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-现代医学

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

单位：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

分类号：R692

出版年·卷·期（页码）：2020·39·第四期（515-520）

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 protein α-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 protein α-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.

参考文献：

[1] 徐汉军,刘春林,单勇,等.组合式输尿管软镜钬激光碎石术治疗肾结石术后结石残留的原因分析[J].现代医学,2016,44(6):846-848.
[2] 李小琴,欧阳炎,陈磊.象山半岛渔民肾结石患病现状及危险因分析[J].浙江医学,2017,39(14):1207-1209.
[3] 何冉,丁鑫,崔绍琼.输尿管软镜在肾结石合并复杂尿路感染治疗中的疗效分析[J].中外医学研究,2018,16(14):146-148.
[4] 白福鼎,吴慧锋,文甲明,等.预先留置双J管在经皮肾镜取石术中应用的临床分析[J].中华医学杂志,2018,98(6):454-456.
[5] 罗怀景,林超禄,陈建德,等.输尿管软镜治疗肾结石的临床疗效及对肾功能的影响作用[J].中国现代医生,2017,55(29):47-49.
[6] 王翔,黎承杨,汪小明,等.电子输尿管软镜与纤维输尿管软镜治疗上尿路结石的疗效对比研究[J].临床泌尿外科杂志,2018,33(2):93-95.
[7] 付贤,王旭亮,李莉,等.超微经皮肾镜与输尿管软镜治疗肾结石疗效比较[J].浙江临床医学,2018,20(5):906-908.
[8] 夏生俊,浦金贤,张学锋,等.微通道经皮肾镜碎石术和输尿管软镜碎石术治疗直径10至20毫米肾结石肥胖患者的疗效比较[J].中国医师进修杂志,2018,41(11):994-998.
[9] 蔡华芳.草酸钙肾结石模型及其应用[J].中国中西医结合肾病杂志,2010,11(8):750-752.
[10] CHEN YH,LIU HP,CHEN HY,et al.Ethylene glycol induces calcium oxalate crystal desposition in Mailpghian tubules:a drosophila model for nephrolithiasis/ urolithiasia[J].Kidney Int,2011,80(4):369- 377.
[11] THAMILSELVAN S,BYER K J,HACKETT R L,et al.Free radical scawengers,catalase and superoxide dismutase provide protection from oxalate-associated injury to LLC-PKI and MDCK cell[J].J Urol,2000,164(1):224-229.
[12] LI Y H,YU S L,GAN X G,et al.Externalization of phosphatidylserine via multidrug resistance 1(MDR1)/P -glycoprotein in oxalate-treated renal epithelial cells:implications for calcium oxalate urolithiasis[J].Int Urol Nephrol,2016,48(2):175-181.
[13] KHAN A H,IMRAN S,TALATI J,et al.Fourier transform infrared spectroscopy for analysis of kidney stones[J].Investig Clin Urol,2018,59(1):32-37.
[14] SHOAR K,TURNEY B W,CLEVELAND R O.Tracking kidney stones in a homogeneous medium using a trilateration approach[J].Acoust Soc Am,2017,142(6):3715-3721.
[15] MAGHSOUDI R,ETEMADIAN M,SHADPOUR P,et al.Number of tracts orstone size:which influences outcome[J].Urol Int,2017,89(1):103-106.
[16] DONGRE A R,RAJALAKSHMI M,DESHMUKH P R,et al.Risk factors for kidney stones in rural puducherry:case-controlstudy[J].Clin Diagn Res,2017,11(9):LC01-LC05.
[17] YARIMOGLU S,POLAT S,BOZKURT I H,et al.Comparison of S.T.O.N.E and CROES nephrolithometry scoring systems for predicting stone-freestatus and complication rates after percutaneous nephrolithotomy:a single center study with 262 cases[J].Urolithiasis,2016,18(2):256-258.
[18] WANG R L,DING G,LIANG W,et al.Role of LOX-1 and ROS in oxidized low-density lipoprotein induced epithelial-mesenchymal transition of NRK52E[J].Lipids Health Dis,2010,19(9):120.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录