Screening bioactive compounds from Danggui-shaoyao-san for treating sodium retention in nephrotic syndrome using bio-affinity ultrafiltration.
Screening bioactive compounds from Danggui-shaoyao-san for treating sodium retention in nephrotic syndrome using bio-affinity ultrafiltration.
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DOI：
10.1016/j.jep.2022.115171
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发表时间：
2022-03
影响因子：
5.4
通讯作者：
Mo Yang;Lianghou Ni;Yun-lai Wang;Zihua Xuan;Huan Wu;Wenjing Zhan;Xinyu Wan;Jinghui Wang;Fan Xu
中科院分区：
文献类型：
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作者：
Mo Yang;Lianghou Ni;Yun-lai Wang;Zihua Xuan;Huan Wu;Wenjing Zhan;Xinyu Wan;Jinghui Wang;Fan Xu
民族药理学相关性当归芍药散（DSS）是传统中药活血利水疗法的代表方剂，在临床上已用于治疗肾病综合征（NS）和缓解肾病水肿。拆解为养血-活血(YXHX)和健脾-丽水(JPLS)。用嘌呤霉素氨基核苷（PAN）诱导肾病大鼠模型，并评估DSS、YXHX和JPLS提取物对尿钠排泄、尿纤溶酶（原）含量和纤溶酶活性的影响。体外评价提取物对尿激酶型纤溶酶原激活剂（uPA）和纤溶酶活性的抑制作用。采用生物亲和超滤和高效液相色谱四极杆飞行时间质谱（BAU-UPLC-Q/TOF-MS）快速筛选和定性分析DSS提取物中的uPA/纤溶酶亲和化合物。此外，使用uPA/纤溶酶抑制测定和分子对接来验证潜在生物活性化合物的活性和亲和力机制。结果在体内，DSS、YXHX和JPLS可防止肾病大鼠的钠潴留。 DSS和YXHX处理降低了尿纤溶酶活性，但没有改变尿纤溶酶（原）浓度，并且它们的提取物在体外表现出很强的uPA和纤溶酶抑制活性。这些结果表明，uPA 和纤溶酶是 DSS 和 YXHX 干预 NS 钠潴留的直接靶标。使用 BAU-UPLC-Q/TOF-MS，筛选没食子酸、没食子酸甲酯、albiflorin 和 1,2,3,4,6-O-五没食子酰葡萄糖 (PGG) 作为 uPA 或纤溶酶亲和化合物。其中，PGG被发现是uPA和纤溶酶双重抑制剂，对uPA的IC50为6.861 μM，对纤溶酶的IC50为149.0 μM。 PGG与uPA和纤溶酶的分子对接结果与验证结果一致。结论PAN诱导肾病大鼠的结果表明，通过抑制uPA介导的纤溶酶生成和肾脏纤溶酶活性来干预钠潴留可能是DSS的可能机制。我们得出结论，PGG 是一种潜在的生物活性化合物，与 DSS 对尿钠的影响有关。
Ethnopharmacological relevanceDanggui-shaoyao-san (DSS), a representative formula of Traditional Chinese Medicine (TCM) for promoting blood circulation and diuresis (Huo-Xue-Li-Shui) therapy, has been used to clinically nephrotic syndrome (NS) and relieve nephrotic edema.Aim of the studyTo explore the effects and mechanisms of DSS in improving sodium retention and to identify the bioactive compounds from DSS.Materials and methodsDSS prescriptions were disassembled into Yangxue-Huoxue (YXHX) and Jianpi-Lishui (JPLS). A nephrotic rat model was induced with puromycin aminonucleoside (PAN), and the effects on urinary sodium excretion, urinary plasmin(gen) content, and plasmin activity of DSS, YXHX, and JPLS extracts were assessed. The inhibitory effects on urokinase-type plasminogen activator (uPA) and plasmin activity of extracts were evaluatedin vitro. Bio-affinity ultrafiltration and high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (BAU-UPLC-Q/TOF-MS) were used to rapidly screen and qualitatively analyze the uPA/plasmin affinity compounds from DSS extract. Additionally, uPA/plasmin inhibition assays and molecular docking were used to verify the activity and affinity mechanisms of the potential bioactive compounds.ResultsIn vivo, DSS, YXHX, and JPLS prevented sodium retention in nephrotic rats. DSS and YXHX treatment decreased urinary plasmin activity but did not alter urinary plasmin(ogen) concentration, and their extracts showed strong uPA and plasmin inhibitory activityin vitro. These results suggested that uPA and plasmin are direct targets of DSS and YXHX in intervening NS sodium retention. Using BAU-UPLC-Q/TOF-MS, gallic acids, methyl gallate, albiflorin, and 1,2,3,4,6-O-pentagalloylglucose (PGG) were screened as uPA or plasmin affinity compounds. Among them, PGG was found to be a uPA and plasmin dual inhibitor, with an IC50of 6.861 μM against uPA and an IC50of 149.0 μM against plasmin. The molecular docking results of PGG with uPA and plasmin were consistent with the verification results.ConclusionIntervening in sodium retention by inhibiting uPA-mediated plasmin generation and plasmin activity in the kidneys could be possible mechanisms for DSS, as indicated by the results in PAN-induced nephrotic rats. We conclude that PGG is a potential bioactive compound responsible for the effect of DSS on natriuresis.