调整腺嘌呤饮食磷含量制作慢性肾病高磷血症的小鼠模型

Adjusting phosphorus content in adenine diet to establish mice model of chronic kidney disease with hyperphosphatemia

目的:通过调整腺嘌呤饮食中的磷水平,建立慢性肾脏病伴高磷血症的小鼠模型。方法:将雄性C57BL/6小鼠分为腺嘌呤饮食组(腺嘌呤0.2%,钙1.0%,磷0.6%)及其对照组(钙1.0%,磷0.6%)、腺嘌呤联合高磷饮食组(腺嘌呤0.2%,钙0.6%,磷1.0%)及其对照组(钙0.8%,磷0.6%),每组各7只。于造模前、造模4周记录体重,测血尿素氮(BUN)、钙(Ca)、磷(P)水平,4周后取肾脏组织,RT-PCR检测肾纤维化指标包括Ⅰ型胶原蛋白(CollagenⅠ)、纤连蛋白(fibronectin,FN)、纤溶酶原激活物抑制剂-1(plasminogen activator inhibitor-1,PAI-1)以及炎症指标肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)等的表达情况。结果:与相应的对照组比,腺嘌呤饮食组4周时体重降低,血BUN明显升高,为(41.15±4.59)mmol/L;Ca、P轻度升高,分别为(2.62±0.16)mmol/L、(2.22±0.26)mmol/L(P<0.05)。腺嘌呤联合高磷饮食组造模4周时血BUN[(14.68±3.57)mmol/L]轻度升高、P[(2.97±0.29)mmol/L]明显升高(P<0.05),血Ca水平无明显差异。RT-PCR显示腺嘌呤饮食组和腺漂呤联合高磷饮食组小鼠肾脏组织中CollagenⅠ、FN、PAI-1、TNF-α、IL-1β、ICAM-1表达水平均较相应对照组升高。结论:腺嘌呤联合高磷饮食饲喂C57BL/6小鼠4周即可建立高磷血症模型,同时伴有轻度肾功能下降、肾脏纤维化,是制备慢性肾脏病伴高磷血症小鼠模型的可行方法。

Objective：By adjusting the phosphorus content in adenine diet,we aimed to establish an effective chronic kidney disease mice model accompanied by hyperphosphatemia for further studies. Methods：Male C57 BL/6 mice were divided into four groups：simple adenine dietary：the normal group（1.0% calcium,0.6% phosphorus）,the simple adenine group（0.2% adenine,1.0%calcium,0.6% phosphorus）,adenine integrating high phosphorus dietary：the normal group（0.8% calcium,0.6% phosphorus）,the adenine integrating high phosphorus dietary group（0.2% adenine,0.6% calcium,1.0% phosphorus）（n=7）. The weight,the levels of blood urea nitrogen（BUN）,calcium（Ca）and phosphorus（P）in serum were measured at 0 and 4 weeks after the start of adenine diet.The gene expression of fibrosis markers collagenⅠ,fibronectin（FN）,plasminogen activator inhibitor-1（PAI-1）,inflammatory markers TNF-α,IL-1β and ICAM-1 in the kidney were detected by RT-PCR. Results：Compared to the control group,their bodyweights were decreased and the serum BUN level[（41.15 ± 4.59）mmol/L]was significantly increased at 4 weeks in the simple adenine group,while the levels of serum Ca[2.62 ± 0.16）mmol/L]and P[（2.22 ± 0.26）mmol/L]were slightly increased（P〈0.05）. In the adenine integrating high phosphorus dietary group,their weights were reduced and the level of serum P[（2.97 ± 0.29）mmol/L]showed significant increase,whereas the level of serum BUN[（14.68 ± 3.57）mmol/L]were slightly increased at 4 weeks（P〈0.05）. There was no significant difference in serum Ca levels. RT-PCR results showed that the expressions of collagenⅠ,FN,PAI-1,TNF-α,IL-1β and ICAM-1 were increased in two model groups. Conclusion：Adenine combined with high phosphorus diet feeding C57 BL/6 mice for 4 weeks can establish a hyperphosphatemia model,accompanied by mild renal dysfunction and renal fibrosis,which is an effective method to set up the mice with hyperphosphatemia.