尿外泌体中致纤维化基因表达在早期糖尿病肾病诊断中的作用研究

The Role of Fibrogenic Gene Expression in Urinary Exosomes in the Diagnosis of Early Diabetic Nephropathy

目的 探讨尿外泌体中致纤维化基因表达在诊断早期糖尿病肾病的应用价值。方法 选取2019年5月—2020年6月于广东医科大学附属厚街人民医院住院的新诊断早期2型糖尿病肾病患者16例及2型糖尿病肾病患者16例,将32例患者分别纳入研究组A组(早期2型糖尿病肾病组)和B组(2型糖尿病肾病组)。同时选取同期到医院进行体检的健康体检者16名,纳入对照组。对高糖刺激肾小管上皮外泌体miRNA进行系统分析(芯片检测)。收集了两组DN患者肾穿刺标本,进行组化染色,对比两组患者血肌酐、尿白蛋白、尿钠等的水平以及对比两组患者尿外泌体中转化生长因子-β1(TGF-β1)和结缔组织生长因子(CTGF)的mRNA的表达等相关情况。结果 研究组血肌酐、尿白蛋白、尿钠的水平明显高于对照组,差异有统计学意义(P<0.05);研究组A组的血肌酐、尿白蛋白、尿钠的水平低于研究组B组,差异有统计学意义(P<0.05)。研究组A组和B组患者尿液中的TGF-β1和CTGF的mRNA表达水平均明显高于对照组(2.2±0.8)、(4.1±1.2)vs(1.0±0.2)、(1.8±1.1)、(3.8±1.8)vs(1.1±0.3),差异有统计学意义(P<0.05)。研究B组患者尿液中的TGF-β1和CTGF的mRNA表达水平明显高于研究A组,分别为(4.1±1.2)vs(2.2±0.8)和(3.8±1.8)vs(1.8±1.1),差异有统计学意义(P<0.05)。结论 糖尿病肾病患者尿外泌体中致纤维化基因TGF-β1和CTGF可能是临床诊断早期糖尿病肾病及其进展的重要辅助生物学标志物。

Objective To explore the application value of fibrotic gene expression in urinary exosomes in the diagnosis of early diabetic nephropathy.Methods A total of 16 newly diagnosed patients with early type 2 diabetic nephropathy and 16 patients with type 2 diabetic nephropathy who were hospitalized in Houjie People’s Hospital Affiliated to Guangdong Medical University from May 2019 to June 2020 were selected.32 patients were included in the study group A(early type 2 diabetic nephropathy group) and group B(type 2 diabetic nephropathy group).At the same time,16 healthy subjects who went to the hospital for physical examination during the same period were selected and included in the control group.Systematic analysis(chip assay) of high glucose-stimulated renal tubular epithelial exosomal miRNAs.The renal puncture specimens of the two groups of DN patients were collected,and histochemical staining was performed to compare the levels of serum creatinine,urinary albumin,and urinary sodium in the two groups of patients.The mRNA expressions of transforming growth factor-β1(TGF-β1) and connective tissue growth factor(CTGF) in urinary exosomes of the two groups were compared.Results According to the survey results,the levels of serum creatinine,urinary albumin and urinary sodium in the study group were significantly higher than those in the control group,and the difference was statistically significant(P<0.05).The levels of serum creatinine,urine albumin,and urine sodium in group A were lower than those in group B,and the difference was statistically significant(P<0.05).The results showed that the mRNA expression levels of TGF-β1and CTGF in the urine of the patients in the study group A and B were significantly higher than those in the control group(2.2±0.8),(4.1±1.2) vs(1.0±0.2);(1.8±1.1),(3.8±1.8) vs(1.1±0.3),and the difference was statistically significant(P<0.05).Moreover,the mRNA expression levels of TGF-β1and CTGF in the urine of patients in study group B were significantly higher than those in study group A,(4.1±1.2) vs(2.2±0.8) and(3.8±1.8) vs(1.8±1.1) and the difference was statistically significant(P<0.05).Conclusion The fibrotic genes TGF-β1and CTGF in the urinary exosomes of patients with diabetic nephropathy may be important auxiliary biomarkers for clinical diagnosis of early diabetic nephropathy and its progression.