葛根素对急性肾损伤的改善作用及相关机制

EFFECT OF PUERARIN IN IMPROVING ACUTE KIDNEY INJURY AND RELATED MECHANISMS

目的探讨葛根素对脂多糖(LPS)诱导小鼠急性肾损伤的改善作用及其相关机制。方法将24只BALB/C小鼠随机分为4组,葛根素组以50 mg/kg葛根素灌胃,地塞米松组(阳性对照)腹腔注射5 mg/kg的地塞米松,模型组和对照组灌胃给予同等容量的生理盐水。葛根素组和地塞米松组小鼠预防性给药1 h后,腹腔注射10 mg/kg LPS刺激12 h。采用苏木精-伊红(HE)染色观察各组小鼠肾组织形态学改变,同时检测肾功能指标血尿素氮(BUN)和肌酐(Cr)水平;采用酶联免疫吸附试验(ELISA)法检测小鼠血清中炎症因子肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的含量,Western blotting法检测肾组织中炎症蛋白诱导型一氧化氮合酶(iNOS)和环氧合酶2(COX2)的表达。结果与对照组比较,LPS诱导急性肾损伤小鼠肾组织中可见肾小管坏死中心的产生,血清中BUN和Cr的水平显著升高(F=51.55、102.75,P<0.01),血清中炎症因子TNF-α和IL-6的含量显著增加(F=3338.15、1226.85,P<0.01),肾组织中炎症蛋白iNOS和COX2的表达显著升高(F=21682.77、4497.00,P<0.01);葛根素预防给药1 h能显著改善LPS引发的上述变化(P<0.01)。结论葛根素对LPS诱导的急性肾损伤具有明显的改善作用,同时可明显减少炎症因子的释放及降低炎症蛋白的表达,提示抗炎可能是葛根素治疗急性肾损伤的药理学基础。

Objective To investigate the effect of puerarin in improving lipopolysaccharide(LPS)-induced acute kidney injury and related mechanisms.Methods A total of 24 BALB/C mice were randomly divided into puerarin group(50 mg/kg puerarin by gavage),dexamethasone(positive control)group(intraperitoneal injection of 5 mg/kg dexamethasone),model group(an equal volume of normal saline by gavage),and control group(an equal volume of normal saline by gavage).At 1 hour after prophylactic administration,the mice in the puerarin group and the dexamethasone group were given intraperitoneal injection of LPS(10 mg/kg)and were stimulated for 12 hours.Hematoxylin-eosin staining was used to observe the morphological changes of renal tissue,and the levels of the renal function markers blood urea nitrogen(BUN)and creatinine(Cr)were also measured.ELISA was used to measure the serum level of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6),and Western blotting was used to measure the protein expression of inducible nitric oxide synthase(iNOS)and cyclooxygenase-2(COX2)in renal tissue.Results Compared with the control group,the mice with LPS-induced acute kidney injury showed the formation of renal tubular necrosis in renal tissue and significant increases in the serum levels of BUN and Cr(F=51.55,102.75;P<0.01),the serum levels of the inflammatory factors TNF-αand IL-6(F=3338.15,1226.85;P<0.01),and the protein expression levels of iNOS and COX2 in renal tissue(F=21682.77,4497.00;P<0.01).Prophylactic administration of puerarin for 1 hour significantly improved the above changes induced by LPS(P<0.01).Conclusion Puerarin can significantly improve LPS-induced acute kidney injury and reduce the release of inflammatory factors and the expression of inflammatory proteins,suggesting that anti-inflammation is the pharmacological basis of puerarin in the treatment of acute kidney injury.