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目的:动态观察黄芪(AM)对单侧输尿管梗阻(UUO)所致大鼠肾小管问质纤维化的拮抗作用及对转化生长因子β1(TGF-β1)表达的影响,初步探讨其可能的分子作用机制。方法:SD大鼠随机分为假手术组、UUO模型组和黄芪治疗组。在造模后3,7,14 d处死动物。观察肾脏病理改变。采用免疫组化方法检测肾组织中TGF-β1和α-平滑肌肌动蛋白(α-SMA)表达。用实时荧光定量RT-PCR检测大鼠TGF-β1 mRNA和α-SMA mRNA表达量。用Western blot检测大鼠TGF-β1蛋白表达量。结果:与假手术组相比,各时间点的UUO大鼠肾脏病理损害进行性加重,TGF-β1和α-SMA mRNA和蛋白表达随梗阻时间延长逐渐增高;黄芪治疗后可明显减轻UUO大鼠肾组织的肾小管损伤及肾间质纤维化,使UUO大鼠肾组织中高表达的TGF-β1和α-SMA降低(P<0.05)。结论:黄芪可能通过从核酸和蛋白水平抑制TGF-β1的表达,抑制肾小管上皮细胞转分化,减轻和改善UUO大鼠肾间质纤维化。
OBJECTIVE: To dynamically observe the antagonism of Astragalus membranaceus (AM) on tubulointerstitial fibrosis induced by unilateral ureteral obstruction (UUO) and the effect of transforming growth factor-β1 (TGF-β1) on the expression of transforming growth factor β1 Mechanism. Methods: SD rats were randomly divided into sham operation group, UUO model group and Astragalus treatment group. Animals were sacrificed 3, 7 and 14 days after modeling. Observe the renal pathological changes. Immunohistochemistry was used to detect the expression of TGF-β1 and α-smooth muscle actin (α-SMA) in renal tissue. Real-time fluorescent quantitative RT-PCR was used to detect the mRNA expression of TGF-β1 and α-SMA in rats. Western blot was used to detect the expression of TGF-β1 in rats. Results: Compared with the sham operation group, the pathological changes of kidney in UUO rats were progressively worsened at each time point. The mRNA and protein expressions of TGF-β1 and α-SMA gradually increased with the prolongation of obstruction time. Astragalus membranaceus treatment could significantly reduce the UUO rats Renal tubular damage and renal interstitial fibrosis, UUO rats kidney tissue high expression of TGF-β1 and α-SMA decreased (P <0.05). Conclusion: Astragalus may inhibit renal tubular epithelial cell transdifferentiation by inhibiting the expression of TGF-β1 from nucleic acid and protein level, and reduce and improve renal interstitial fibrosis in UUO rats.