Grape seed extract and L-ascorbic acid exert anti-neoplastic effects against solid Ehrlich carcinoma in vivo through modulating the tumor microenvironment and Th1/Th2 balance

Dalia Sami Morsi¹Heba M. R. Hathout²Hind S. AboShabaan¹Mahmoud Emam³Manal Elkhadragy⁴Ahmed Esmat Abdel Moneim^5*Islam M. El-Garawani¹Hagar A. Abu Quora¹

• ¹Menoufia University, Shebeen ElKom, Egypt
• ²Cairo University, Giza, Egypt
• ³National Research Centre, Ad Doqi, Egypt
• ⁴Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia
• ⁵Helwan University, Helwan, Egypt

The existing study sought to highlight the modulatory effect of co-treatment based on grape seed extract (GSE) and L. ascorbic acid (AA) on tumor microenvironment and immune response in murine solid Ehrlich carcinoma (SEC). Herein, GSE (200 mg/kg; orally) and AA (50 mg/kg; orally) were given either separately or in combination for 14 days. GSE active metabolites were identified using GC-MS and LC-MS/MS. Tumor size, Ki-67, Caspase-3, intratumoral infiltrated CD4 + , CD8 + , and FOXP3 + cells were detected immunohistochemically. Oxidative stress in tumor cells was determined. Serum levels of IL-12, IFN-γ, IL-4, and IL-10 were detected using ELISA. The results revealed that treatment with GSE and/or AA markedly diminished tumor size, intensified intratumoral oxidative stress, downregulated tumor cell proliferation along with upregulated tumor cells' apoptosis. GSE and AA enhanced tumor immune microenvironment through increasing CD8 + and CD4 + T cells accompanied by decreasing FOXP3 + Treg cells infiltrated in tumors. GSE and/or AA moved Th1/Th2 balance in favor of Th1 as evidenced by increased serum levels of IFN-γ and IL-12 accompanied with decreased serum levels of IL-4 and IL-10. These findings may be attributed to the presence of different chemical scaffolds of phenolic acids, Flavan-3-ols and its glycosides, glycerolipids and its glycosides, glycosylated seco-iridoids, dihydrochalcone, stilbenoid, flavone, dihydroxyflavone, and methylated flavone, sugars, and fatty acids. In conclusion, results suggested that dual treatment based on GSE & AA are promising anticancer therapeutics, through their potency to control proliferation, induce apoptosis, intratumoral oxidative stress, modulate tumor immune microenvironment, and shifting Th1/Th2 response toward Th1.