贵州医科大学学报

2019, v.44;No.228(09) 1049-1053+1058

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炎症状态下终末期肾病大鼠肾脏Klotho蛋白表达及血管钙化的关系
Expression of Klotho Protein and Vascular Calcification in Kidney of Rats with End-stage Renal Disease under Inflammatory State

郭莉;李建东;郭珊珊;陈航;龚幼兰;张亚璞;
GUO Li;LI Jiandong;GUO Shanshan;CHEN Hang;GONG Youlan;ZHANG Yapu;Department of Kidney Medicine, Affiliated Hospital of Hebei University;

摘要(Abstract):

目的:探讨炎症状态下终末期肾病(ESRD)大鼠Klotho蛋白与血管钙化的关系。方法:将大鼠随机分为对照组(灌胃生理盐水)、ESRD组(250 mg/kg灌胃腺嘌呤)、ESRD合并炎症组(250 mg/kg灌胃腺嘌呤,隔日以1.0 g/kg皮下注射10%酪蛋白),于灌胃10周时,采用化学比色法检测血清钙(Ca)、磷(P)、尿素氮(BUN)及肌酐(SCr)水平,采用ELISA法检测血清甲状旁腺激素(iPTH)水平,采用ELISA法测定大鼠肾脏组织Klotho蛋白水平、丙二醛(MDA)、过氧化氢酶(CAT)含量;采用免疫组化技术检测肾脏组织Klotho蛋白表达,HE染色观察肾组织形态学改变,茜素红染色观察肾组织钙盐沉积。结果:灌胃10周时,各组大鼠血清P、SCr、BUN、iPTH及肾组织中MDA比较,ESRD组、ESRD合并炎症组均高于对照组(P<0.01);各组大鼠血清Ca浓度及肾脏组织中CAT含量比较,ESRD组、ESRD合并炎症组低于对照组(P<0.05),ESRD组血清Ca浓度低于ESRD合并炎症组(P<0.05);Klotho蛋白表达主要集中在肾小管,ESRD组、ESRD合并炎症组肾脏组织Klotho蛋白表达水平显著低于对照组(P<0.05);HE及茜素红染色结果显示,ESRD合并炎症组出现肾小球萎缩及硬化,肾内小动脉出现较多片状钙盐沉积,肾内小动脉存在明显钙化现象。结论:炎症可加重ESRD大鼠血管钙化,其机制可能与降低Klotho蛋白表达有关。
Objective: To investigate the relationship between Klotho protein and vascular calcification in end-stage kidney disease(ESRD) rats with inflammation. Methods: Rats were randomly divided into control group(gastric saline), ESRD group(250 mg/kg gastric adenine) and ESRD complicated with inflammation group(250 mg/kg gastric adenine, subcutaneous injection of 10% casein at 1.0 g/kg every other day). The levels of serum calcium(Ca), phosphorus(P) urea nitrogen(BUN) and creatine(SCr) were measured by chemical colorimetric method. Serum parathyroid hormone(iPTH) level was measured by ELISA method. The level of Klotho protein, malondialdehyde(MDA) and the content of catalase(CAT) in rat kidney were measured by ELISA method. The expression of Klotho protein in renal tissue was detected by immunohistochemistry. The morphological changes of renal tissue were observed by HE staining, and the calcium deposition in renal tissue was observed by alizalin red staining. Results: At 10 weeks after intragastric administration, the serum P, SCr, BUN, iPTH and MDA in renal tissue of rats in each group were higher than those in ESRD group and ESRD complicated with inflammation group(P<0.01).The concentration of serum Ca and the content of CAT in kidney tissue of rats in each group were lower than those in control group in ESRD group and ESRD complicated inflammation group(P<0.05). The concentration of serum Ca in ESRD group was lower than that in ESRD complicated with inflammation group(P<0.05). The expression of Klotho protein was mainly concentrated in renal tubules. The expression of Klotho protein in renal tissue of ESRD group and ESRD complicated with inflammation group was significantly lower than that of control group(P<0.05). The results of HE and alizalin red staining showed that in ESRD complicated with inflammation, there were atrophy and sclerosis of glomeruli, more flake calcium deposits were found in renal arterioles, and obvious calcification was found in renal arterioles. Conclusion: Inflammation can aggravate vascular calcification in ESRD rats, and its mechanism is related to the decrease of Klotho protein expression.

关键词(KeyWords): Klotho蛋白;终末期肾病;血管钙化;钙磷代谢;炎症
klotho protein;end-stage renal disease;vascular calcification;calcium and phosphorus metabolism;inflammation

Abstract:

Keywords:

基金项目(Foundation): 河北省科技计划项目(162777230)

作者(Author): 郭莉;李建东;郭珊珊;陈航;龚幼兰;张亚璞;
GUO Li;LI Jiandong;GUO Shanshan;CHEN Hang;GONG Youlan;ZHANG Yapu;Department of Kidney Medicine, Affiliated Hospital of Hebei University;

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DOI: 10.19367/j.cnki.1000-2707.2019.09.012

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