Evaluation of the Antiglycation Effect of Branched Chain Amino Acids and Phytochemical Compounds on RAW 264.7 Cell Line and their Synergistic Effect on Colorectal Cancer Cell Line Panel



  • Moath Alqaraleh Pharmacological and Diagnostic Research Center (PDRC), Faculty of Pharmacy, Al-Ahliyya Amman University, Amman 19328, Jordan
  • Violet Kasabri Department of Pharmacy, Faculty of Pharmacy. The University of Jordan, Amman, Jordan
  • Ahmad Aljaafreh Department of Medical Analysis, Mutah University, Mutah, Karak, 61710, Jordan
  • Ibrahim Al-Majali Department of Medical Analysis, Mutah University, Mutah, Karak, 61710, Jordan
  • Nihad Al-Othman Division of Anatomy, Biochemistry and, Genetics. Faculty of Medicine and Health Sciences, An-Najah National University, Nablus
  • Khaled Khleifat Department of Biology, Mutah University, Karak, Mutah, 61710 Jordan
  • Sara Khaleel Faculty of Pharmacy, Zarqa University, P.O. Box 132222, Zarqa, 13132, Jordan
  • Nafe M. Al‐Tawarah Department of Medical Analysis, Mutah University, Mutah, Karak, 61710, Jordan
  • Haitham Qaralleh Department of Medical Analysis, Mutah University, Mutah, Karak, 61710, Jordan
  • Mohammad Al-majali Department of Biological Sciences, Faculty of Science. The University of Jordan, Amman, Jordan
  • Israa Yousef Pharmacological and Diagnostic Research Center (PDRC), Faculty of Pharmacy, Al-Ahliyya Amman University, Amman 19328, Jordan
  • Arwa Al-Khatib Pharmacological and Diagnostic Research Center (PDRC), Faculty of Pharmacy, Al-Ahliyya Amman University, Amman 19328, Jordan
  • Alia S. Khwaldeh Department of Medical Laboratory Sciences, Faculty of Pharmacy, Jadara University, Irbid, Jordan


Flavonoids, BCAAs, Antiglycation, Cytotoxicity, Co-incubation


Flavonoids and branched chain amino acids (BCAAs) have been recognized as attractive agents for research due to their role in diabetes and cancer. The present study investigates the antiglycation effect of BCAAs and flavonoid compounds and their cytotoxic effect on different colorectal cancer cell lines individually and in combination. This study was conducted by using RAW 264.7 cell line (murine monocyte-macrophage cell and Methylglyoxal (MGO) for inducing the glycation effect. The Sulforhodamine B (SRB) assay was used to study the cytotoxicity of BCAAs and flavonoids compounds using colorectal cell lines SW480, SW620 and CACO2. Flavonoids showed significant antiglycation effect compared to Aminoguanidine (AMG), while none of the BCAAs compounds displayed antiglycation effect. All flavonoids and branched chain amino acids showed less degree of cytotoxicity against SW620, CACO2 and SW480 as compared to cisplatin. The synergetic effects was only shown against SW620 and CACO2 cell lines by using the combination of Apigenin and BCAAs while antagonistic effect appeared when combination of Quercetin and Hesperidin were used against SW480, SW620 and CACO2 cell lines. Flavonoids such as Quercetin, Apigenin and Hesperidin could be used as a remedy for diabetes due to their surpassing effect upon antiglycation action of AMG. 


Aldini G, Vistoli G, Stefek M, Chondrogianni N, Grune T, Sereikaite J, Sadowska-Bartosz I, Bartosz G. Molecular strategies to prevent, inhibit, and degrade advanced glycoxidation and advanced lipoxidation end products. Free Rad Res. 2013; 47(1):93-137.

Chaudhuri J, Bains Y, Guha S, Kahn A, Hall D, Bose N, Gugliucci A, Kapahi P. The role of advanced glycation end products in aging and metabolic diseases: bridging association and causality. Cell Metab. 2018; 28(3):337-352.

Egaña-Gorroño L, López-Díez R, Yepuri G, Ramirez LS, Reverdatto S, Gugger PF, Shekhtman A, Ramasamy R, Schmidt AM. Receptor for Advanced Glycation End Products (RAGE) and Mechanisms and Therapeutic Opportunities in Diabetes and Cardiovascular Disease: Insights From Human Subjects and Animal Models. FrontCard Med. 2020; 7:37.

Rodriguez‐Teja M, Gronau JH, Breit C, Zhang YZ, Minamidate A, Caley MP, McCarthy A, Cox TR, Erler JT, Gaughan L, Darby S. AGE‐modified basement membrane cooperates with Endo180 to promote epithelial cell invasiveness and decrease prostate cancer survival. J Pathol. 2015; 235(4):581-592.

Peyroux J and Sternberg M. Advanced glycation endproducts (AGEs): pharmacological inhibition in diabetes. Pathol Biol. 2006; 54(7):405-419.

Alqaraleh M and Kasabri V. The Antiglycation Effect of Monomethyl Branched Chained Fatty Acid and Phytochemical Compounds and their Synergistic Effect on Obesity Related Colorectal Cancer Cell Panel. Rom J Diabet Nutr Metab Dis. 2019; 26(4):361-369.

Alqaraleh M, Kasabri V, Farha RA, Naffa RG, Yousef I, Aljaafreh A. Branched amino acids as potential biomarkers

in metabolic syndrome patients and as hypolipidemic compounds. EurAsian J BioSci. 2019; 13(2):2233-2241.

Alqaraleh M, Kasabri V, Mashallah S. Evaluation of Anticancer and Anti-Inflammatory Properties of Branched Chain Amino Acids. J Biochem Cell Bio. 2018; 1(2):108.

Alqaraleh M, Kasabri V, Al-Othman N. Evaluation of Pancreatic and Extra Pancreatic Effects of Branched Amino Acids. Rom J Diabet Nutr Metab Dis. 2019; 26(2):199-209.

Fernstrom JD. Branched-chain amino acids and brain function. The J Nutr. 2005; 135(6):1539-1546.

Ikeda T, Aizawa J, Nagasawa H, Gomi I, Kugota H, Nanjo K, Jinno T, Masuda T, Morita S. Effects and feasibility of exercise therapy combined with branched-chain amino acid supplementation on muscle strengthening in frail and prefrail elderly people requiring long-term care: a crossover trial. Appl Physiol Nutr Metab. 2016; 41(4):438-445.

Chen T, Ni Y, Ma X, Bao Y, Liu J, Huang F, Hu C, Xie G, Zhao A, Jia W, Jia W. Branched-chain and aromatic amino acid profiles and diabetes risk in Chinese populations. SciRep. 2016; 6:20594.

Nie C, He T, Zhang W, Zhang G, Ma X. Branched chain amino acids: beyond nutrition metabolism. Inter J Mol Sci. 2018; 19(4):954.

Hagiwara A, Nishiyama M, Ishizaki S. Branched‐chain amino acids prevent insulin‐induced hepatic tumor cell proliferation by inducing apoptosis through mTORC1 and mTORC2‐dependent mechanisms. J Cellular Physiol. 2012;


Khleifat KM, Abboud MM, Al-Mustafa AH, Al-Sharafa KY. Effects of carbon source and Vitreoscilla hemoglobin (VHb) on the production of β-galactosidase in Enterobacter aerogenes. Curr Microbiol. 2006; 53(4):277.

Abboud MM, Aljundi IH, Khleifat KM, Dmour S. Biodegradation kinetics and modeling of whey lactose by bacterial hemoglobin VHb-expressing Escherichia colistrain. Biochem Eng J. 2010; 48(2):166-172.

Althunibat OY, Qaralleh H, Al-Dalin SY, Abboud M, Khleifat K, Majali IS, Aldal'in HK, Rayyan WA, Jaafraa A. Effect of thymol and carvacrol, the major components of Thymus capitatus on the growth of Pseudomonas aeruginosa. J Pure Appl Microbiol. 2016; 10(1):367-374.

Al-Tawarah NM, Qaralleh H, Khlaifat AM, Nofal MN, Alqaraleh M, Khleifat KM, Al-limoun MO, Al Shhab MA. Anticancer and Antibacterial Properties of Verthemia Iphionides Essential Oil/Silver Nanoparticles. Biomed Pharm J. 2020; 3(3):1175-1185.

Tungmunnithum D, Thongboonyou A, Pholboon A, Yangsabai A. Flavonoids and other phenolic compounds from medicinal plants for pharmaceutical and medical aspects: An overview. Med. 2018; 5(3):93.

Wondrak GT, Cervantes-Laurean D, Roberts MJ, Qasem JG, Kim M, Jacobson EL, Jacobson MK. Identification of α-dicarbonyl scavengers for cellular protection against carbonyl stress. Biochem Pharmacol. 2002; 63(3):361-373.

Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, Minor L. Cell viability assays assay guidance manual. Assay Guidance Manual. 2004; 1-23.

Vichai V and Kirtikara K. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat Protoc. 2006;1(3):1112.

Marks LS, Hess DL, Dorey FJ, Macairan ML, Santos PBC, Tyler VE. Tissue effects of saw palmetto and finasteride:

use of biopsy cores for in situ quantification of prostatic androgens. Urol 2001; 57(5):999-1005.

Thornalley PJ, Langborg A, Minhas HS. Formation of glyoxal, methylglyoxal and 3-deoxyglucosone in the glycation of proteins by glucose. Biochem J. 1999;344(1):109-116.

Wu C-H and Yen G-C. Inhibitory effect of naturally occurring flavonoids on the formation of advanced glycation endproducts. J Agric Food Chem. 2005;53(8):3167-3173.

Boussahel S, Cacciola F, Dahamna S, Mondello L, Saija A, Cimino F, Speciale A, Cristani M. Flavonoid profile, antioxidant and antiglycation properties of Retama sphaerocarpa fruits extracts. Nat Prod Res. 2018;32(16):1911-1919.

Crascì L, Lauro MR, Puglisi G, Panico A. Natural antioxidant polyphenols on inflammation management: Anti-glycation activity vs metalloproteinases inhibition. Critic Rev Food Sci Nutr. 2018; 58(6):893-904.

Mazumder MAR and Hongsprabhas P. Genistein as antioxidant and antibrowning agents in vivo and in vitro: A

review. Biomed Pharmacother. 2016; 82:379-392.

Silvan JM, Srey C, Ames JM, del Castillo MD. Glycation is regulated by isoflavones. Food Funct. 2014; 5(9):2036-2042.

Zhenyukh O, Civantos E, Ruiz-Ortega M, Sánchez MS, Vázquez C, Peiró C, Egido J, Mas S. High concentration of

branched-chain amino acids promotes oxidative stress, inflammation and migration of human peripheral blood mononuclear cells via mTORC1 activation. Free Rad Bio Med. 2017; 104:165-177.

Zhang W, Huang Q, Zeng Z, Wu J, Zhang Y, Chen Z. Sirt1 inhibits oxidative stress in vascular endothelial cells. Oxid Med Cell Longev. 2017; 2017.

Havsteen BH. The biochemistry and medical significance of the flavonoids. Pharmacol Ther. 2002; 96(2-3):67-202.

Abotaleb M, Samuel SM, Varghese E, Varghese S, Kubatka P, Liskova A, Büsselberg D. Flavonoids in cancer and

apoptosis. Cancers. 2019; 11(1):28.

Sahpazidou D, Geromichalos GD, Stagos D, Apostolou A, Haroutounian SA, Tsatsakis AM, Tzanakakis GN, Hayes

AW, Kouretas D. Anticarcinogenic activity of polyphenolic extracts from grape stems against breast, colon, renal and

thyroid cancer cells. Toxicol Lett. 2014; 230(2):218-224.

Vargas AJ and Burd R. Hormesis and synergy: pathways and mechanisms of quercetin in cancer prevention and

management. Nutr Rev. 2010; 68(7):418-428.

Shen X, Si Y, Wang Z, Wang J, Guo Y, Zhang X. Quercetin inhibits the growth of human gastric cancer stem cells by inducing mitochondrial-dependent apoptosis through the inhibition of PI3K/Akt signaling. Inter J Mol Med. 2016; 38(2):619-626.

Yan X, Qi M, Li P, Zhan Y, Shao H. Apigenin in cancer therapy: anti-cancer effects and mechanisms of action. Cell Biosci. 2017; 7(1):50.

Devi KP, Rajavel T, Nabavi SF, Setzer WN, Ahmadi A, Mansouri K, Nabavi SM. Hesperidin: A promising anticancer agent from nature. Ind Crops Prod. 2015;76:582-589.

Wubetu GY, Utsunomiya T, Ishikawa D, Ikemoto T, Yamada S, Morine Y, Iwahashi S, Saito Y, Arakawa Y, Imura S, Arimochi H. Branched chain amino acid suppressed insulin-initiated proliferation of human cancer cells through induction of autophagy. Anticancer Res. 2014;34(9):4789-4796.




How to Cite

Alqaraleh, M., Kasabri, V., Aljaafreh, A., Al-Majali, I., Al-Othman, N., Khleifat, K., … Khwaldeh, A. S. (2021). Evaluation of the Antiglycation Effect of Branched Chain Amino Acids and Phytochemical Compounds on RAW 264.7 Cell Line and their Synergistic Effect on Colorectal Cancer Cell Line Panel: doi.org/10.26538/tjnpr/v5i1.11. Tropical Journal of Natural Product Research (TJNPR), 5(1), 88–93. Retrieved from https://tjnpr.org/index.php/home/article/view/827

Most read articles by the same author(s)