理评分以及Sdr9c7、IL-1β、TNF-α基因相对表达量降低(均P<0.05)。小鼠肺组织Sdr9c7基因相对表达量与病理评分呈正相关(r=0.964,P<0.01)。敲低Sdr9c7基因实验中,SDR9C7-siRNA3效果更明显。SDR9C7-siRNA3敲低Sdr9c7基因对LPS诱导A549细胞影响实验中,RT-qPCR检测IL-1β、TNF-α、IL-6、Sdr9c7基因相对表达量,与对照组比较,诱导组升高(均P<0.05),敲低组和联合组下降(均P<0.05),且敲低组低于联合组(均P<0.05)。结果表明,急性肺损伤模型小鼠肺组织Sdr9c7基因的异常表达与肺损伤病理评分呈正相关,肺损伤机制可能与Sdr9c7基因高表达促进炎症反应作用相关。

To observe the expression of Sdr9c7 gene in the lung tissue of mice with acute lung injury(ALI)and explore its mechanism.9 female C57BL/6 mice were randomly divided into blank group,3rd-day group and 7th-day group,with 3 mice in each group.Acute lung injury model was induced by lipopolysaccharide(LPS).The experiment was terminated on the 3rd day or on the 7th day of LPS induction respectively.PBS solution was intranasal instilled in the blank group as control group.A549 cells were transfected with SDR9C7-siRNA and the effect of knockdown of Sdr9c7 gene expression was verified.A549 cells were divided into control group,induction group,knockdown group and combination group.Hematoxylin eosin(HE)staining was used to observe the morphology of lung tissue and evaluate the pathological score.The relative expression levels of IL-1β,TNF-α,IL-6 and Sdr9c7 genes in lung tissue and cells were detected by real-time quantitative PCR(RT-qPCR).Results showed that compared with the blank group,the pulmonary edema score,inflammation score,total pathological score and the relative expression of Sdr9c7,IL-1β,TNF-αand IL-6 genes in the 3rd and 7th day groups were increased(all P<0.05);compared with the 3rd day group,the inflammation score,total pathological score and the relative expression of Sdr9c7,IL-1βand TNF-αgenes in the 7th day group were decreased(all P<0.05).The relative expression of Sdr9c7 gene was positively correlated with pathological score(r=0.964,P<0.01).In the experiment of knockdown of Sdr9c7 gene,the effect of SDR9C7-siRNA3 was more significant.In the experiment of SDR9C7-siRNA3 knockdown in LPS induced A549 cells,the relative expression levels of IL-1β,TNF-α,IL-6 and Sdr9c7 were detected by RT-qPCR.Compared with the control group,the expression levels of IL-1β,TNF-α,IL-6 and Sdr9c7 increased in the induction group(all P<0.05),decreased in the knockdown group and combination group(all P<0.05),and the expression levels of IL-1β,TNF-α,IL-6 and Sdr9c7 in the knockdown group were lower than those in the combination group(all P<0.05).It suggests that the abnormal expression of Sdr9c7 gene in lung tissue of acute lung injury model mice may be related to LPS induced inflammatory response.