采用黄嘌呤及氧嗪酸钾联合腹腔注射的方法建立符合人体高尿酸血症发病机制的稳定动物模型,比较二妙丸水提物苍术和黄柏比例分别为1:1,1:2和2:1的水提物(含生药360 mg·mL-1)及二妙丸 (360 mg·mL-1) 分别连续灌胃给予大鼠两周后,评价药物对高尿酸血症的治疗和保护作用.利用紫外可见分光光度法检测了血清及肝脏中黄嘌呤氧化酶(XOD)的活性变化.在570 nm处进行测定,结果显示,模型组与各组相比,血清中XOD活性显著升高(p<0.01),肝脏中XOD活性无显著差异(p>0.05),但各给药组与模型组相比血清和肝脏中XOD活性均显著降低(p<0.01),其中苍术和黄柏比例为1:2的组方疗效最为显著.对肾脏石蜡切片应用苏木精-伊红染色法(HE染色)及马松三色染色法(Masson染色)进行染色,光镜下观察模型组及各给药组肾小球的形态变化及肾小管间质的纤维化程度.结果表明,模型组肾小球萎缩、血管袢分辨不清、出现一定程度的炎细胞浸润、肾间质纤维化等现象.各给药组与模型组比较,肾小球萎缩、炎细胞浸润、纤维组织生成等均有所改善.纤维化面积结果显示,模型组与各组相比纤维化程度显著升高(p<0.01或p<0.05),但与二妙丸组的差异性低于其类方给药组(p<0.05),其中苍术和黄柏比例为1:2的组方疗效最显著.本实验以肾损伤和黄嘌呤氧化酶活性两个方面为考察指标,对高尿酸血症的治疗和保护作用进行评价,为后续二妙丸类方药物的开发提供科学依据。

A stable mouse model of hyperuricemia was established by intraperitoneal injection of xanthine and oxonate,comparing the water extracts (containing crude drug 360 mg·mL-1) of Ermiao pill categorized formula (The ratio of atractylodes lancea to cortex phellodendri was 1:1,1:2 and 2:1,respectively) and Ermiao pill (360 mg·mL-1) administered to different groups of animals continuously for two weeks and assessing the protection or treatment of drug on hyperuricemia.The xanthine oxidase (XOD) activities in serum and liver were detected by ultraviolet-visible spectroscopy at 570 nm wavelength,The results showed that as compared with each group,the XOD activity of the model group was significantly increased in serum (p<0.01),XOD activity showed no significant difference in liver (p>0.05),but compared with the model group,the XOD activity of each treatment group was significantly lower in serum and liver (p<0.01),especially for the group treated with Ermiao pill categorized formula with the ratio of Atractylodes lancea to Cortex Phellodendri being 1:2.The morphological changes of glomerular and tubular interstitial fibrosis were measured by Hematoxylin-eosin staining (HE staining) and Masson trichrome staining (Masson staining)for kidney paraffin sections.The results showed that the glomerular atrophy,vascular loops confusion,a certain degree of inflammatory cell infiltration,interstitial fibrosis and other phenomena appeared in the model group.Compared to model group,these pathological phenomena of the treatment groups were significantly improved.The area showed that compared with each group,the fibrosis of the model group was significantly increased (p<0.01 or p<0.05),but compared with Ermiao pill categorized formula,the differences for the group of Ermiao pill was lower (p<0.01),especially for the group treated with Ermiao pill categorized formula with the ratio of Atractylodes lancea to Cortex Phellodendri being 1:2.In this experiment,the damage of kidney and XOD activity serve as the index to evaluate the protection or treatment of drug on hyperuricemia,providing a scientific basis for the development of Ermiao pill categorized formula.