Ascorbic acid as serine protease inhibitor in lung cancer cell line and human serum albumin

Sep 21, 2026

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Serine proteases are widely distributed and their dysregulation during inflammation and infection contributes to conditions including lung and skin inflammation and cancer metastasis. Previous thermodynamic work showed L-ascorbic acid binds trypsin more strongly than pepsin, driven by hydrogen bonds and van der Waals forces, but physiological effects were unclear. This study evaluated ascorbic acid inhibition on serine proteases at multiple concentrations in in-vitro digestion and hydrolysis systems using EDTA-trypsin, SDS-PAGE imaging, and densitometry, revealing dose-dependent inhibition and strong suppression at equal 2.5 mg/mL.

LanguageEnglish

CategoryHealthcare

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Reading Time53 min

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Abstract

Introduction

PLOS ONE OPEN ACCESS Citation: Sil BK, Jamiruddin M. R, Paul PK, Aekwattanaphol N, Nakpheng T, Haq M.A, et al.(2024) Ascorbic acid as serine protease inhibitor in lung cancer cell line and human serum albumin. PLoS ONE 19(7): e0303706 . [https://doi.org/](https://doi.org/)[ ](https://doi.org/)[10.1371/journal.pone](10.1371/journal.pone).0303706 Editor: Faiz ul-Hassan Nasim, Lahore University of Management Sciences, PAKISTAN Received: December 16, 2023 Accepted: April 30, 2024 Published: July 23, 2024 Copyright: © 2024 Sil et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the manuscript and its Supporting Information files. Funding: This research was funded by Reinventing University Grant 2022, Prince of Songkla University, Grant number 175623, awarded to TS. Competing interests: The authors have declared no competing interests. RESEARCH ARTICLE Ascorbic acid as serine protease inhibitor in lung cancer cell line and human serum albumin Bijon Kumar Sil1 *, Mohd. Raeed Jamiruddin2, Pijush Kumar Paul1, Nattanit Aekwattanaphol1, Titpawan Nakpheng1, Md. Ahsanul Haq3, Wilaiporn Buatong1, Teerapol Srichana1 * 1 Drug Delivery System Excellence Center, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla, Thailand, 2 School of Pharmacy, BRAC University, Dhaka, Bangladesh, 3 Immunobiology, Nutrition and Toxicology Lab, Nutrition Research Division, icddr,b, Dhaka, Bangladesh * [teerapol.s@psu.ac.th](teerapol.s@psu.ac.th) (TS); [bkumarsil@gmail.com](bkumarsil@gmail.com) (BKS) Abstract Serine proteases (SPs) are distributed among all living cells accounting for almost one-third of all proteases. Dysregulation of SPs during inflammation and/or infection can result in devastating consequences, such as skin and lung inflammation, neuroinflammation , arthritis, as well as metastasis of cancerous cells. Such activities are tightly regulated by various inhibitors known as serine protease inhibitors (SERPIN) . The thermodynamic investigations previously revealed that L-ascorbic acid binds to trypsin more firmly than pepsin and the binding force of L-ascorbic acid is driven by hydrogen bonds and van der Waals forces. However, the physiochemical effects of such interaction on trypsin and/or pepsin have not yet been reported. Ascorbic acid, also known as vitamin C, is one of the essential nutrients and most common food supplements, fortificants, and preservatives. The aim of this study was to explore the inhibitory effects of ascorbic acid on serine proteases at various concentrations on the in-vitro digestion and/or hydrolysis of intercellular matrix of cell monolayer and human serum albumin (HSA) . The inhibitory effects of ascorbic on trypsin are investigated by qualitative and quantitative analysis using SDS-PAGE imaging and NIH densitometric software. Upon the addition of ascorbic acid in both indicator systems, the detachment and/or dissociation of cell monolayer and the digestion of HSA were inhibited in the presence of EDTA-Trypsin. The inhibitory effect of ascorbic acid on the digestion of intercellular matrix and/or hydrolysis of HSA showed a dose-dependent trend until it reached the maximum extent of inhibition. At an equal concentration (2 .5mg/mL) ascorbic acid and EDTA-Trypsin exhibited the most potent inhibitory effect on the in vitro digestion of protein either in the form of intercellular matrix in cell monolayer and/or HSA respectively. Overall, our results based on two indicator systems strongly indicate that ascorbic acid may function as a serine protease inhibitor (SERPIN) beyond other important functions. 1. Introduction Serine proteases are being targeted for the design of enzyme inhibitors due to their involvement in

FAQs

What was the main aim of the study?

To explore the inhibitory effects of ascorbic acid on serine proteases at various concentrations in in-vitro digestion/hydrolysis systems involving cell monolayers and human serum albumin (HSA).

How were the inhibitory effects on trypsin assessed?

By qualitative and quantitative analysis using SDS-PAGE imaging and NIH densitometric software in the presence of indicator systems including EDTA-trypsin.

Did the inhibition depend on ascorbic acid concentration?

Yes. The inhibitory effect showed a dose-dependent trend until reaching the maximum extent of inhibition, with the most potent effect reported at 2.5 mg/mL alongside EDTA-trypsin.

Sep 21, 2026

Upload by Noah