补肾活血祛风方对阿霉素肾病大鼠肾组织NF-κB及IL-18水平的干预研究

Effect of Bushen Huoxue Qufeng Formula on levels of NF-κB and IL-18 in renal tissue of rats with adriamycin nephrosis

目的观察补肾活血祛风方对阿霉素肾病大鼠肾组织中核因子-κB(NF-κB)及白介素-18(IL-18)表达的影响,探讨补肾活血祛风方治疗肾病综合征的可能机制。方法60只SD雄性大鼠随机分为5组:空白组、模型组、泼尼松组、补肾活血祛风方组和补肾活血祛风方+泼尼松组。空白组和模型组每天给予蒸馏水灌胃;泼尼松组给予醋酸泼尼松片混悬液灌胃,剂量为6 mg/kg;补肾活血祛风方组给予补肾活血祛风方药液灌胃,剂量17.19 g/kg;补肾活血祛风方+泼尼松组上午以醋酸泼尼松片混悬液6 mg/kg灌胃,下午以补肾活血祛风方药液17.19 g/kg灌胃;连续灌胃4周。光镜和电镜观察肾组织病理结构变化;免疫组化法检测肾组织NF-κB-p65、IL-18的平均光密度;Real-time PCR法检测NF-κB-p65、IL-18的mRNA表达含量。结果与空白组比,模型组电镜下观察足细胞广泛融合,NF-κB-p65及IL-18的平均光密度上调(P<0.05),NF-κB-p65及IL-18的mRNA表达上调(P<0.05)。与模型组比,电镜下泼尼松组、补肾活血祛风方组、补肾活血祛风方+泼尼松组各组足细胞融合减轻,多呈节段性融合;泼尼松组、补肾活血祛风方组、补肾活血祛风方+泼尼松组NF-κB-p65、IL-18的平均光密度下调(P<0.05),NF-κB-p65、IL-18的mRNA表达下调(P<0.05)。电镜下补肾活血祛风方+泼尼松组足细胞融合程度最轻;补肾活血祛风方+泼尼松组IL-18的平均光密度较补肾活血祛风方组和泼尼松组下调(P<0.05)。结论泼尼松组、补肾活血祛风方组、补肾活血祛风方+泼尼松组可能通过下调NF-κB-p65、IL-18的mRNA及其蛋白表达,改善大鼠足细胞融合程度,从而缓解肾脏炎症反应、延缓疾病进展,且补肾活血祛风方+泼尼松组在疗效上最优。

Objective To observe the effect of Bushen Huoxue Qufeng(Kidney-tonifying Blood-invigorating and Wind-dispelling)(BHQ)Formula on the expression of nuclear factor-κB(NF-κB)and interleukin-18(IL-18)in the renal tissue of rats with adriamycin nephropathy,and explore the possible mechanism of BHQ Formula in treating nephrotic syndrome.Methods 60 SD male rats were randomly divided into 5 groups:blank group,model group,prednisone group,BHQ group and BHQ+prednisone group.The blank group and the model group were given distilled water by gavage every day;the prednisone group was given the prednisone acetate tablet suspension by gavage at a dose of 6 mg/kg;the BHQ group was given the BHQ decoction by gavage at a dose of 17.19 g/kg;the BHQ+prednisone group was given prednisone acetate tablet suspension 6 mg/kg by gavage in the morning,and BHQ decoction 17.19 g/kg in the afternoon.All groups were gavaged continuously for 4 weeks.The pathological changes of renal tissue were observed with light microscope and electron microscope;the average optical density of NF-κB-p65 and IL-18 in renal tissue was detected by immunohistochemical method;the mRNA expression of NF-κB-p65 and IL-18 was detected by Real-time PCR method.Results Compared with that of the blank group,in the model group the extensive fusion of podocytes was observed,the average optical density of NF-κB-p65 and IL-18 was up-regulated(P<0.05),and the mRNA expression of NF-κB-p65 and IL-18 was up-regulated under the electron microscope(P<0.05).Compared with that of the model group,in the prednisone group,BHQ group and BHQ+prednisone group,the podocyte fusion was reduced under electron microscope,with segmentary fusion being found in most groups,the mean optical density of NF-κB-p65 and IL-18 were down-regulated(P<0.05),and the mRNA expression of NF-κB-p65 and IL-18 was down-regulated(P<0.05).The degree of podocyte fusion was the least in the BHQ+prednisone group under electron microscope.The mean optical density of IL-18 in the BHQ+prednisone group was lower than that in the BHQ group and prednisone group respectively(P<0.05).Conclusion Prednisone group,BHQ group and BHQ+prednisone group may reduce the expression of NF-κB-p65,IL-18 mRNA and their protein to improve the degree of podocyte fusion,thereby alleviating renal inflammation and delaying disease progression,and the BHQ+prednisone group probably have the best curative effect.