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卫兵艳,樊林花,刘茂林,轩瑞晶,刘田福.周脂素在糖尿病肾病大鼠肾组织中的表达[J].中国实验动物学报,2019,27(3):347~352.
周脂素在糖尿病肾病大鼠肾组织中的表达
Evaluation of perilipin expression in the kidney tissues of rats with diabetic nephropathy
投稿时间:2018-11-26  
DOI:10. 3969 / j.issn.1005-4847. 2019. 03. 012
中文关键词:  周脂素  尿层粘连蛋白  糖尿病肾病  24 h 尿蛋白  肾脏  病理学  大鼠
英文关键词:perilipin  urinary laminin  diabetic nephropathy  24-hour urine protein  kidneys  pathology  rat
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作者单位E-mail
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 030001 280467659@ qq.com 
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 030001  
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 030001  
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 030001  
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 030001 13603518575@ 163.com 
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中文摘要:
      目的 观察大鼠糖尿病肾病(diabetic nephropathy,DN)模型的特点和尿LN 对早期DN 的诊断价值,探讨周脂素(perilipin,Plin)在DN 大鼠肾脏中的表达情况?方法 将14 只SD 雄性大鼠随机分为对照组(普通饲料)和糖尿病肾病模型组(高糖高脂饲料),对照组6 只,模型组8 只,饲养4 周后模型组按照30 mg/ kg 剂量注射1%链脲佐菌素(STZ),检测血糖≥16. 7 mmol/ L,糖尿病模型制作成功,继续喂养6 周,检测24 h 尿蛋白≥30 mg/ kg,糖尿病肾病模型制作成功?考马斯亮蓝检测24 h 尿蛋白?ELISA 测尿层粘连蛋白,HE 染色观察肾组织的病理变化,Real-time PCR 及Western blot 检测肾脏组织中perilipin 表达情况?结果 模型鼠24 h 尿蛋白≥30 mg/ kg,糖尿病肾病大鼠模型制作成功?和对照组大鼠相比,模型组的肾重/ 体重比明显增高( P < 0. 05),尿量?尿层粘连蛋白于5 周出现升高?24 h 尿蛋白于6 周时出现升高,且三项指标均随着时间不断增高?肾组织病理检查显示:肾小球肥大,基膜增生, 微小血管瘤形成,肾小管管腔变形,上皮脱落?空泡样变,大量单核?淋巴等炎性细胞浸润,间质内胶原纤维增生?模型组大鼠肾组织Plin 的mRNA 及蛋白表达均明显的升高( P < 0. 05)?结论 尿层粘连蛋白比24 h尿蛋白升高得早,可作为早期糖尿病肾病的警示指标?Plin 表达增高可能参与了糖尿病肾病肾病变过程,为进一步探讨糖尿病肾病的发病机制提供新的思路?
英文摘要:
      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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