糖尿病和糖尿病肾病患者循环lncRNA表达谱的分析

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目的分析健康对照组、糖尿病患者、糖尿病肾病患者循环lnc RNA及m RNA表达谱的差异,探讨糖尿病和糖尿病肾病的表观遗传学发病机制,为糖尿病肾病寻找新的治疗靶点提供依据。方法采集2015年1月至3月中国医科大学附属第一医院肾内科收治的肾活检病理明确为结节性糖尿病肾小球硬化症患者21例(DN组)、2型糖尿病患者9例(DM组)及19名正常健康人(N组)的血清。采用Arraystar Human lnc RNA/m RNA V3.0芯片进行lnc RNA和m RNA表达谱检测,使用Agilent Feature Extraction软件对获得的微阵列图像包含的讯息进行提取,采用Gene Spring GX软件包对获得lnc RNA及m RNA原始表达讯息进行标准化。结果与糖尿病组及正常对照组相比,糖尿病肾病组患者尿白蛋白/肌酐比值和血清肌酐显著升高,估计肾小球滤过率(estimated glomeruar filtration rate,e GFR)显著下降(P<0.05)。与健康对照组相比,糖尿病组血清中有245个lnc RNA表达上调和680个lnc RNA表达下调。与糖尿病组相比,糖尿病肾病组血清中有45个lnc RNA表达上调和813个lnc RNA表达下调。自正常组到糖尿病组,再到糖尿病肾病组,lnc RNA-ARAP1-AS2表达逐渐上调(DM/N 2.82倍,DN/DM 2.47倍),lnc RNAARAP1-AS1表达逐渐下调(DM/N 2.24倍,DN/DM 4.79倍),其靶基因ARAP1(Arf GAP with Rho GAP domain,ankyrin repeat and PH domaim 1)m RNA的表达逐渐上调(DM/N 2.25倍,DN/DM 2.45倍)。结论表达下调的lnc RNA-ARAP1-AS1和表达上调的lnc RNA-ARAP1-AS2作用于靶基因m RNA-ARAP1,使m RNA-ARAP1在糖尿病和糖尿病肾病中表达上调,可能通过调控Rho激酶和EGF受体途径,参与了糖尿病和糖尿病肾病的发生,其有望成为糖尿病和糖尿病肾病新的生物标志物。 Objective To analyze the difference of circulating lnc RNA and m RNA expression between healthy control group, diabetic patients and diabetic nephropathy patients, explore the epigenetic mechanism of diabetes mellitus and diabetic nephropathy and provide basis for finding new therapeutic targets for diabetic nephropathy. Methods The pathology of renal biopsy admitted to Department of Nephrology, the First Affiliated Hospital of China Medical University from January 2015 to March 2015 was identified as nodular diabetic glomerulosclerosis in 21 cases (DN group), type 2 diabetes mellitus in 9 cases (DM Group) and 19 normal healthy people (group N). Lnc RNA and m RNA expression profiling were performed using Arraystar Human lnc RNA / m RNA V3.0 chip. The information contained in the obtained microarray images was extracted using Agilent Feature Extraction software. Gene Spring GX software package was used to obtain lnc RNA and The original m RNA expression message was normalized. Results Compared with diabetic group and normal control group, urinary albumin / creatinine ratio and serum creatinine were significantly increased in diabetic nephropathy group, and the estimated glomerular filtration rate (eGFR) was significantly decreased (P <0.05 ). Compared with the healthy control group, there were 245 lnc RNAs and 680 lnc RNAs in the serum of diabetic patients. Compared with the diabetic group, 45 lnc RNA and 813 lnc RNA were down-regulated in the serum of diabetic nephropathy group. The expression of lnc RNA-ARAP1-AS2 was gradually up-regulated (2.82-fold in DM / N and 2.47-fold in DN / DM) from the normal group to the diabetic group and then to the diabetic nephropathy group. DN / DM 4.79-fold), the target gene ARAP1 (Arf GAP with Rho GAP domain, ankyrin repeat and PH domaim 1) m RNA expression was gradually increased (DM / N 2.25 times, DN / DM 2.45 times). Conclusions The downregulation of lnc RNA-ARAP1-AS1 and upregulated lnc RNA-ARAP1-AS2 on the target gene m RNA-ARAP1 up-regulated the expression of m RNA-ARAP1 in diabetic and diabetic nephropathy, possibly through the regulation of Rho kinase and EGF Receptor pathway involved in the development of diabetes and diabetic nephropathy, which is expected to become a new biomarker of diabetes and diabetic nephropathy.
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