肾性骨病方对5/6肾切除术后小鼠骨丢失影响的作用机制研究

Study on the Mechanism of Shenxinggubing Decoction on Bone Loss in Mice after 5/6 Nephrectomy

目的:比较肾性骨病方对不同阶段5/6肾切除小鼠骨丢失的保护作用机制。方法:3个月龄C57BL/6J小鼠150只,建立慢性肾衰小鼠模型,术后随机分为假手术组、模型组、罗盖全组、肾性骨病方组,分别在术后1周和4周时给予肾性骨病方以及罗盖全灌胃。各组给药8周后,取小鼠L5椎体、血清和肾脏组织,分别检测血清肌酐(CREA)、尿素氮(BUN),以评估肾脏功能;L5椎体进行阿尔辛兰/橙黄G染色、Micro-CT扫描加重建、免疫组化检测评估骨质情况。结果:和假手术组相比,模型组小鼠血清肌酐、血清尿素氮明显升高;和模型组相比,早期应用肾性骨病方干预后小鼠腰椎骨丢失情况较晚期各组得到明显改善;早期组小鼠L5腰椎Osterix表达明显升高而Cathepsin K表达明显降低;与之一致,TRAP染色显示早期应用肾性骨病方可以显著抑制破骨细胞活性,降低破骨细胞数量。结论:早期应用肾性骨病方可以有效改善5/6肾切除小鼠肾功能,缓解骨丢失。

Objective:To compare the protective mechanism of shenxinggubing decoction on bone loss in 5/6 nephrectomized mice at different stages. Methods:A total of 150 male C57 BL/6 J mice of 3-month-old were selected to establish the mouse model of chronic renal failure and randomly divided into sham group, model group, Shenxinggubing(SXGBF) group and Rocaltrol group. Shenxinggubing decoction and Rocaltrol were orally administrated at postoperative 1 week stage and 4 weeks stage respectively. After 8 weeks administration, L5 vertebrae, serum and kidney tissue of mice were taken to detect serum creatinine(CREA) and blood urea nitrogen(BUN). Arsine blue/orange G, Micro-CT scanning and reconstruction, and immunohistochemistry was used to evaluate bone condition in lumbar 5 vertebrae(L5). Results:Compared with sham group, serum CREA and BUN were significantly increased in model group. Compared with the model group, the bone loss of mics lumbar vertebra in the groups with early intervention of SXGBF decoction were significant improved than the late groups. In the early group, the expression of osterix in L5 lumbar vertebra was significantly increased and Cathepsin K was significantly decreased. In accordance with this, TRAP staining showed that early application of kidney osteopathy prescription could significantly inhibit the activity of osteoclasts and reduce the number of osteoclasts. Conclusion:The early application of SXGBF decoction effectively relieve renal insufficiency and bone loss in 5/6 nephrectomized mice.