【摘要】：目的探討丁苯酞的腎保護(hù)機(jī)制及核因子E2相關(guān)因子2(Nrf2)-抗氧化還原反應(yīng)元件(Nrf2-ARE)通路在腎間質(zhì)纖維化中的作用。方法清潔級CD-1小鼠分為4組:假手術(shù)組、模型組、治療組、陽性對照組。假手術(shù)組、模型組均給予等量生理鹽水灌胃。治療組、陽性對照組分別給予丁苯酞、貝那普利灌胃。將各組小鼠分別于術(shù)后第3、7、14天處死,取梗阻側(cè)腎組織,通過免疫組織化學(xué)染色、RT-PCR等實(shí)驗(yàn)手段檢測腎組織Ⅰ型膠原蛋白、Nrf2及γ-GCS蛋白及mRNA的表達(dá)水平。并將Nrf2與γ-GCS及Ⅰ型膠原蛋白與γ-GCS進(jìn)行相關(guān)性分析。結(jié)果免疫組織化學(xué)及RT-PCR結(jié)果均顯示模型組Ⅰ型膠原蛋白mRNA及蛋白較假手術(shù)組有明顯升高,并且隨著梗阻時(shí)間的延長表達(dá)逐漸增多。治療組及陽性對照組小鼠在各時(shí)間點(diǎn)較模型組表達(dá)顯著減少。動(dòng)態(tài)觀察Nrf2mRNA的表達(dá),結(jié)果顯示:與同期假手術(shù)組相比,術(shù)后3、7、14d模型組小鼠腎組織中Nrf2、r-GCS、I型膠原蛋白mRNA表達(dá)均增加,給予丁苯酞干預(yù)后,Nrf2mRNA在7、14d時(shí)治療組與模型組比較表達(dá)均顯著增加(P0.05),治療組Nrf2在各時(shí)間點(diǎn)表達(dá)均強(qiáng)于陽性對照組(P0.05)。RT-PCR學(xué)檢測γ-GCS mRNA變化趨勢同于Nrf2,其腎保護(hù)效應(yīng)隨時(shí)間延長呈增強(qiáng)趨勢。相關(guān)性分析顯示:Nrf2與γ-GCS呈正相關(guān)(r=0.950,P0.05),γ-GCS與Ⅰ型膠原蛋白呈負(fù)相關(guān)(r=-0.629,P0.05)。結(jié)論丁苯酞有抗腎間質(zhì)纖維化作用,機(jī)制可能與激活Nrf2-ARE信號通路,提高腎間質(zhì)中Nrf2、抗氧化酶r-GCS活性水平有關(guān)。
[Abstract]:Objective to investigate the renal protective mechanism of butylphthalide and the role of nuclear factor E2 related factor 2 (Nrf2)-antioxidant reduction element (Nrf2-ARE) pathway in renal interstitial fibrosis. Methods Clean CD-1 mice were divided into 4 groups: false operation group, model group, treatment group and positive control group. In the pseudo-operation group and the model group, the same amount of saline was given intragastrically. In the treatment group and the positive control group, butylphthalide and benazepril were given intragastrically. The mice in each group were killed on the 3rd, 7th and 14th day after operation. The renal tissue of the obstructive side was taken. The expression of type I collagen, Nrf2, gamma-GCS protein and mRNA in renal tissue were detected by immunohistochemical staining and RT-PCR. The correlation between Nrf2 and gamma-GCS and type I collagen and gamma-GCS was analyzed. Results Immunohistochemical and RT-PCR results showed that the expression of type I collagen mRNA and protein in the model group was significantly higher than that in the false operation group, and increased gradually with the prolongation of obstruction time. The expression of mice in treatment group and positive control group was significantly lower than that in model group at each time point. The expression of Nrf2mRNA was observed dynamically. The results showed that the expression of Nrf2,r-GCS, type I collagen mRNA in renal tissue of model group was increased at 3, 7 and 14 days after operation, and the expression of Nrf2mRNA in treatment group was significantly higher than that in model group at 7 and 14 days after butylphthalide intervention (P 0.05). The expression of Nrf2 in the treatment group was stronger than that in the positive control group at all time points (P 0.05). The change trend of 緯-GCS mRNA detected by RT-PCR was the same as the renal protective effect of Nrf2, increased with the prolongation of time. Correlation analysis showed that Nrf2 was positively correlated with 緯-GCS (r 鈮，

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