腺嘌呤灌胃联合高磷饮食诱导CKD-MBD大鼠模型的探讨  

The Discussion about the Method of Adenine Gavage Integrating High Phosphorus Dietary in Model Preparation of Chronic Kidney Disease-Mineral and Bone Disorder in Rats

作　　者：郭星云 杨荣禄 任秋月 袁丽莎 石凯峰[1] 孟祥飞[1] 刘世巍[1] 李同侠[1] 张宁[1] 柳诗意[1] GUO Xingyun;YANG Ronglu;REN Qiuyue(Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing,100102)

机构地区：[1]中国中医科学院望京医院,北京100102 [2]北京中医药大学,北京100029 [3]中国科学技术大学附属第一医院安徽省立医院,合肥230036

出　　处：《中国中西医结合肾病杂志》2024年第2期111-115,I0001,I0002,共7页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金青年项目(No.81904156);中央级公益性科研院所基本科研业务费专项资金资助项目(No.ZZ14-YQ-021)。

摘　　要：目的:观察腺嘌呤灌胃联合高磷饮食方法诱导的慢性肾脏病矿物质和骨代谢异常(CKD-MBD)大鼠模型。方法:34只SD大鼠随机分为对照组及模型组,每组17只。实验第1~4周,每日予250 mg/kg腺嘌呤混悬液灌胃及含1.2%高磷饲料喂养,第5~8周腺嘌呤灌胃改为隔日。对照组和模型组灌胃等体积高纯水。共给药8周。8周后杀检取材,苏木精-伊红(HE)、过碘酸雪夫(PAS)染色、马松(Masson)染色法观察两组大鼠肾脏病理,茜素红、HE染色观察主动脉钙化结节形成情况;HE染色观察胫骨组织形态,显微CT(Micro-CT)与CT-AN软件对股骨进行三维分析,检测皮质骨骨形态计量学参数;全自动生化仪测定血钙(Ca)、血磷(P)、血肌酐(Scr)、血尿素氮(BUN)、血清全段甲状旁腺激素(iPTH)水平。结果:与对照组比较,模型组大鼠的Scr、BUN、P和iPTH水平均显著升高(P<0.01),Ca2+水平明显降低(P<0.01);模型组肾小球结构紊乱,肾小球基底膜增厚,肾小管扩张,管腔内有积水、炎性细胞的浸润和棕褐色结晶沉积,肾脏血管明显减少,肾脏间质纤维化,有炎性细胞浸润;模型组大鼠主动脉中膜明显的呈连续性线性分布的钙化结节,局部血管破裂;模型组大鼠α-平滑肌肌动蛋白(α-SMA)蛋白平均光密度值表达明显降低,骨形态蛋白-2(BMP-2)和Runt相关转录因子2(Runx2)蛋白平均光密度值表达升高,差异有统计学意义(P<0.01);模型组大鼠胫骨近端干骺端的小梁骨网络之间的断开和分离增加,骨小梁表面的成骨细胞和破骨细胞数量增加,骨小梁周围出现大量间质纤维化,模型组皮质骨组织密度显著降低(P<0.01)、皮质孔隙度显著增加(P<0.01),皮质骨体积分数差异无统计学意义(P>0.05)。结论:腺嘌呤灌胃联合高磷饮食是制备CKD-MBD大鼠模型较为快速、可行、准确的方法,值得进一步推广。Objective:To observe the adenine gavage combined with high phosphorus diet method induced chronic kidney disease-mineral and bone disorder(CKD-MBD)rat model.Methods:34 SD rats were randomly divided into control group and model group,17 rats in each group.The rats were fed with 250 mg/kg adenine suspension by gavage and 1.2%high phosphorus diet daily from the 1st to the 4th week of the experiment,and the adenine gavage was changed to once per two days from the 5th to the 8th week.The control and model groups were given equal volumes of high purity water by gavage for 8 weeks.After 8 weeks,hematoxylin-eosin(HE),periodic acid Schiff(PAS)staining and Masson staining were used to observe the renal pathology of the rats in both groups.The morphometric parameters of cortical bone morphology were measured;the levels of blood calcium(Ca),blood phosphor⁃us(P),blood creatinine(Scr),blood urea nitrogen(BUN)and serum whole segment parathyroid hormone(iPTH)were determined by fully automated biochemical instrument.Results:Compared with the control group,the levels of Scr,BUN,P and iPTH were sig⁃nificantly higher(P<0.01)and Ca levels were significantly lower(P<0.01)in the model group;the glomerular structure of the model group was disturbed,with thickened glomerular basement membrane,dilated tubules,fluid accumulation,infiltration of inflam⁃matory cells and brown crystalline deposits in the tubular lumen,significantly reduced renal blood vessels,interstitial fibrosis and in⁃flammatory cell infiltration;the model group rats had obvious calcified nodules in the aortic mesothelium with continuous linear distri⁃bution and local vascular rupture;the mean optical density value ofα-smooth muscle actin(α-SMA)protein expression was signif⁃icantly decreased,and the mean optical density value of bone morphogenetic protein-2(BMP-2)and Runt-related transcription factor 2(Runx2)protein expression was increased in the model group rats,and the difference was statistically(P<0.01);increased disconnection and separation between trab

关 键 词：慢性肾脏病矿物质和骨代谢异常(CKD-MBD) 血管钙化 高转运型骨病 腺嘌呤 高磷饮食 

分 类 号：R-332[医药卫生] R692