补脾益肾中药对阿霉素诱导大鼠慢性肾损害肾组织p21和TGF-β1的影响

Effect of reinforcing spleen and kidney on the p21 and TGF-β1 in renal tissue of adriamycin-induced CKD in rats

目的 研究补脾益肾中药对阿霉素诱导大鼠慢性肾损害肾脏病变的影响,探讨其对肾间质纤维化和肾小球硬化的延缓作用.方法 健康雄性SD大鼠36只,随机分为模型组和空白对照组；模型建立后,模型组大鼠又随机分为模型对照组、苯那普利组和中药低、中、高剂量组;观察造模后第14天、第28天、第42天各组大鼠的一般情况、生化指标,第42天实验结束时处死大鼠取肾,用免疫组织化学染色方法测定各组大鼠肾组织中p21、TGF-β1表达的变化.结果 阿霉素诱导大鼠慢性肾损害的大鼠7天后出现蛋白尿,14d尿蛋白排泄达到高峰；中药治疗组、苯那普利组均明显降低阿霉素肾病大鼠的24 h尿蛋白量（P＜0.05）,其中中药高剂量组疗效更显著；肾组织病理切片示模型组大鼠肾组织中p21 （518886.35±6810.89）和TGF-β1 （222012.95±50484.73）表达明显增加（P＜0.05）;中药高剂量治疗组p21 （288627.66±97021.65）和TGF-β1（98405.14±19216.89）表达明显低于模型对照组（P＜0.05）.结论 补脾益肾法中药复方制剂可能通过调节肾组织中p21和TGF-β1的表达以减少蛋白尿从而保护肾脏,延缓肾脏病的进展.

Objective To investigate the effect of reinforcing spleen and kidney method on adriamycin-induced CKD in rats and to explore its possible mechanism.Methods Totally 36 Sprague-Dawley rats were randomly divided into a Adriamycin-induced model group and a control group.The model group was further divided into five groups：the Adriamycin-induced model control group,bennazepril-treated group,and TCM treated low,moderate,and high dose groups.The level of serum creatinine,urea nitrogen,24hours urine protein and urine creatinine were measured at 14,28,42 days after establishing the model rats.And the protein expression of transforming growth factor-β1 （TGF-β1） and cyclin-dependent kinase inhibitor p21cip1 （p21）were detected by immunohistochemistry.Results The proteinuria was observed on the seventh day after injection of adriamycin in adriamycin nephropathy model group,and reached summit on the fourteenth day.Both TCM treated groups and benazepril group reduced the level of urine protein within 24 hours （P〈0.05）,the reduction was most remarkable in the TCM high dose group.The expression of p21 and TGF-β1 （p21 288627.66±97021.65,TGF-β1 98405.14± 19216.89） in kidney increased in the model groups,while the TCM treated high dose group （p21 518886.35±6810.89,TGF-β1 222012.95± 50484.73） was significantly lower than the model control group （P〈 0.05）.Conclusion Reinforcing spleen and kidney method could decrease the level of urine protein within 24 hours by regulating the expression of p21 and TGF-β1,so thus to protect renal function and delay progress of kidney disease.