【摘要】 目的探讨丁苯酞的肾保护机制及核因子E2相关因子2(Nrf2)-抗氧化还原反应元件(Nrf2-ARE)通路在肾间质纤维化中的作用。方法清洁级CD-1小鼠分为4组:假手术组、模型组、治疗组、阳性对照组。假手术组、模型组均给予等量生理盐水灌胃。治疗组、阳性对照组分别给予丁苯酞、贝那普利灌胃。将各组小鼠分别于术后第3、7、14天处死,取梗阻侧肾组织,通过免疫组织化学染色、RT-PCR等实验手段检测肾组织Ⅰ型胶原蛋白、Nrf2及γ-GCS蛋白及mRNA的表达水平。并将Nrf2与γ-GCS及Ⅰ型胶原蛋白与γ-GCS进行相关性分析。结果免疫组织化学及RT-PCR结果均显示模型组Ⅰ型胶原蛋白mRNA及蛋白较假手术组有明显升高,并且随着梗阻时间的延长表达逐渐增多。治疗组及阳性对照组小鼠在各时间点较模型组表达显著减少。动态观察Nrf2mRNA的表达,结果显示:与同期假手术组相比,术后3、7、14d模型组小鼠肾组织中Nrf2、r-GCS、I型胶原蛋白mRNA表达均增加,给予丁苯酞干预后,Nrf2mRNA在7、14d时治疗组与模型组比较表达均显著增加(P<0.05),治疗组Nrf2在各时间点表达均强于阳性对照组(P<0.05)。RT-PCR学检测γ-GCS mRNA变化趋势同于Nrf2,其肾保护效应随时间延长呈增强趋势。相关性分析显示:Nrf2与γ-GCS呈正相关(r=0.950,P<0.05),γ-GCS与Ⅰ型胶原蛋白呈负相关(r=-0.629,P<0.05)。结论丁苯酞有抗肾间质纤维化作用,机制可能与激活Nrf2-ARE信号通路,提高肾间质中Nrf2、抗氧化酶r-GCS活性水平有关。更多还原

【Abstract】 Objective To explore the rule of the Nrf2-ARE in renal interstitial fibrosis and the mechanism of butylphthalide on renal protective effect.Methods Seventy-two male CD-1mice were divided into 4groups,Sham group,UUO group,NBP group,ACEI group.The Sham group and UUO group were gavaged with physiological saline.The NBP group was gavaged with butylphthalide.The ACEI group was gavaged with benazepril.After 3,7,14 days,6mice were executed and collection of kidney tissue.The expression of Nrf2,γ-GCS and type Ⅰ collagen were detected by immunohistochemisty,RT-PCR and Western blot.The correlation of Nrf2andγ-GCS was assessmented by linear regression.Results The expression of typeⅠ collagen in UUO group was increased compared with the Sham group,However,the expression of TypeⅠ collagen in NBP group or ACEI group were reduced compared with the UUO group.Compared with Sham group,the expression of Nrf2 mRNA,γ-GCSmRNA and typeⅠ collagen mRNA in uuogroup were increased in 3,7,14 days after surgery.Compared with UUO group at 7th,14 uday,the lelve of Nrf2 mRNA were apparently increased in NBP group(P<0.05).It was a positive correlation between the Nrf2andγ-GCS and negative correlation between theγ-GCS and TypeⅠcollagen.Conclusion The renal protective effect of butylphthalide in the renal interstitial fibrosis was more predominant than benazepril.The roles maybe occurred through increased the expression of Nrf2andγ-GCS and alleviated the expression of TypeⅠ collagen in nephridial tissue.更多还原