UPLC-QE-Orbitrap-Based Cell Metabolomics and Network Pharmacology to Reveal the Mechanism of N-Benzylhexadecanamide Isolated from Maca (Lepidium meyenii Walp.) against Testicular Dysfunction

Abstract

Testicular dysfunction (TDF) is characterized by testosterone deficiency and is caused by oxidative stress injury in Leydig cells. A natural fatty amide named N-benzylhexadecanamide (NBH), derived from cruciferous maca, has been shown to promote testosterone production. Our study aims to reveal the anti-TDF effect of NBH and explore its potential mechanism in vitro. This study examined the effects of H2O2 on cell viability and testosterone levels in mouse Leydig cells (TM3) under oxidative stress. In addition, cell metabolomics analysis based on UPLC-Q-Exactive-MS/MS showed that NBH was mainly involved in arginine biosynthesis, aminoacyl-tRNA biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis, the TCA cycle and other metabolic pathways by affecting 23 differential metabolites, including arginine and phenylalanine. Furthermore, we also performed network pharmacological analysis to observe the key protein targets in NBH treatment. The results showed that its role was to up-regulate ALOX5, down-regulate CYP1A2, and play a role in promoting testicular activity by participating in the steroid hormone biosynthesis pathway. In summary, our study not only provides new insights into the biochemical mechanisms of natural compounds in the treatment of TDF, but also provides a research strategy that integrates cell metabolomics and network pharmacology in order to promote the screening of new drugs for the treatment of TDF.

Exploring the chemical and pharmacological variability of Lepidium meyenii: a comprehensive review of the effects of maca

“…The unstable sulfated compound leads to isothiocyanate formation by the Sulfotransferase (SOT) and Glucosyltransferasa (UGT74) (Blažević et al, 2020). (B) The enzymolysis of the benzylglucosinolate the generation of benzylamine is accomplished by ( Zhang et al, 2023 ). (C) In this example Linoleic acid serves as the scaffold for the formation of 9-hydroperoxy-10E,12Z-octadecadienoic acid and 13-hydroperoxy-9Z,12E-octadecadienoic acid through lipoxygenase enzymes, and then transfers the fundamental skeleton of macaenes using 9 or 13-hydroperoxy-10E,12Z-octadecadienoic acid.…”

“…Network Pharmacology and in silico docking revealed that part of the mechanism of N-Benzylhexadecanamide (NBH). This molecule directly binds and inhibit the CYP1A2 (Cytochrome P450 Family 1 Subfamily A Member 2) active center, which in turn reduces the production of 16α-hydroxydehydro epiandrosterone, preventing excessive consumption of the testosterone (T) precursor DHEA and making it available for T synthesis ( Zang et al, 2023 ).…”

Not All Maca Is Created Equal: A Review of Colors, Nutrition, Phytochemicals, and Clinical Uses

“…Newer research on the science of maca phytochemicals continues, with some recent publications looking at the immunomodulatory [ 98 ] and neuroprotective [ 129 ] aspects of polysaccharides from maca, along with testing isolated compounds like a natural fatty amide known as N -benzylhexadecanamide for its ability to enhance testosterone production [ 130 ]. Using proteomics and lipomics methods, the neuroprotective effects imparted by N -benzylhexadecanamide may be attributed to its effects on sphingolipid metabolism and mitochondrial function, suggesting that it could continue to be investigated for neurodegenerative diseases [ 131 ].…”

Mechanisms of Zhixiao Tang on Anti-Inflammatory Multiple Targets and Multiple Components: Metabonomics Combined with Database Mining Technology

“…6 Consequently, this approach unveils the metabolic pathways and processes of the relevant cells, rendering it highly valuable for in vitro investigations concerning alterations in metabolic profiles and the underlying mechanisms induced by pharmaceutical interventions. 7 In comparison to comprehensive metabolic investigations, cellular metabolomics offers distinct advantages, including the ability to easily regulate experimental variables, achieve high reproducibility, incur low costs, and facilitate result interpretation. Zebrafish (Danio rerio) possesses exceptional attributes such as its diminutive size, prolific reproduction, economical upkeep, brief experimental duration, and substantial homology with mammals, particularly humans.…”