Comparison of the effects and mechanisms of Alismatis Rhizoma with and without salt processing in ameliorating edema due to kidney Yin deficiency based on UPLC-Q-TOF-MS and transcriptomics

Lin Yan, Zemin Ou, Yi Cheng, Yan Tong, Javed Iqbal, Jinyu Wang, Dewen Liu
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Abstract

Abstract Salt-processed Alismatis Rhizoma (SAR) is extensively used in clinical practice and exhibits a more robust urination-promoting effect than Alismatis Rhizoma (AR). This study employed ultra-performance liquid chromatography–quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and pattern analysis to compare the terpenoids between AR and SAR. Additionally, this study compared the effects of AR and SAR on the gene expression in the kidneys of the rat model of edema (syndrome of kidney Yin deficiency) by transcriptomics to decipher the mechanism of salt processing. Materials and methods AR and SAR were extracted by ultrasonication, and data were collected by UPLC-Q-TOF-MS in the positive ion mode. Transcriptome sequencing was employed to determine the gene expression levels of the rat model treated with AR and SAR, and the differentially expressed genes (DEGs) were obtained. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment was performed for the DEGs. The protein-protein interaction (PPI) network was built, on the basis of which the core targets were screened out. Finally, real-time quantitative polymerase chain reaction (RT-PCR) was conducted to verify the core targets. Results A total of 63 terpenoids were detected in AR and SAR, and salt processing had a significant effect on the content of terpenoids. AR and SAR mainly participated in the regulation of inflammation and immune responses, and SAR regulated more DEGs than AR. Additionally, SAR exerted more extensive regulatory effects on the targets than AR. Conclusion Salt processing mainly changes the content of chemical compounds in AR, which may indirectly optimize the proportion of the main compounds to enhance the therapeutic effect while reducing the toxicity. AR and SAR mainly ameliorate the edema due to kidney Yin deficiency by reducing inflammation and improving immunity. Finally, SAR regulates more genes and signaling pathways and exerts more extensive regulatory effects than AR.
基于UPLC-Q-TOF-MS和转录组学比较加盐和不加盐处理的黄精改善肾阴虚水肿的作用和机制
摘要 盐加工的黄连(SAR)被广泛应用于临床,与黄连(AR)相比,SAR具有更强的促进排尿作用。本研究采用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF-MS)和模式分析来比较 AR 和 SAR 的萜类化合物。此外,本研究还通过转录组学比较了 AR 和 SAR 对水肿模型(肾阴虚综合征)大鼠肾脏基因表达的影响,以破译盐处理机制。材料和方法 用超声提取 AR 和 SAR,并用 UPLC-Q-TOF-MS 在正离子模式下收集数据。采用转录组测序法测定 AR 和 SAR 大鼠模型的基因表达水平,并获得差异表达基因(DEGs)。对 DEGs 进行了基因本体(GO)和京都基因组百科全书(KEGG)富集。在此基础上筛选出核心靶标。最后,对核心靶标进行了实时定量聚合酶链反应(RT-PCR)验证。结果 在 AR 和 SAR 中共检测到 63 种萜类化合物,盐处理对萜类化合物的含量有显著影响。AR和SAR主要参与炎症和免疫反应的调控,SAR比AR调控更多的DEGs。此外,SAR 对靶标的调控作用比 AR 更广泛。结论 盐加工主要改变了 AR 中化学成分的含量,从而间接优化了主要化合物的比例,在增强疗效的同时降低了毒性。AR 和 SAR 主要通过消炎和提高免疫力来改善肾阴虚引起的水肿。最后,与 AR 相比,SAR 可调控更多的基因和信号通路,发挥更广泛的调控作用。
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