摘要
目的探讨微小RNA-34a（miR-34a）是否通过调控Kruppel样因子4（KLF4）参与脂多糖（LPS）介导的脓毒症相关性肾损伤。方法30只健康雄性SD大鼠，体重180~200 g，按随机数字表法分为对照组和模型组，每组15只。模型组大鼠经尾静脉注射LPS 7.5 mg/kg诱导脓毒症相关性肾损伤模型；对照组大鼠则注射等量生理盐水。用多功能生化仪检测两组大鼠血肌酐（SCr）和尿素氮（BUN）含量；经苏木素-伊红（HE）染色后观察肾组织病理学改变；用实时荧光定量反转录-聚合酶链反应（qRT-PCR）检测血浆和肾组织miR-34a及KLF4的基因表达；用蛋白质免疫印迹试验（Western Blot）检测肾组织KLF4蛋白表达；用双荧光素酶报告基因分析法验证KLF4是否为miR-34a靶基因。结果与对照组比较，模型组肾组织炎性细胞浸润增加，血SCr和BUN显著升高〔SCr（μmol/L）：142.5±10.6比46.4±5.6，BUN（mmol/L）：31.6±6.2比8.5±1.2，均P〈0.01〕，血浆和肾组织miR-34a基因表达明显增加（2 -Ct：血浆为2.26±0.11比1.14±0.05，肾组织为4.23±0.12比1.12±0.04，均P〈0.01），肾组织KLF4基因和蛋白表达明显降低基因表达（2 -Ct）：0.52±0.03比1.21±0.06，蛋白表达（A值）：0.72±0.03比1.05±0.04，均P〈0.01〕，说明大鼠发生了肾损伤。Pearson相关性分析显示，血浆miR-34a？mRNA表达与SCr和BUN均呈正相关（r值分别为0.678、0.721，均P〈0.01）。双荧光素酶报告基因系统证实，KLF4是miR-34a的靶基因。结论miR-34a通过调控KLF4参与了LPS介导的脓毒症相关性肾损伤。
ObjectiveTo investigate whether microRNA-34a （miR-34a） participates in lipopolysaccharide （LPS） mediated sepsis related renal function impairment via Kruppel-like factor 4 （KLF4）.MethodsThirty healthy male Sprague-Dawley （SD） rats, weighing 180-200 g, were randomly divided into two groups： control group and model group, with 15 rats in each group. The SD rats from model group were injected with LPS 7.5 mg/kg to induce sepsis related renal function impairment model, the SD rats from control group were injected with normal saline. The serum creatinine concentration （SCr） and blood urine nitrogen （BUN） content was detected by multifunction biochemical analyzer; the morphological changes of renal tissue were observed by hematoxylin and eosin stain （HE） staining; the expression of miR-34a and KLF4 gene in plasma and renal tissue were detected by real-time quantitative reverse transcriptase polymerase chain reaction （qRT-PCR）; the protein expression of KLF4 in renal tissue was detected by Western Blot; the target gene of miR-34a was verified by double luciferase reporter gene analysis.ResultsCompared with control group, inflammatory cell infiltration in renal tissue was increased in model group, the SCr and BUN were significantly increased [SCr （μmol/L）： 142.5±10.6 vs. 46.4±5.6, BUN （mmol/L）： 31.6±6.2 vs. 8.5±1.2, bothP〈 0.01], the gene expression of miR-34 in plasma and renal tissue were significantly increased （2 -Ct： 2.26±0.11 vs. 1.14±0.05 in plasma, 4.23±0.12 vs. 1.12±0.04 in renal tissue, bothP〈 0.01）, the gene and protein expressions of KLF4 were significantly decreased [KLF4 gene （2 -Ct？）： 0.52±0.03 vs. 1.21±0.06, KLF4 protein （Avalue）： 0.72±0.03 vs. 1.05±0.04, bothP〈 0.01], which indicated that kidney injury occurred in rats. Pearson correlation analysis showed that plasma miR-34a was positively correlated with SCr and BUN （rvalue were 0.678, 0.721, respectively, bothP〈 0.05）. Double luciferase reporter assay confirmed t
作者
姜启栋
伍长学
张琼
Jiang Qidong;Wu Changxue;Zhang Qiong(Department of Intensive Care Unit, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan, China （Jiang QD, Wu CX;Department of Nephrology, the Second Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan, China （Zhang Q)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2018年第4期351-354,共4页
Chinese Critical Care Medicine
基金
四川省卫生和计划生育委员会课题（17PJ396）
