Heilongjiang University of Traditional Chinese Medicine, Heping Road 24, Harbin, 150040, China.
Heilongjiang University of Traditional Chinese Medicine, Heping Road 24, Harbin, 150040, China; Guangdong Pharmaceutical University, Huandong Road 280, Guangzhou, 510006, China.
J Pharm Biomed Anal. 2020 Sep 5;188:113336. doi: 10.1016/j.jpba.2020.113336. Epub 2020 May 22.
Phellodendri Amurensis cortex (PAC), a famous traditional Chinese herb with good anti-inflammatory efficacy, is used to treat various liver and kidney sickness in clinical practice. However, the potential mechanisms protecting against nephritis of PAC have not been comprehensively elucidated. The aim of this research was to explore the mechanism of PAC against doxorubicin- induced nephritis in rats by characterizing metabolites and potential metabolic pathways changes. The rat models of nephritis were established using 6.5 mg/kg doxorubicin injection from caudal vein for five weeks. The rats in the treatment group were respectively received PAC extract at the dose of 216, 432, and 864 mg/kg once a day during the experiment. Then, urine metabolomics strategy based on ultra-performance liquid chromatography-quadrupole time of flight-tandem mass spectrometry (UPLC-Q/TOF-MS) has been employed to discover the possible significant metabolites and metabolic pathway of nephritis rats. At the end of the experiment, serum, urine and kidney tissue were collected for biochemical and pathological examination. The results showed that PAC treatment notably decreased urinary protein, serum Cr content and renal tissue lesions, and increased serum TP and ALB content. A total of potential twenty- eight metabolites such as 5'-methylthioadenosine, cGMP, dehydroepiandrosterone sulfate, salbuta, 2-phenylaminoadenosine contributing to nephritis rat model were selected and identified in the urine samples. Compared with the model group, the high-dose PAC group can recall 18 metabolites level, the medium-dose group can recall 13 metabolites level, and the low-dose PAC group can recall 8 metabolites level, which were involved in nine primary metabolic pathways such as steroid hormone biosynthesis, alanine,aspartate and glutamate metabolism, cysteine and methionine metabolism as well as glyoxylate and dicarboxylate metabolism. The protein expressions of key enzymes involving methylthioadenosine phosphorylase (Mtap), cytidine deaminase (Cda), thymidine kinase (Tk), argininosuccinate synthase (Ass) in metabolic pathways were further verified by Western blot. The results showed that Phellodendron chinense up-regulated the protein expressions of Cda and Tk and down-regulated the protein expressions of Mtap and Ass. In conclusion, PAC possesses renoprotective effect against doxorubicin-induced nephritis, which may be mediated via regulating differential metabolites, reducing oxidative stress response, improving renal function, enhancing the ability of the immune system, regulating the role of key enzymes.
黄柏（PAC）是一种著名的传统中药，具有良好的抗炎功效，在临床实践中用于治疗各种肝肾疾病。然而，PAC 防治肾炎的潜在机制尚未得到全面阐明。本研究旨在通过研究特征代谢物和潜在代谢途径的变化，探讨 PAC 对阿霉素诱导的大鼠肾炎的作用机制。采用尾静脉注射 6.5mg/kg 阿霉素 5 周建立大鼠肾炎模型。治疗组大鼠分别给予 PAC 提取物 216、432 和 864mg/kg，每天 1 次。然后，采用基于超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q/TOF-MS）的尿液代谢组学策略，发现肾炎大鼠的可能显著代谢物和代谢途径。实验结束时，收集血清、尿液和肾组织进行生化和病理检查。结果表明，PAC 治疗可显著降低尿蛋白、血清 Cr 含量和肾组织病变，增加血清 TP 和 ALB 含量。在尿液样本中共鉴定出潜在的 28 种代谢物，如 5'-甲基硫代腺苷、cGMP、脱氢表雄酮硫酸酯、沙丁胺醇、2-苯氨基腺苷等，这些代谢物与肾炎大鼠模型有关。与模型组相比，高剂量 PAC 组可使 18 种代谢物水平恢复，中剂量 PAC 组可使 13 种代谢物水平恢复，低剂量 PAC 组可使 8 种代谢物水平恢复，这些代谢物涉及类固醇激素生物合成、丙氨酸、天冬氨酸和谷氨酸代谢、半胱氨酸和蛋氨酸代谢以及乙醛酸和二羧酸代谢等 9 个主要代谢途径。Western blot 进一步验证了代谢途径中关键酶的蛋白表达，包括甲基硫代腺苷磷酸化酶（Mtap）、胞嘧啶脱氨酶（Cda）、胸苷激酶（Tk）、精氨酸代琥珀酸合成酶（Ass）。结果表明，黄柏上调了 Cda 和 Tk 的蛋白表达，下调了 Mtap 和 Ass 的蛋白表达。综上所述，黄柏对阿霉素诱导的肾炎具有肾保护作用，其机制可能是通过调节差异代谢物、减轻氧化应激反应、改善肾功能、增强免疫系统能力、调节关键酶的作用来实现的。
