穿山龙总皂苷对痛风性关节炎大鼠尿液中生物标志物的影响

Effect of total saponins from Rhizoma Dioscorea nipponica on biomarkers in urine of gouty arthritis rats

关键词:  穿山龙总皂苷  痛风性关节炎  鸟苷  肌酐  尿酸  高效液相色谱法

Key words:  total saponins from Dioscorea nipponica Makino  gouty arthritis  guanosine  creatinine  uric acid  HPLC

摘要:

[目的] 应用高效液相-紫外检测法（HPLC）和紫外分光光度法测定穿山龙总皂苷（RDN）对痛风性关节炎大鼠尿液中鸟苷、尿酸、肌酐含量的影响。[方法] 60只雄性Wistar大鼠随机分为6组，分别为正常组、模型组、穿山龙总皂苷高剂量组（160 mg/kg）、中剂量组（80 mg/kg）、低剂量组（40 mg/kg），和阳性药秋水仙碱组（0.3 mg/kg）.连续灌胃穿山龙总皂苷7 d.第3天灌胃1 h后，膝关节上方髌上韧带进针，注射0.2 mL浓度为25 g/L尿酸钠晶体混悬液到关节腔，造成实验性痛风性关节炎模型。采用苏木精-伊红（HE）和伊红染色观察关节滑膜组织病理学变化。鸟苷的测定采用 Diamonsil C₁₈（5 μm，250 mm×4.6 mm）色谱柱，以甲醇为流动相A，0.03 mol/L醋酸铵缓冲液（pH=4.65）作为流动相B，梯度洗脱，柱温25 ℃，流速为1 mL/min，检测波长254 nm；尿酸和肌酐的含量测定采紫外分光光度法检测。[结果] 痛风性关节炎模型组尿液中鸟苷和肌酐含量显着降低；尿酸含量显着升高。穿山龙给药组与模型组相比，鸟苷、尿酸和肌酐含量显着升高。[结论] 鸟苷、肌酐与尿酸均为痛风性关节炎发生发展的重要生物标志物，穿山龙总皂苷可通过调节其含量变化对痛风性关节炎起潜在治疗作用。

Abstract:

[Objective] To explore the effect of total saponin of Rhizoma Dioscorea nipponica (RDN) on contents of guanosine, uric acid and creatinine in urine of gouty arthritis rats by high performance liquid chromatography (HPLC) with UV detection and UV spectrophotometric. [Methods] The 60 Wistar rats were randomly divided into six groups. They were normal group, model group, total saponins groups of high (160 mg/kg) ,middle (80 mg/kg) and low(40 mg/kg) doses and colchicine group. Total saponins of RDN were given for 7 successive days. An hour after total saponins of RDN were given at the third day, 0.2 mL, 25 mg/mL MSU suspension was injected into articular cavity through anadesma of kneecap in the knee-joint to induce model of gouty arthritis. HE and eosin dyeing were used to observe the histopathological change. A Diamonsil C₁₈ column(5 μm, 250 mm×4.6 mm) was used for the analysis at 25℃, The separation was carried out with the mobile phase consisting of methanol-ammonium acetate(0.03 mol/L) at a flow rate of 1mL/min. The eluates were monitored by the programmed wavelength at 254 nm for guanosine; The contents of uric acid and creatinine in urine were detected by UV spectrophotometric. [Results] In the model group, the levels of guanosine and creatinine were significantly decreased and the content of uric acid was significantly increased. Compared with the model group, guanosine, uric acid and creatinine contents increased significantly in total saponins groups. [Conclusion] Guanosine, creatinine and uric acid were important biological markers in the occurrence and development of gouty arthritis. Total saponin of RDN have potential effects on the treatment of gouty arthritis by regulating the content change of these biomarkers.