姜黄素对阿霉素肾病大鼠NADPH氧化酶和nephrin表达的影响

Effect of Curcumin on Expression of NAPDH Ofxidase and Nephrin in the Renal Tissue of Rats with Adriamycin-induced Nephrosis

目的通过观察姜黄素对阿霉素肾病大鼠肾组织中尼克酰胺腺嘌呤二核苷酸磷酸(NADPH),氧化酶与肾小球裂隙膜分子nephrin表达的影响,探讨姜黄素治疗肾病综合征的作用机制。方法将Sprague-Dawley大鼠50只随机分为5组(正常对照组、模型组、姜黄素低剂量干预组、姜黄素高剂量干预组和波尼松干预组)。除正常组大鼠外,各组大鼠尾静脉一次性注射阿霉素(5 mg/kg)造模。分别在建模后第28天收集24 h尿液,第29天处死所有大鼠,取肾脏组织,应用化学比色法检测肾组织氧化应激指标丙二醛(MDA)和超氧化物歧化酶(SOD),实时荧光定量聚合酶链式反应法和蛋白质印迹法检测肾组织NADPH氧化酶活性的关键亚基p47phox和neprhin mRNA与蛋白的表达变化。结果与正常对照组相比,模型组大鼠24 h尿蛋白量、肾组织MDA含量明显增加,肾组织p47phoxmRNA和蛋白表达明显增加,SOD活性明显下降;肾组织中nephrin mRNA和蛋白表达均明显减少。与模型组比较,姜黄素干预组尿蛋白、肾组织MDA明显减少,SOD活性明显增加。姜黄素干预组可显著下调p47phoxmRNA和蛋白表达;姜黄素干预组nephrin mRNA和蛋白表达均明显增加。结论姜黄素能减轻肾小球滤过屏障损伤,降低尿蛋白,抑制肾组织p47phox表达和增加nephrin表达可能是其作用机制之一。

Objective To explore the mechanisms of curcumin in treating nephrotic syndrome by observing the alter- ation of nicotiuamide adenine dinucleotide phosphate （NADPH） oxidase and nephrin in renal tissue of rats with adriamycin （ADR） -induced nephrosis. Methods Nephrosis was induced by single intravenous injection of ADR （5 mg/kg）. Fifty Sprague-Dawley rats were randomly divided into five groups： control group, nephrosis model group, nephrosis rats treated with low-dose and high-dose curcumin, and nephrosis rats treated with glucocortieoid. The 24-hour urine was collected 28 days after ADR injection to measure 24-hour urinary protein, and all rats were sacrificed 29 days after ADR injection. The levels of malondialdehyde （MDA） and the activeity of superoxide dismutase （ SOD ） in renal tissue were determined by chemiluminescence assay, and the mRNA and protein expression of NADPH oxidase p47phox and nephrin in renal tissue were determined by real time polymerase chain reaction and western blotting. Results The levels of urinary protein increased significantly in ADR-induced nephrosis group compared to control group ; the treatment of curcumin could reduced markedly urinary protein compared to ADR-induced neprhosis group. Compared with control group, ADR-indueed nephrosis group showed an elevation in MDA levels and a reduction in SOD activity in renal tissue. Treatment of curcumin markedly de-creased the levels of MDA and increased the activity of SOD compared to ADR-induced nephrosis group. Compared with control group, the mRNA and protein expression of p47phox in ADR-induced nephrosis groups were significantly increased. Treatment of curummin significantly decreased the expression of p47phox in renal tissue compared to ADR-induced nephrosis group. Compared with control group,the mRNA and protein expression of nephrin in ADR-induced nephrosis were signifi- cantly reduced. Treatment of curummin significantly increased the expression of nephrin compared to ADR-induced nephro- sis group. Conclusion Curcumin reduced excretion of urine protein in response to adriamycin via downregulation of p47phox expression and upregulation of nephrin expression. Thereby,curcumin may protect against ADR-induced nephrosis.