Department of Traditional Chinese Medicine, Shandong Provincial Hospital affiliated to Shandong University.
Department of Nephrology, Shandong Provincial Hospital affiliated to Shandong University.
Biosci Trends. 2017;11(6):640-650. doi: 10.5582/bst.2017.01311.
Shenqi detoxification granule (SDG), a traditional Chinese herbal formula, has been shown to have nephroprotective and anti-fibrotic activities in patients with chronic kidney disease (CKD). However, its mechanisms in renal fibrosis and the progression of CKD remain largely unknown. P311, a highly conserved 8-kDa intracellular protein, plays a key role in renal fibrosis by regulating epithelial-mesenchymal transition (EMT). Previously, we found P311 might be involved in the pathogenesis of renal fibrosis by inhibiting EMT via the TGF-β1-Smad-ILK pathway. We also found SDG combined with P311 could ameliorate renal fibrosis by regulating the expression of EMT markers. Here we further examined the effect and mechanism of SDG combined with P311 on TGF-β1-mediated EMT in a rat model of unilateral ureteral occlusion (UUO) renal fibrosis. After establishment of the UUO model successfully, the rats were gavaged with SDG daily and/or injected with recombinant adenovirus p311 (also called Ad-P311) through the tail vein each week for 4 weeks. Serum creatinine (Cr), blood urea nitrogen (BUN) and albumin (ALB) levels were tested to observe renal function, and hematoxylin eosin (HE) and Masson staining were performed to observe kidney histopathology. Furthermore, the expression of EMT markers (E-cadherin and α-smooth muscle actin (α-SMA)) and EMT-related molecules TGF-β1, pSmad2/3, Smad7 and ILK were observed using immunohistochemical staining and Western blot analysis. Treatment with SDG and P311 improved renal function and histopathological abnormalities, as well as reversing the changes of EMT markers and EMT-related molecules, which indicated SDG combined with P311 could attenuate renal fibrosis in UUO rats, and the underlying mechanism might involve TGF-β1-mediated EMT and the TGF-β1-Smad-ILK signaling pathway. Therefore, SDG might be a novel alternative therapy for treating renal fibrosis and delaying the progression of CKD. Furthermore, SDG combined with P311 might have a synergistic effect on attenuating renal fibrosis.
参芪解毒颗粒（SDG）是一种中药配方，已被证明对慢性肾脏病（CKD）患者具有肾保护和抗纤维化作用。然而，其在肾纤维化和 CKD 进展中的机制在很大程度上仍然未知。P311 是一种高度保守的 8kDa 细胞内蛋白，通过调节上皮-间充质转化（EMT）在肾纤维化中发挥关键作用。先前，我们发现 P311 可能通过 TGF-β1-Smad-ILK 途径抑制 EMT 参与肾纤维化的发病机制。我们还发现，SDG 联合 P311 通过调节 EMT 标志物的表达可以改善肾纤维化。在这里，我们进一步研究了 SDG 联合 P311 对单侧输尿管梗阻（UUO）肾纤维化大鼠模型中 TGF-β1 介导的 EMT 的作用和机制。成功建立 UUO 模型后，每天通过灌胃给予大鼠 SDG，并每周通过尾静脉注射重组腺病毒 p311（也称为 Ad-P311）一次，共 4 周。检测血清肌酐（Cr）、血尿素氮（BUN）和白蛋白（ALB）水平以观察肾功能，并进行苏木精-伊红（HE）和 Masson 染色以观察肾脏组织病理学变化。此外，通过免疫组织化学染色和 Western blot 分析观察 EMT 标志物（E-钙黏蛋白和α-平滑肌肌动蛋白（α-SMA））和 EMT 相关分子 TGF-β1、pSmad2/3、Smad7 和 ILK 的表达。SDG 和 P311 的治疗改善了肾功能和组织病理学异常，以及逆转 EMT 标志物和 EMT 相关分子的变化，表明 SDG 联合 P311 可减轻 UUO 大鼠的肾纤维化，其潜在机制可能涉及 TGF-β1 介导的 EMT 和 TGF-β1-Smad-ILK 信号通路。因此，SDG 可能是治疗肾纤维化和延缓 CKD 进展的一种新的替代疗法。此外，SDG 联合 P311 可能对减轻肾纤维化具有协同作用。
