肾缺血再灌注致肺损伤关键基因Alox5ap的筛选及其表达与功能验证  

Screening of Key Gene Alox5ap for Renal Ischemia-Reperfusion-Caused Lung Injury and Its Expression and Function Verification

作　　者：欧阳秀芳 赵宁[1] 付尚淼 孙智坚 钱克俭[1] 陶文强 OUYANG Xiu-fang;ZHAO Ning;FU Shang-miao;SUN Zhi-jian;QIAN Ke-jian;TAO Wen-qiang(Department of Critical Care Medicine,the 1 st Affiliated Hospital,Jiangxi Medical College,Nanchang University,Nanchang,330006,China)

机构地区：[1]南昌大学第一附属医院重症医学科,南昌330006

出　　处：《南昌大学学报（医学版）》2023年第6期8-14,共7页Journal of Nanchang University：Medical Sciences

基　　金：江西省自然科学基金(20232BAB206083);江西省卫健委科技计划项目(202210371);南昌大学第一附属医院青年人才科研培育基金(YFYPY202264)。

摘　　要：目的 筛选肾缺血再灌注(RIR)致肺损伤的关键基因并对其表达与功能进行验证。方法 GEO数据库下载RIR致肺损伤数据集GSE6730,利用生物信息学方法筛选关键基因。SD大鼠按照随机数字表法分为2组:假手术组(Sham组)和RIR组,每组6只;HE染色观察2组大鼠肺组织病理形态;qRT-PCR检测2组大鼠肺组织中TNF-α、IL-6、IL-18及花生四烯酸5-脂氧合酶激活蛋白(Alox5ap) mRNA的表达。常规培养大鼠肺泡Ⅱ型上皮细胞RLE-6TN,将其分为Control组、LPS组(LPS处理)、LPS+si-NC组(转染si-NC联合LPS处理)、LPS+si-Alox5ap组(转染si-Alox5ap联合LPS处理)、LPS+MK-886组(LPS联合抑制剂MK-886处理);qRT-PCR检测各组TNF-α、IL-6、IL-18的mRNA表达。结果 Alox5ap为RIR致肺损伤的关键基因。与Sham组比较,RIR组肺组织见明显损伤,TNF-α、IL-6、IL-18及Alox5ap mRNA表达均增加(P<0.05)。与Control组比较,LPS组TNF-α、IL-6、IL-18 mRNA表达均升高(P<0.05);与LPS+si-NC组比较,LPS+si-Alox5ap组的TNF-α、IL-6、IL-18的mRNA表达均降低(P<0.05);与LPS组比较,LPS+MK-886组的TNF-α、IL-6、IL-18的mRNA表达均降低(P<0.05)。结论 成功筛选RIR致肺损伤关键基因Alox5ap,RIR致肺损伤后肺组织中Alox5ap表达升高;下调Alox5ap表达可减轻LPS诱导肺损伤的炎症因子表达。Objective To screen the key genes for lung injury induced by renal ischemia-reperfusion(RIR),and to verify their expression and function.Methods The dataset GSE6730 of RIR-induced lung injury was downloaded from Gene Expression Omnibus(GEO) database,and the key genes were screened by bioinformatics analysis.Twelve SD rats were randomly divided into sham group and RIR group,with 6 rats in each group.Pathological changes in the lung tissue were observed by H&E staining.The mRNA expression ofarachidonic acid 5-lipoxygenase-activated protein(Alox5ap) in the lung tissue was determined by qRT-PCR.Rat alveolar type Ⅱ epithelial cells(RLE-6TN) were routinely cultured,and divided into control group,lipopolysaccharide(LPS) group(cells were treated with LPS),LPS+si-NC group(cells were transfected with si-NC and treated with LPS),LPS+si-Alox5ap group(cells were transfected with si-Alox5ap and treated with LPS),and LPS+MK-886 group(cells were treated with LPS and MK-886).The mRNA expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-18 in the lung tissue and cells were detected by qRT-PCR.Results Alox5ap was screened as a key gene in lung injury induced by RIR.Compared with the sham group,lung tissue was obviously damaged and the expression levels of TNF-α,IL-6,IL-18 and Alox5ap were significantly increased in the RIR group(P<0.05).The expression levels of TNF-α,IL-6 and IL-18 in the LPS group was higher than those in the control group,but those in the LPS+si-Alox5ap group were lower than those in the LPS+si-NC group(P<0.05).In addition,the expression levels of TNF-α,IL-6 and IL-18 in the LPS+MK-886 group were lower than those in the LPS group(P<0.05).Conclusion The key gene Alox5ap for RIR-caused lung injury was successfully screened,and Alox5ap expression increased in the lung tissue after lung injury induced by RIR.The downregulation of Alox5ap expression could reduce the expression of inflammatory factors in LPS-induced lung injury.

关 键 词：肺损伤 肾缺血再灌注 生物信息学 花生四烯酸5-脂氧合酶激活蛋白 动物 实验 大鼠 

分 类 号：R629.2[医药卫生—整形外科]