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目的:测定应用防石合剂前后草酸钙模型大鼠肾组织OPN表达和尿中尿钙及尿尿酸的浓度变化,探讨防石合剂预防肾结石复发的作用机制。方法:用乙二醇+氯化胺法构建草酸钙肾结石大鼠模型,同时给予防石合剂1.50g生药材/1.5mL/100g、3g生药材/1.5mL/100g、6g生药材/1.5mL/100g,各组均给药7周。实验结束后检测各组大鼠24h尿量、尿PH、尿尿酸、尿钙,免疫组化法检测OPN蛋白表达,实时荧光定量PCR技术检测大鼠肾骨桥蛋白mRNA的表达。结果:模型组大鼠肾OPN mRNA及蛋白的表达明显增加,为对照组的2倍,防石合剂各剂量组均能进一步增加大鼠肾组织OPN mRNA及蛋白的表达,为对照组的3.5倍;且防石合剂各剂量组能显著降低大鼠尿中尿钙和尿尿酸含量,增加排尿量。结论:防石合剂可能通过增强草酸钙模型大鼠肾组织OPN的表达,从而抑制结石形成。
OBJECTIVE: To determine the changes of OPN expression in urine and urinary calcium and urine uric acid concentrations in kidney of calcium oxalate model rats before and after application of anti-stone mixture, and to explore the mechanism of anti-stone mixture in preventing relapse of kidney stones. Methods: Calcium oxalate nephrolithiasis rat model was constructed by ethylene glycol + amine chloride method, and 1.50g antibacterial agent / 1.5mL / 100g, 3g crude drug / 1.5mL / 100g, 1.6g crude drug / 1.5mL / 100g, each group were administered for 7 weeks. At the end of the experiment, 24h urine, urine PH, urine uric acid and urinary calcium were detected in all groups. The expression of OPN protein was detected by immunohistochemistry and the mRNA expression of osteopontin was detected by real-time fluorescence quantitative PCR. Results: The expression of OPN mRNA and protein in kidney increased significantly in model group, which was twice as that in control group. Each dose of anti-stone agent could further increase the expression of OPN mRNA and protein in rat renal tissue, which was 3.5 times that of control group ; And anti-stone mixture dose group can significantly reduce urinary urinary calcium and urinary uric acid content, increase urinary output. Conclusion: The stone inhibitor may inhibit the formation of calculus by enhancing the expression of OPN in renal tissue of calcium oxalate model rats.