School of Traditional Chinese Medicine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, Liaoning, 110016, PR China.
School of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, Liaoning, 110016, PR China.
J Ethnopharmacol. 2022 Mar 1;285:114870. doi: 10.1016/j.jep.2021.114870. Epub 2021 Nov 27.
Walnut kernel, a well-known TCM, is often used after being defatted in tradition. And defatted walnut powder extract (DWPE) has the actions of tonifying the liver and kidney, dissipating stagnation and removing blood stasis, which has the effect on non-alcoholic fatty liver disease (NAFLD). However, the effective components of DWPE in vivo were unclear and the multiple mechanisms of DWPE against NAFLD have not been explored.
The studies were performed to screen the effective substances in vivo by identification of the metabolites of DWPE in rats and to seek the potential mechanisms of DWPE on NAFLD by construction of the network pharmacology based on metabolites and verification of the highly correlated pathway.
To explore the effective substances in vivo, the metabolites of DWPE were identified in SD rats' bio-samples through UPLC-Q-Exactive Orbitrap MS. To analyze the mechanisms of DWPE on NAFLD, a Metabolite-Target-Disease network was established and the potential mechanisms were predicted. Then, highly correlated pathway was verified in animal and cells studies.
A total of 52 metabolites of DWPE were identified in vivo, which were derived from gallic acid, ellagic acid (EA) and glansreginin A (Gla A). The possible metabolic pathways were phase Ⅰ (hydroxylation, hydrolyzation, etc) and phase Ⅱ metabolic reactions (methylation, sulfation and glucuronidation). Furthermore, in the network pharmacology, 54 core targets were enriched into pathways in cancer, nitrogen metabolism and other 9 pathways, which were essential pathways of DWPE against NAFLD. And the mechanism of nitrogen metabolism was verified in both of animal and cells studies. The results showed that DWPE could decline the concentration of ammonia and increase the expressions of carbonic anhydrase 2 (CA2) and carbamoylphosphate synthetase (CPS1) in nitrogen metabolism.
Taken together, the study revealed the absorption components and their metabolic pathways and demonstrated the mechanism of nitrogen metabolism of DWPE on anti-NAFLD.
桃仁是一种著名的中药，在传统上常经过脱脂后使用。脱脂桃仁粉提取物（DWPE）具有补肝益肾、活血化瘀的作用，对非酒精性脂肪肝（NAFLD）有一定疗效。然而，DWPE 在体内的有效成分尚不清楚，DWPE 治疗 NAFLD 的多种机制尚未得到探索。
本研究通过鉴定 DWPE 在大鼠体内的代谢物，筛选体内有效物质，并基于代谢物构建网络药理学，寻找 DWPE 治疗 NAFLD 的潜在机制，并验证高度相关的通路。
为了探讨体内有效物质，采用 UPLC-Q-Exactive Orbitrap MS 鉴定 DWPE 在 SD 大鼠生物样本中的代谢物。为了分析 DWPE 对 NAFLD 的作用机制，建立代谢物-靶标-疾病网络，预测潜在机制，然后在动物和细胞研究中验证高度相关的通路。
共鉴定出 DWPE 在体内的 52 种代谢物，来源于没食子酸、鞣花酸（EA）和格兰瑞丁 A（Gla A）。可能的代谢途径为Ⅰ相（羟化、水解等）和Ⅱ相代谢反应（甲基化、硫酸化和葡萄糖醛酸化）。此外，在网络药理学中，54 个核心靶标被富集到癌症、氮代谢等 9 条途径中，这些途径是 DWPE 治疗 NAFLD 的关键途径。并且在动物和细胞研究中验证了氮代谢途径。结果表明，DWPE 可降低氨浓度，增加氮代谢中碳酸酐酶 2（CA2）和氨基甲酰磷酸合成酶 1（CPS1）的表达。
综上所述，本研究揭示了 DWPE 的吸收成分及其代谢途径，并证实了 DWPE 抗 NAFLD 作用的氮代谢机制。
