山奈酚延缓自发性肥胖2型糖尿病小鼠肾损伤研究

The Study of Protective Effects of Kaempferol on Renal Tissues in db/db Mice

目的观察山奈酚(KPF)干预自发肥胖2型糖尿病小鼠(db/db鼠)早期糖尿病肾病,并探讨相关机制。方法 24只健康雄性10周龄db/db鼠随机分为四组:不同剂量KPF给药组(50、200mg/kg)、db/db对照组和二甲双胍(0.2g/kg)阳性对照组,并设db/m对照组。连续灌胃给药10周,检测小鼠血清肌酐(SCr)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α),24小时尿白蛋白排泄率(UAER);留取肾组织,作病理形态学检查、实时定量PCR检测肾组织TNF-α、IL-6的m RNA表达、Western blot检测磷酸化核因子κB p65(p-NF-κBp65)、磷酸化核因子κB抑制蛋白α(p-IκBα)、核因子κB抑制蛋白α激酶(IKKα)蛋白的表达水平。结果不同剂量KPF干预10周,都能使db/db小鼠24小时UAER、SCr、IL-6、TNF-α水平下降,肾组织IL-6、TNF-α基因表达水平明显降低(P<0.01、0.05),p-NF-κBp65、IKKα和p-IκBα蛋白表达水平下降(P<0.01)。结论 KPF可通过抑制核因子κB(NF-κB)介导的炎症反应,对自发2型糖尿病小鼠早期的肾脏损害起到保护作用。

Objective To explore the effects of kaempferol（ KPF） on renal tissues and to elucidate its underlying inflammatory molecular mechanism in db/db mice. Methods Twenty- four male db/db mice were randomly divided into four groups： Two groups were orally administered a low or high dose of KPF（ 50 or 200 mg/kg,respectively）,one control group and one positive control（ metformin hydrochloride 0. 2g/kg） group. Six male db/m mice were set as normal control group. After 10 weeks treatment,24 hour urinary albumin excretion（ UAER） 、SCr were detected; the gene expression of tumor necrosis factor- α（ TNF-α）,and interleukin- 6（ IL- 6） were measured by real time PCR; the protein expression of IKKα、phospho- IκBα、phospho-NF- κBp65 were evaluated by Western blotting. Results After 10 weeks of KPF treatment,the urinary albumin excretion,SCr,IL- 6 and TNF- αwere reduced,the gene expressions of TNF- α and IL- 6 were inhibited. Importantly,the protein expression level of IKKα、phospho- IκBα、phospho- NF- κBp65 in renal tissue was significantly decreased（ P〈0. 01、0. 05）. Conclusions Kaempferol has protective effects on renal tissues through inhibiting NF- κB pathway and related inflammation in db/db mice.