柴胡皂苷 B2 通过抑制 Hedgehog 通路减轻肾脏纤维化。
Saikosaponin B2 attenuates kidney fibrosis via inhibiting the Hedgehog Pathway.
机构信息
Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, PR China.
Laboratory of Medicinal Plant Biotechnology, College of Pharmacy, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, PR China.
出版信息
Phytomedicine. 2020 Feb;67:153163. doi: 10.1016/j.phymed.2019.153163. Epub 2019 Dec 28.
BACKGROUND
Renal interstitial fibrosis is a common pathway through which chronic kidney disease progresses to end-stage renal disease. There are currently no effective drugs available to treat kidney fibrosis, so traditional medicine is likely to be a candidate. The therapeutic potential of saikosaponin B2 (SSB2), a biologically active ingredient of Radix Bupleuri, on renal fibrosis has not been reported.
METHODS
A unilateral ureteral obstruction (UUO) model was conducted to induce renal interstitial fibrosis in mice. SSB2's effect was valuated by histological staining and exploring the changes in expression of relative proteins and mRNAs. A conditional medium containing sonic hedgehog variant protein stimulating normal rat kidney interstitial fibroblast cells (NRK-49F) was used in an in vitro model to determine the possible mechanism. The molecular target of SSB2 was verified using several mutation plasmids.
RESULTS
SSB2 administration reduced kidney injury and alleviated interstitial fibrosis by decreasing excessive accumulation of extracellular matrix components in UUO mice. It could also reduce the expression of α-SMA, fibronectin and Gli1, a crucial molecule of the hedgehog (Hh) signaling pathway both in vivo and in vitro. In NIH-3T3 cells simulated by conditional medium containing sonic hedgehog variant protein, SSB2 showed the ability to decrease the expression of Gli1 and Ptch1 mRNA. Using a dual-luciferase reporter assay, SSB2 suppressed the Gli-luciferase reporter activity in NIH-3T3 cells, and the IC was 0.49 μM, but had no effect on the TNF-α/NF-κB and Wnt/β-catenin signaling pathways, indicating the inhibition selectivity on the Hh signaling pathway. Furthermore, SSB2 failed to inhibit the Hh pathway activity evoked by ectopic expression of Gli2ΔN and Smo D473H, suggesting that SSB2 might potentially act on smoothened receptors.
CONCLUSION
SSB2 could attenuate renal fibrosis and decrease fibroblast activation by inhibiting the Hh signaling pathway.
背景
肾间质纤维化是慢性肾脏病进展至终末期肾病的常见途径。目前尚无有效的药物可用于治疗肾纤维化，因此传统医学可能是一个候选药物。柴胡皂苷 B2（SSB2）是柴胡的一种生物活性成分，其在肾纤维化方面的治疗潜力尚未有报道。
方法
通过单侧输尿管梗阻（UUO）模型在小鼠中诱导肾间质纤维化。通过组织学染色和探索相关蛋白和 mRNA 表达的变化来评估 SSB2 的作用。在体外模型中，使用含有 sonic hedgehog 变体蛋白刺激正常大鼠肾间充质成纤维细胞（NRK-49F）的条件培养基来确定可能的机制。使用几种突变质粒验证 SSB2 的分子靶标。
结果
SSB2 给药通过减少 UUO 小鼠细胞外基质成分的过度积累，减轻了肾脏损伤并缓解了间质纤维化。它还可以降低 Hedgehog（Hh）信号通路中的关键分子α-SMA、纤维连接蛋白和 Gli1 在体内和体外的表达。在含有 sonic hedgehog 变体蛋白的条件培养基模拟的 NIH-3T3 细胞中，SSB2 表现出降低 Gli1 和 Ptch1 mRNA 表达的能力。使用双荧光素酶报告基因检测，SSB2 抑制了 NIH-3T3 细胞中的 Gli-荧光素酶报告基因活性，IC 为 0.49 μM，但对 TNF-α/NF-κB 和 Wnt/β-catenin 信号通路没有影响，表明对 Hh 信号通路具有抑制选择性。此外，SSB2 未能抑制Gli2ΔN 和 Smo D473H 异位表达引起的 Hh 通路活性，表明 SSB2 可能潜在地作用于 smoothened 受体。
结论
SSB2 可通过抑制 Hh 信号通路来减轻肾纤维化并减少成纤维细胞激活。
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