Evaluation of perilipin expression in the kidney tissues of rats with diabetic nephropathy
Received:November 26, 2018  
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DOI:10. 3969 / j.issn.1005-4847. 2019. 03. 012
KeyWord:perilipin; urinary laminin; diabetic nephropathy; 24-hour urine protein; kidneys; pathology; rat
              
AuthorInstitution
卫兵艳 山西医科大学实验动物中心,实验动物与人类疾病动物模型山西省重点实验室,太原
樊林花 山西医科大学实验动物中心,实验动物与人类疾病动物模型山西省重点实验室,太原
刘茂林 山西医科大学实验动物中心,实验动物与人类疾病动物模型山西省重点实验室,太原
轩瑞晶 山西医科大学实验动物中心,实验动物与人类疾病动物模型山西省重点实验室,太原
刘田福 山西医科大学实验动物中心,实验动物与人类疾病动物模型山西省重点实验室,太原
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Abstract:
      Objective To characterize the rat model of diabetic nephropathy (DN), to evaluate the diagnosticvalue of urinary laminin for early detection of DN, and to investigate the expression of perilipin in diabetic rat kidneys.Methods Fourteen male Sprague-Dawley rats were randomly divided into a control group (regular diet, six animals) anda diabetic nephropathy model group (high sugar and high fat diet, eight animals). After feeding for 4 weeks, the rats of thedisease model group were injected with a dose of 30 mg/ kg of 1% streptozotocin. Induction of diabetes was consideredsuccessful when blood sugar levels were ≥ 16. 7 mmol/ L. Upon induction of diabetes, animals were fed for an additional 6weeks. Induction of diabetic nephropathy was considered successful when the 24-hour urinary protein level was ≥ 30 mg/ kg.Coomassie brilliant blue (CBB) was used to determine the 24-hour urine protein levels, ELISA was used to measure urinelaminin, and hematoxylin and eosin (H&E) staining was performed to observe the pathological changes in kidney tissues.Perilipin (Plin) expression in kidney tissues was determined by real-time PCR and western blotting. Results Thedetection of 24-hour urinary protein levels ≥ 30 mg/ kg confirmed the successful induction of the rat model of diabeticnephropathy. The kidney-to-body weight ratio of the disease model group was increased significantly, when compared withthe control diet group ( P <0. 05). Urinary volume and laminin increased by the 5th week, while 24-hour urine proteinincreased by the 6th week. All the three indicators were increasing over time. Pathological examination of the renal tissuesrevealed glomerular hypertrophy, basal membrane hyperplasia, microhemangioma formation, tubular cavity deformation,epithelial shedding and vacuolization, inflammatory monocyte and lymphocyte infiltration, and interstitial collagendeposition. A significant increase in Plin expression at the mRNA and protein levels in the kidney tissues of diabeticnephropathy rats was also observed. Conclusions Urinary laminin is increased earlier than the 24-hour urine protein level,thus can be used as an early biomarker of diabetic nephropathy. Increased Plin expression may play a role in thepathogenesis of diabetic nephropathy, and this protein therefore warrants further investigation to acquire a better understanding the molecular mechanisms underlying this disease.
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