【摘要】 目的建立一种醛糖还原酶抑制剂筛选的细胞模型,避免中药对传统离体筛选方法的干扰,并对筛选所得药物改善糖尿病肾病(DN)的效应和机制进行研究。方法选用肾小球系膜细胞,确定最佳孵育条件,采用酶促微量反应法测定细胞内山梨醇含量,建立细胞-酶法-多元醇通路活性评价体系,以此方法筛选所得的具有明显抑制醛糖还原酶活性作用的葛根、知母组成葛知汤,用DN大鼠模型评价葛知汤改善DN的效应。结果细胞模型的最佳反应条件为含30mmol·L-1葡萄糖培养基孵育4h。葛根和知母能明显抑制细胞模型和动物模型的多元醇通路激活,也能纠正模型动物的氧化还原失衡,改善肾脏功能,减轻肾小球系膜基质增生。结论该细胞模型能够有效排除中药对酶法测定醛糖还原酶活性的干扰,模拟体内的反应情况,筛选所得的中药醛糖还原酶抑制剂能有效改善DN模型的肾脏功能。更多还原

【Abstract】 Objective To develop a cell model suitable for herbal aldose reductase inhibitor screening and to verify the ameliorative effects of the inhibitor on diabetic nephropathy rat model.Methods The mesangial cell was used in the herbal aldose reductase inhibitor screening system and the optimal incubating conditions were investigated.Enzymatic microreaction method was used for the assay of intracellular D-glucitol content,and a cell-enzyme-polyol pathway activities evaluation system was established.By this system,ameliorative effects and possible mechanisms of the herbal aldose reductase inhibitor from Gezhi Decoction were investigated on diabetic nephropathy rats.Results The optimal incubating conditions were incubating the cells for 4 hours with medium containing 30 mmol·L-1 glucose.Radix Puerariae and Rhizoma Anemarrhenae inhibited the activity of aldose reductase significantly,corrected the imbalance of oxidase and reductase in rat model,improved renal function and reduced hyperplasia of mesangial matrix.Conclusion The developed cell model can effectively eliminate the disturbance of herbs on the traditional aldose recuctase inhibitor screening system and has mimic in-vivo diabetic status preferably.Radix Puerariae and Rhizoma Anemarrhenae screened with the cell model can ameliorate diabetic nephropathy in the rat model.更多还原