Protective effect of soluble protein hydrolysate against H<sub>2</sub>O<sub>2</sub>‑induced intestinal injury: An interventional study

Wei,Jingjing; Tao,Guozhong; Liu,Junlin; Framroze,Bomi; Sylvester,Karl G.

doi:10.3892/mmr.2025.13450

Protective effect of soluble protein hydrolysate against H₂O₂‑induced intestinal injury: An interventional study

Authors:
- Jingjing Wei
- Guozhong Tao
- Junlin Liu
- Bomi Framroze
- Karl G. Sylvester
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Affiliations: Department of Surgery, Stanford University School of Medicine, Stanford, CA 94304, USA, Department of R&D, Hofseth BioCare ASA, Aalesund 6003, Norway
Published online on: January 29, 2025 https://doi.org/10.3892/mmr.2025.13450
Article Number: 85
Copyright: © Wei et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to investigate whether soluble protein hydrolysate (SPH) protects against intestinal oxidative stress injury. An in vitro lactate dehydrogenase assay was used to assess the cytotoxicity and protective effect of SPH. For in vivo assessment, friend virus B NIH Jackson mouse pups aged 21 days were administered with 5% w/v soluble protein hydrolysate (SPH) through drinking water for 14 days and then luminally injected with 0.3% or 0.6% H₂O₂. Thereafter, the fecal samples of mice were collected, and the mice were sacrificed. Intestinal epithelial injury was assessed, and the expressions of 84 oxidative stress‑related genes in intestinal tissues was determined. SPH prophylactically protected against H₂O₂‑induced oxidative stress injury in human intestinal epithelial cells. An animal model of oxidative stress‑induced intestinal injury was established using 0.3 and 0.6% H₂O₂. SPH treatment reduced oxidative stress (0.3% H₂O₂)‑induced gut injury in mice. As no accelerated body growth was observed in SPH‑treated mice, it was hypothesized that the underlying protective mechanism of SPH is not related to nutrient oversupply. Treatment with SPH upregulated five oxidative protective genes that were not consistent between the sexes. Some antioxidative genes, including ferritin heavy polypeptide‑1 (Fth1), heme oxygenase‑1 (Hmox1), NAD(P)H dehydrogenase quinone 1 (Nqo1) and superoxide dismutase 1 (Sod1), were commonly upregulated in both male and female mice. Overall, an antioxidative protective effect was observed following SPH treatment, which may be attributed to the upregulation of genes that protect against oxidative damage. The findings of the present study highlight the promising potential of SPH as a functional food for alleviating intestinal oxidative stress injury.

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1	Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F, Arcoraci V, Squadrito F, Altavilla D and Bitto A: Oxidative stress: Harms and benefits for human health. Oxid Med Cell Longev. 2017:84167632017. View Article : Google Scholar : PubMed/NCBI
2	Bhattacharyya A, Chattopadhyay R, Mitra S and Crowe SE: Oxidative stress: An essential factor in the pathogenesis of gastrointestinal mucosal diseases. Physiol Rev. 94:329–354. 2014. View Article : Google Scholar : PubMed/NCBI
3	Rajendran P, Nandakumar N, Rengarajan T, Palaniswami R, Gnanadhas EN, Lakshminarasaiah U, Gopas J and Nishigaki I: Antioxidants and human diseases. Clin Chim Acta. 436:332–347. 2014. View Article : Google Scholar : PubMed/NCBI
4	Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M and Telser J: Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol. 39:44–84. 2007. View Article : Google Scholar : PubMed/NCBI
5	Loboda A, Damulewicz M, Pyza E, Jozkowicz A and Dulak J: Role of Nrf₂/HO-1 system in development, oxidative stress response and diseases: An evolutionarily conserved mechanism. Cell Mol Life Sci. 73:3221–3247. 2016. View Article : Google Scholar : PubMed/NCBI
6	Wu Q, Zhang XS, Wang HD, Zhang X, Yu Q, Li W, Zhou ML and Wang XL: Astaxanthin activates nuclear factor erythroid-related factor 2 and the antioxidant responsive element (Nrf2-ARE) pathway in the brain after subarachnoid hemorrhage in rats and attenuates early brain injury. Mar Drugs. 12:6125–6141. 2014. View Article : Google Scholar : PubMed/NCBI
7	Talalay P, Dinkova-Kostova AT and Holtzclaw WD: Importance of phase 2 gene regulation in protection against electrophile and reactive oxygen toxicity and carcinogenesis. Adv Enzyme Regul. 43:121–134. 2003. View Article : Google Scholar : PubMed/NCBI
8	Mirończuk-Chodakowska I, Witkowska AM and Zujko ME: Endogenous non-enzymatic antioxidants in the human body. Adv Med Sci. 63:68–78. 2018. View Article : Google Scholar : PubMed/NCBI
9	Kiewiet MBG, Faas MM and de Vos P: Immunomodulatory protein hydrolysates and their application. Nutrients. 10:9042018. View Article : Google Scholar : PubMed/NCBI
10	Parolini C, Vik R, Busnelli M, Bjørndal B, Holm S, Brattelid T, Manzini S, Ganzetti GS, Dellera F, Halvorsen B, et al: A salmon protein hydrolysate exerts lipid-independent anti-atherosclerotic activity in ApoE-deficient mice. PLoS One. 9:e975982014. View Article : Google Scholar : PubMed/NCBI
11	Espe M, Holen E, He J, Provan F, Chen L, Øysæd KB and Seliussen J: Hydrolyzed fish proteins reduced activation of caspase-3 in H2O2 induced oxidative stressed liver cells isolated from atlantic salmon (salmo salar). Springerplus. 4:6582015. View Article : Google Scholar : PubMed/NCBI
12	Idowu AT, Benjakul S, Sinthusamran S, Sookchoo P and Kishimura H: Protein hydrolysate from salmon frames: Production, characteristics and antioxidative activity. J Food Biochem. 43:e127342019. View Article : Google Scholar : PubMed/NCBI
13	Wei J, Tao G, Xu B, Wang K, Liu J, Chen CH, Dunn JCY, Currie C, Framroze B and Sylvester KG: Soluble protein hydrolysate ameliorates gastrointestinal inflammation and injury in 2,4,6-trinitrobenzene sulfonic acid-induced colitis in mice. Biomolecules. 12:12872022. View Article : Google Scholar : PubMed/NCBI
14	National Research Council (US) Committee for the Update of the Guide for the Care and Use of Laboratory Animals, . Guide for the Care and Use of Laboratory Animals. 8th Edition. National Academies Press; Washington, DC: 2011
15	McElroy SJ, Castle SL, Bernard JK, Almohazey D, Hunter CJ, Bell BA, Al Alam D, Wang L, Ford HR and Frey MR: The ErbB4 ligand neuregulin-4 protects against experimental necrotizing enterocolitis. Am J Pathol. 184:2768–2778. 2014. View Article : Google Scholar : PubMed/NCBI
16	Wang K, Tao G, Sun Z, Wei J, Liu J, Taylor J, Gibson M, Mostaghimi M, Good M and Sylvester KG: Fecal keratin 8 is a noninvasive and specific marker for intestinal injury in necrotizing enterocolitis. J Immunol Res. 2023:53566462023. View Article : Google Scholar : PubMed/NCBI
17	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−delta delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
18	Imam MU, Zhang S, Ma J, Wang H and Wang F: Antioxidants mediate both iron homeostasis and oxidative stress. Nutrients. 9:6712017. View Article : Google Scholar : PubMed/NCBI
19	Orino K, Lehman L, Tsuji Y, Ayaki H, Torti SV and Torti FM: Ferritin and the response to oxidative stress. Biochem J. 357:241–247. 2001. View Article : Google Scholar : PubMed/NCBI
20	Wang CY and Chau LY: Heme oxygenase-1 in cardiovascular diseases: Molecular mechanisms and clinical perspectives. Chang Gung Med J. 33:13–24. 2010.PubMed/NCBI
21	Correa-Costa M, Amano MT and Câmara NO: Cytoprotection behind heme oxygenase-1 in renal diseases. World J Nephrol. 1:4–11. 2012. View Article : Google Scholar : PubMed/NCBI
22	Naito Y, Takagi T, Uchiyama K and Yoshikawa T: Heme oxygenase-1: A novel therapeutic target for gastrointestinal diseases. J Clin Biochem Nutr. 48:126–133. 2011. View Article : Google Scholar : PubMed/NCBI
23	Chang M, Xue J, Sharma V and Habtezion A: Protective role of hemeoxygenase-1 in gastrointestinal diseases. Cell Mol Life Sci. 72:1161–1173. 2015. View Article : Google Scholar : PubMed/NCBI
24	Schulz S, Wong RJ, Jang KY, Kalish F, Chisholm KM, Zhao H, Vreman HJ, Sylvester KG and Stevenson DK: Heme oxygenase-1 deficiency promotes the development of necrotizing enterocolitis-like intestinal injury in a newborn mouse model. Am J Physiol Gastrointest Liver Physiol. 304:G991–G1001. 2013. View Article : Google Scholar : PubMed/NCBI
25	Schulz S, Chisholm KM, Zhao H, Kalish F, Yang Y, Wong RJ and Stevenson DK: Heme oxygenase-1 confers protection and alters T-cell populations in a mouse model of neonatal intestinal inflammation. Pediatr Res. 77:640–648. 2015. View Article : Google Scholar : PubMed/NCBI
26	Eisenstein RS, Garcia-Mayol D, Pettingell W and Munro HN: Regulation of ferritin and heme oxygenase synthesis in rat fibroblasts by different forms of iron. Proc Natl Acad Sci USA. 88:688–692. 1991. View Article : Google Scholar : PubMed/NCBI
27	Cheng HT, Yen CJ, Chang CC, Huang KT, Chen KH, Zhang RY, Lee PY, Miaw SC, Huang JW, Chiang CK, et al: Ferritin heavy chain mediates the protective effect of heme oxygenase-1 against oxidative stress. Biochim Biophys Acta. 1850:2506–2517. 2015. View Article : Google Scholar : PubMed/NCBI
28	Chen H, Tang X, Zhou B, Zhou Z, Xu N and Wang Y: A ROS-mediated mitochondrial pathway and Nrf₂ pathway activation are involved in BDE-47 induced apoptosis in Neuro-2a cells. Chemosphere. 184:679–686. 2017. View Article : Google Scholar : PubMed/NCBI
29	Hu YR, Ma H, Zou ZY, He K, Xiao YB, Wang Y, Feng M, Ye XL and Li XG: Activation of Akt and JNK/Nrf₂/NQO1 pathway contributes to the protective effect of coptisine against AAPH-induced oxidative stress. Biomed Pharmacother. 85:313–322. 2017. View Article : Google Scholar : PubMed/NCBI
30	Hu XM, Wang YM, Zhao YQ, Chi CF and Wang B: Antioxidant peptides from the protein hydrolysate of monkfish (LOPHIUS LITULON) muscle: Purification, identification, and cytoprotective function on HepG₂ cells damage by H₂O₂. Mar Drugs. 18:1532020. View Article : Google Scholar : PubMed/NCBI
31	Zhao Y, Li H, Men LL, Huang RC, Zhou HC, Xing Q, Yao JJ, Shi CH and Du JL: Effects of selenoprotein S on oxidative injury in human endothelial cells. J Transl Med. 11:2872013. View Article : Google Scholar : PubMed/NCBI
32	Ye Y, Fu F, Li X, Yang J and Liu H: Selenoprotein S is highly expressed in the blood vessels and prevents vascular smooth muscle cells from apoptosis. J Cell Biochem. 117:106–117. 2016. View Article : Google Scholar : PubMed/NCBI
33	Li X, Chen M, Yang Z, Wang W, Lin H and Xu S: Selenoprotein S silencing triggers mouse hepatoma cells apoptosis and necrosis involving in intracellular calcium imbalance and ROS-mPTP-ATP. Biochim Biophys Acta Gen Subj. 1862:2113–2123. 2018. View Article : Google Scholar : PubMed/NCBI
34	Okina Y, Sato-Matsubara M, Matsubara T, Daikoku A, Longato L, Rombouts K, Thanh Thuy LT, Ichikawa H, Minamiyama Y, Kadota M, et al: TGF-β1-driven reduction of cytoglobin leads to oxidative DNA damage in stellate cells during non-alcoholic steatohepatitis. J Hepatol. 73:882–895. 2020. View Article : Google Scholar : PubMed/NCBI
35	Lv Y, Wang Q, Diao Y and Xu R: Cytoglobin: A novel potential gene medicine for fibrosis and cancer therapy. Curr Gene Ther. 8:287–294. 2008. View Article : Google Scholar : PubMed/NCBI
36	Yu C and Xiao JH: The Keap1-Nrf2 system: A mediator between oxidative stress and aging. Oxid Med Cell Longev. 2021:66354602021. View Article : Google Scholar : PubMed/NCBI
37	Chevrier G, Mitchell PL, Rioux LE, Hasan F, Jin T, Roblet CR, Doyen A, Pilon G, St-Pierre P, Lavigne C, et al: Low-molecular-weight peptides from salmon protein prevent obesity-linked glucose intolerance, inflammation, and dyslipidemia in LDLR-/-/ApoB100/100 mice. J Nutr. 145:1415–1422. 2015. View Article : Google Scholar : PubMed/NCBI