[微小RNA-34a通过Kruppel样因子4参与脂多糖介导的脓毒症相关肾功能损害]
[microRNA-34a participates in lipopolysaccharide mediated sepsis related renal function impairment via Kruppel-like factor 4].
作者信息
Jiang Qidong, Wu Changxue, Zhang Qiong
机构信息
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). Corresponding author: Zhang Qiong, Email:
出版信息
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2018 Apr;30(4):351-354. doi: 10.3760/cma.j.issn.2095-4352.2018.04.013.
OBJECTIVE
To investigate whether microRNA-34a (miR-34a) participates in lipopolysaccharide (LPS) mediated sepsis related renal function impairment via Kruppel-like factor 4 (KLF4).
METHODS
Thirty 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.
RESULTS
Compared 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, both P < 0.01], the gene expression of miR-34 in plasma and renal tissue were significantly increased (2: 2.26±0.11 vs. 1.14±0.05 in plasma, 4.23±0.12 vs. 1.12±0.04 in renal tissue, both P < 0.01), the gene and protein expressions of KLF4 were significantly decreased [KLF4 gene (2): 0.52±0.03 vs. 1.21±0.06, KLF4 protein (A value): 0.72±0.03 vs. 1.05±0.04, both P < 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 (r value were 0.678, 0.721, respectively, both P < 0.05). Double luciferase reporter assay confirmed that KLF4 was the target gene of miR-34a.
CONCLUSIONS
The miR-34a participates in LPS mediated sepsis related renal function impairment via KLF4.
目的
探讨微小RNA-34a（miR-34a）是否通过 Kruppel样因子4（KLF4）参与脂多糖（LPS）介导的脓毒症相关肾功能损害。
方法
将30只体重180 - 200 g的健康雄性Sprague-Dawley（SD）大鼠随机分为两组：对照组和模型组，每组15只。模型组SD大鼠注射7.5 mg/kg LPS诱导脓毒症相关肾功能损害模型，对照组SD大鼠注射生理盐水。用多功能生化分析仪检测血清肌酐浓度（SCr）和血尿素氮（BUN）含量；通过苏木精-伊红染色（HE染色）观察肾组织的形态学变化；用实时定量逆转录聚合酶链反应（qRT-PCR）检测血浆和肾组织中miR-34a和KLF4基因的表达；用蛋白质免疫印迹法检测肾组织中KLF4的蛋白表达；通过双荧光素酶报告基因分析验证miR-34a的靶基因。
结果
与对照组相比，模型组肾组织中炎性细胞浸润增加，SCr和BUN显著升高[SCr（μmol/L）：142.5±10.6 vs. 46.4±5.6，BUN（mmol/L）：31.6±6.2 vs. 8.5±1.2，均P < 0.01]，血浆和肾组织中miR-34的基因表达显著增加（血浆中：2.26±0.11 vs. 1.14±0.05，肾组织中：4.23±0.12 vs. 1.12±0.04，均P < 0.01），KLF4的基因和蛋白表达显著降低[KLF4基因（2）：0.52±0.03 vs. 1.21±0.06，KLF4蛋白（A值）：0.72±0.03 vs. 1.05±0.04，均P < 0.01]，表明大鼠发生了肾损伤。Pearson相关性分析显示血浆miR-34a与SCr和BUN呈正相关（r值分别为0.678、0.721，均P < 0.05）。双荧光素酶报告基因检测证实KLF4是miR-34a的靶基因。
结论
miR-34a通过KLF4参与LPS介导的脓毒症相关肾功能损害。
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