迷迭香酸对大鼠肾草酸钙结石形成的影响
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摘要
目的研究迷迭香酸对大鼠肾草酸钙结石形成的影响,探讨迷迭香酸的药理作用机制。
方法采用1%乙二醇和2%氯化铵ig造成大鼠草酸钙结石模型,应用迷迭香酸不同剂量作为干预因素,肾石通颗粒剂作为阳性对照药。采用生化全自动分析仪检测各组大鼠成石不同时间的尿生化及应用高效液相色谱法(HPLC)测定大鼠尿草酸、尿枸橼酸的含量,造模28d结束后生化仪检测血生化。解剖取双肾,HPLC法测定肾组织草酸、枸橼酸的含量。原子吸收光谱测定肾钙含量。HE染色观察肾组织病理变化、Von-Kossa’s染色观察肾草酸钙结晶形态分布。应用免疫组化方法观察间α胰蛋白酶抑制物(IαI)在肾组织中的表达情况。
结果迷迭香酸中剂量(0.05g·Kg-1)与高剂量(0.1g·Kg-1)组大鼠的24小时尿中肌酐、尿酸、草酸、尿Ca2+含量,血BUN、Cr、Ca2+含量,肾组织Ca2+含量,与结石模型组比较有显著差异(P<0.05);而迷迭香酸低剂量组与结石模型组比较无显著差异(P>0.05)。迷迭香酸中剂量与高剂量组血清BUN、Cr、Ca2+含量比较也有显著差异(P<0.05),迷迭香酸中、高剂量组肾组织病理变化、草酸钙结晶情况及肾小管上皮细胞IαI的表达对比结石模型组有明显不同。
结论迷迭香酸可以抑制大鼠肾草酸钙结石的形成,并有一定的量–效关系。
Objective To investigate rosmarinic acid's impact on calcium oxalate renal stone formation in rats and study the pharmacological actions and mechanisms of rosmarinic acid too.
Methods The model rats with renal calcium oxalate stone formation were induced by ig 1% ethylene glycol[V/V] and 2% ammonium chloride[W/V] for 28 days, and application of different dose of rosmarinic acid as the interference factor, renal stone kernels as positive control drug. Use automatic biochemical analyzer to detect the urinalysis biochemistry of rats in each group in different periods and High Performance Liquid Chromatography(HPLC) to determine the content of ethanedioicacid and citric acid in urine of rats. Examine the blood biochemistry after 28d. Dissect the rats and study their kidney, use HPLC to determinethe content of ethanedioic acid and citric acid in the kidney tissue. Use atomic absorption spectroscopy to determine the content of calcium in kidney. Hematoxylin-eosin staining to observe the renal pathological changes and Von-Kossa’s staining to observe renal calcium oxalate crystal morphology distribution. Use immunohistochemical methods to observe the expression ofinter-trypsin inhibitor (IαI) in kidney tissues.
Results Compare the creatinine, uric acid, oxalic acid, Ca2+ content inurine; BUN, Cr, Ca2+ content in serum, Ca2+ content in kidney tissue of both rosmarinic acid middle-dose(0.05g·Kg-1) and high-dose group(0.1g·Kg-1) to the renal stone group, there are significant difference (P <0.05); rosmarinic acid low-dose group and the renal stone model group showed no significant difference (P> 0.05). Rosmarinic acid middle-dose and high-dose group had significant difference (P <0.05) in BUN, Cr, Ca2+ content of serum, and they both had difference in renal pathological changes, calcium oxalate crystal morphology and the expression of IαI in kidney tubules epithelial cell compared to renal stone group.
Conclusion Rosmarinic acid can inhibit renal calcium oxalate stone formation in rats, and that’s a dose-response relationship.
方法采用1%乙二醇和2%氯化铵ig造成大鼠草酸钙结石模型,应用迷迭香酸不同剂量作为干预因素,肾石通颗粒剂作为阳性对照药。采用生化全自动分析仪检测各组大鼠成石不同时间的尿生化及应用高效液相色谱法(HPLC)测定大鼠尿草酸、尿枸橼酸的含量,造模28d结束后生化仪检测血生化。解剖取双肾,HPLC法测定肾组织草酸、枸橼酸的含量。原子吸收光谱测定肾钙含量。HE染色观察肾组织病理变化、Von-Kossa’s染色观察肾草酸钙结晶形态分布。应用免疫组化方法观察间α胰蛋白酶抑制物(IαI)在肾组织中的表达情况。
结果迷迭香酸中剂量(0.05g·Kg-1)与高剂量(0.1g·Kg-1)组大鼠的24小时尿中肌酐、尿酸、草酸、尿Ca2+含量,血BUN、Cr、Ca2+含量,肾组织Ca2+含量,与结石模型组比较有显著差异(P<0.05);而迷迭香酸低剂量组与结石模型组比较无显著差异(P>0.05)。迷迭香酸中剂量与高剂量组血清BUN、Cr、Ca2+含量比较也有显著差异(P<0.05),迷迭香酸中、高剂量组肾组织病理变化、草酸钙结晶情况及肾小管上皮细胞IαI的表达对比结石模型组有明显不同。
结论迷迭香酸可以抑制大鼠肾草酸钙结石的形成,并有一定的量–效关系。
Objective To investigate rosmarinic acid's impact on calcium oxalate renal stone formation in rats and study the pharmacological actions and mechanisms of rosmarinic acid too.
Methods The model rats with renal calcium oxalate stone formation were induced by ig 1% ethylene glycol[V/V] and 2% ammonium chloride[W/V] for 28 days, and application of different dose of rosmarinic acid as the interference factor, renal stone kernels as positive control drug. Use automatic biochemical analyzer to detect the urinalysis biochemistry of rats in each group in different periods and High Performance Liquid Chromatography(HPLC) to determine the content of ethanedioicacid and citric acid in urine of rats. Examine the blood biochemistry after 28d. Dissect the rats and study their kidney, use HPLC to determinethe content of ethanedioic acid and citric acid in the kidney tissue. Use atomic absorption spectroscopy to determine the content of calcium in kidney. Hematoxylin-eosin staining to observe the renal pathological changes and Von-Kossa’s staining to observe renal calcium oxalate crystal morphology distribution. Use immunohistochemical methods to observe the expression ofinter-trypsin inhibitor (IαI) in kidney tissues.
Results Compare the creatinine, uric acid, oxalic acid, Ca2+ content inurine; BUN, Cr, Ca2+ content in serum, Ca2+ content in kidney tissue of both rosmarinic acid middle-dose(0.05g·Kg-1) and high-dose group(0.1g·Kg-1) to the renal stone group, there are significant difference (P <0.05); rosmarinic acid low-dose group and the renal stone model group showed no significant difference (P> 0.05). Rosmarinic acid middle-dose and high-dose group had significant difference (P <0.05) in BUN, Cr, Ca2+ content of serum, and they both had difference in renal pathological changes, calcium oxalate crystal morphology and the expression of IαI in kidney tubules epithelial cell compared to renal stone group.
Conclusion Rosmarinic acid can inhibit renal calcium oxalate stone formation in rats, and that’s a dose-response relationship.
引文
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[6]李林,黄松明,赵三龙,等.迷迭香酸对大鼠抗Thy L膜增生性肾小球肾炎模型氧化刺激的影响.实用儿科杂志,2007,22(15):1170-1180
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