Li Lei, Song Hai-Yan, Liu Kai, An Meng-Meng
Department of Endocrinology and Metabolism, The Second Affiliated Hospital of Harbin Medical University, Hormone and Endocrinology Key Laboratory of Harbin Medical University Xuefu Road 246, Harbin 150080, China.
Int J Clin Exp Med. 2015 Aug 15;8(8):12746-55. eCollection 2015.
To detect the serum concentrations of secreted protein acidic and rich in cysteine (SPARC) in patients with diabetic nephropathy and SPARC mRNA and protein expressions in renal tissue of db/db mice (C57BL/KsJ, diabetic nephropathy mice), thus preliminary exploration on the role of secreted protein acidic riches in cysteine in the development of diabetic nephropathy were carried out. Serum SPARC levels in normal subjects, patients with type 2 diabetes mellitus (without diabetic nephropathy), chronic renal failure (without diabetes mellitus), and diabetic nephropathy were determined with enzyme-linked immunosorbent assay. 12-week-old db/db mice (db/db group) and its littermate wild-type control mice (NC group) were selected with 6 from each group, and the kidney tissue were taken. RT-PCR, Western blot, and immunofluorescence were used to detect the mRNA, targeted protein expressions of SPARC and the staining of renal tissue. The serum level of SPARC in diabetic nephropathy group was significantly higher than those in normal group, type 2 diabetes mellitus, and chronic renal failure group (P < 0.05 or P < 0.01). The SPARC level in the type 2 diabetes mellitus group was higher than that in normal group (P < 0.05), but there was no difference between normal group and chronic renal failure. SPARC mRNA and protein levels in renal tissue of db/db mice were higher compared with the normal control group (P < 0.05). The long term hyperglycemic state in patients with diabetic nephropathy causes pathological change of renal tissue. Simultaneously, increased secretion of SPARC from renal tissue results in elevation of serum SPARC level. SPARC correlates with the occurrence and progression of diabetes, and it may play a role in pathological change of diabetic nephropathy.
检测糖尿病肾病患者血清中富含半胱氨酸的酸性分泌蛋白(SPARC)浓度以及db/db小鼠(C57BL/KsJ,糖尿病肾病小鼠)肾组织中SPARC mRNA和蛋白表达,从而初步探讨富含半胱氨酸的酸性分泌蛋白在糖尿病肾病发生发展中的作用。采用酶联免疫吸附测定法检测正常受试者、2型糖尿病患者(无糖尿病肾病)、慢性肾衰竭患者(无糖尿病)及糖尿病肾病患者血清中的SPARC水平。选取12周龄的db/db小鼠(db/db组)及其同窝野生型对照小鼠(NC组),每组6只,取其肾组织。采用RT-PCR、蛋白质免疫印迹法及免疫荧光法检测SPARC的mRNA、靶向蛋白表达及肾组织染色情况。糖尿病肾病组血清SPARC水平显著高于正常组、2型糖尿病组及慢性肾衰竭组(P<0.05或P<0.01)。2型糖尿病组SPARC水平高于正常组(P<0.05),但正常组与慢性肾衰竭组之间无差异。db/db小鼠肾组织中SPARC mRNA和蛋白水平高于正常对照组(P<0.05)。糖尿病肾病患者长期高血糖状态导致肾组织发生病理改变。同时,肾组织中SPARC分泌增加导致血清SPARC水平升高。SPARC与糖尿病的发生发展相关,可能在糖尿病肾病的病理改变中发挥作用。