乙醛酸盐诱导小鼠结晶肾损伤模型肾小管上皮细胞早期变化的研究  

Study on early changes of renal tubular epithelial cells induced by glyoxylate in mouse model of crystalline kidney injury

作　　者：张坤 章俊亚 ZHANG Kun;ZHANG Junya(Department of Urology,Zhongda Hospital Lishui Branch,Southeast University,Nanjing 211200,China;Department of Thyroid and Breast Surgery,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,China)

机构地区：[1]东南大学附属中大医院溧水分院泌尿外科,江苏南京211200 [2]江苏大学附属医院甲状腺乳腺外科,江苏镇江212001

出　　处：《现代医学》2020年第4期515-520,共6页Modern Medical Journal

摘　　要：目的:探讨利用乙醛酸盐诱导小鼠形成草酸钙结晶而导致的小鼠肾损伤中肾小管上皮细胞的早期变化。方法:选择乙醛酸盐作为诱导剂通过腹腔给小鼠注射而在肾部形成草酸钙结晶制作肾损伤动物模型;然后利用全自动生化分析仪测定血浆尿素氮和血肌酐水平的变化,利用HE染色法对肾脏组织进行染色观察肾脏结构的病理变化,钙盐染色法对肾脏组织染色观察钙盐沉积情况,并利用茜素红法对钙盐的沉积情况进行半定量测定,采用蛋白质印迹法观察肾小管上皮细胞上的标志蛋白E-钙黏蛋白(E-cadherin)、广谱细胞角蛋白(Pan-ck)和间质标志蛋白α-平滑肌肌动蛋白(α-SMA)、波形蛋白(Vimentin)的变化。结果:随着乙醛酸盐注射小鼠腹腔时间的延长,血肌酐和尿素氮均水平升高,差异具有统计学意义(P<0.05);HE染色结果显示可见肾皮质近曲小管处分布着相互连接的结晶,近曲肾小管的部分细胞疏松水肿,远曲小管发生扩张,且肾小管上皮细胞发生变形和肿胀,逐渐裸露出基底膜;与正常对照组比较,乙醛酸盐干预组的HE病理评分值逐渐升高,差异具有统计学意义(P<0.05);钙盐染色结果显示近端小管腔内的草酸钙结晶沉积量逐渐增加,差异具有统计学意义(P<0.05);蛋白印迹结果显示E-cadherin及Pan-ck的蛋白表达呈逐渐减少的现象,而间质α-SMA及Vimentin的蛋白表达量逐渐上升。结论:利用乙醛酸盐对小鼠诱导使其肾脏形成草酸钙结晶所致的肾损伤模型中,肾小管上皮细胞标志蛋白表达量减少,而间质标志蛋白表达量增加,发生了肾间质纤维化过程,从而为研究肾损伤的发生机制提供理论基础。Objective:To investigate the changes of renal tubular epithelial cells of injury kidney mode induced by crystallization of calcium oxalate.Methods:Animal model of calcium oxalate crystal was established by intraperitoneal injection of glyoxylate in mice.The changes of plasma urea nitrogen and serum creatinine were measured by automatic biochemical analyzer.Pathological changes of renal structure were observed by HE staining.The change of the expression of E-cadherin,Pan-cytokeritin(Pan-ck)and interstitial marker proteinα-smooth muscle actin(α-SMA),vimentin on renal tubular epithelial cells were observed by Western blot.Results:With the continuous intraperitoneal injection of glyoxylate time prolonged,the serum levels of creatinine and urea nitrogen were significantly higher(P<0.05),and there was statistically significant difference.HE staining showed that the proximal tubules of the renal cortex were distributed with interconnected crystals.Some of the cells of the proximal tubule were loose and edema,and the distal tubules were expanded and the renal tubular epithelial cells were deformed and swollen,and the basement membrane was gradually exposed.Compared with the normal control group,the histopathological grade of the other glyoxylic acid group was gradually increased,and the difference was statistically significant(P<0.05).Calcium salt staining showed that the calcium oxalate crystals deposited in the proximal tubule(P<0.05).The expression of E-cadherin and Pan-ck in renal tubule epithelium was decreased by western-blot,while the expression of interstitial marker proteinα-SMA and Vimentin was gradually increase.Conclusion:Glyoxylate-induced calcium oxalate crystals in mice resulted in decreased expression of renal tubular epithelial cells in mouse renal injury model,and the expression of interstitial marker protein increased,which starts the process of fibrosis.So as to provide a theoretical basis for the study of the mechanism of renal injury.

关 键 词：乙醛酸盐 草酸钙结晶 小鼠 肾损伤 上皮细胞 

分 类 号：R692[医药卫生—泌尿科学]