Walnut Kernel Oil and Defatted Extracts Enhance Mesenchymal Stem Cell Stemness and Delay Senescence

Marwa A Elsied; Zeina W Sharawi; Hadba Al-Amrah; Rabab A Hegazy; Amro E Mohamed; Rasha M Saleh; Sanad S El-Kholy; Foad A Farrag; Masoud H Fayed; Mohammed A El-Magd

doi:10.3390/molecules28176281

Walnut Kernel Oil and Defatted Extracts Enhance Mesenchymal Stem Cell Stemness and Delay Senescence

Molecules. 2023 Aug 28;28(17):6281. doi: 10.3390/molecules28176281.

Authors

Affiliations

¹ Anatomy and Embryology Department, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafr El-Sheikh 33516, Egypt.
² Biological Sciences Department, Faculty of Sciences, King AbdulAziz University, Jeddah 21589, Saudi Arabia.
³ Department of Biology, Al-Darb University College, Jazan University, Jazan 45142, Saudi Arabia.
⁴ Biochemistry Division, Chemistry Department, Faculty of Science, Tanta University, Tanta 31527, Egypt.
⁵ Department of Physiology, Faculty of Veterinary Medicine, Mansura University, Mansura 35516, Egypt.
⁶ Department of Physiology, Faculty of Medicine, Kafrelsheikh University, Kafr El-Shaikh 33516, Egypt.

Abstract

Decreased stemness and increased cellular senescence impair the ability of mesenchymal stem cells (MSCs) to renew themselves, change into different cell types, and contribute to regenerative medicine. There is an urgent need to discover new compounds that can boost MSCs' stemness and delay senescence. Therefore, this study aimed to investigate the impact of walnut kernel oil (WKO) and defatted (WKD) extracts on bone marrow (BM)-MSC stemness and senescence. Premature senescence and inflammation were induced in BM-MSCs using H₂O₂ and LPS, respectively. Phytochemical constituents of WKO and WKD extracts were detected by HPLC. The stemness (proliferation and migration), senescence-related markers (p53, p21, SIRT1, and AMPK), oxidative stress/antioxidant markers, inflammatory cytokines, and cell cycle of BM-MSCs were measured by MTT assay, qPCR, ELISA, and flow cytometry. WKO and WKD extracts improved rat BM-MSC stemness, as evidenced by (1) increased cell viability, (2) decreased apoptosis (low levels of Bax and caspase3 and high levels of Bcl2), (3) upregulated MMP9 and downregulated TIMP1 expression, and (4) cell cycle arrest in the G0/G1 phase and declined cell number in the S and G2/M phases. Additionally, WKO and WKD extracts reduced rat BM-MSC senescence, as indicated by (1) decreased p53 and p21 expression, (2) upregulated expression and levels of SIRT1 and AMPK, (3) reduced levels of ROS and improved antioxidant activity (higher activity of CAT, SOD, and GPx and upregulated expression of NrF2 and HO-1), and (4) declined levels of TNFα, IL1β, and NF-κB. When compared to the WKO extract, the WKD extract had a greater impact on the induction of stemness and reduction of senescence of BM-MSCs due to its stronger antioxidant activity, which could be attributed to its higher levels of flavonoids and phenolic compounds, as detected by HPLC analysis. WKO and WKD extracts enhance rat BM-MSC stemness and protect them from senescence, suggesting their potential use as enhancers to increase MSCs' therapeutic efficacy.

Keywords: antioxidant; mesenchymal stem cells; senescence; stemness; walnut kernel extracts.

MeSH terms

AMP-Activated Protein Kinases*
Animals
Antioxidants / pharmacology
Hydrogen Peroxide
Juglans*
Rats
Sirtuin 1 / genetics
Tumor Suppressor Protein p53

Substances

AMP-Activated Protein Kinases
Antioxidants
Hydrogen Peroxide
Sirtuin 1
Tumor Suppressor Protein p53

Grants and funding

The authors did not receive any external funding.