Protective role of glycine and kolaviron on lipopolysaccharide-induced alterations of raw U937 cells and U937-derived macrophages


  • Tebekeme OKOKO Niger Delta University, Department of Biochemistry (NG)
  • Solomon A. NDONI Niger Delta University, Department of Biochemistry (NG)



cell viability, glycine, kolaviron, lipopolysaccharide, macrophages


The effect of glycine and kolaviron on lipopolysaccharide-induced U937 cell damage and activation of U937-derived macrophages was studied. U937 cells were incubated with either glycine or kolaviron or both for 24 h before exposure to lipopolysaccharide. Cell viability and production of reactive oxygen species (ROS) were later assessed. In the other experiment, the U937 cells were transformed to the macrophage form using phorbol 12-myristate 13-acetate and incubated with or without glycine or kolaviron or both before exposure to lipopolysaccharide. Production of TNF-α, IL-1, IL-6 and NO were later assessed. The expression of the antioxidant enzymes- superoxide dismutase (SOD) and catalase (CAT) was also evaluated via reverse transcription polymerase chain reaction (RT PCR). It revealed that lipopolysaccharide caused significant cell death and production of reactive oxygen species that was reduced by glycine and kolaviron. Glycine and kolaviron also reduced lipopolysaccharide-mediated secretion of TNF-α, IL-1, IL-6 and NO in U937-derived macrophages. In some cases, pre-incubation of cells with both glycine and kolaviron was better than the individual responses. Glycine and kolaviron also reduced lipopolysaccharide-induced alterations in the expression of SOD and CAT (p<0.05). The study shows that both glycine and kolaviron (either separately or in combination) reduced lipopolysaccharide-mediated alterations in U937 cells and U937-derived macrophages.


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Abarikwu SO (2015). Anti-inflammatory effects of kolaviron modulate the expressions of inflammatory marker genes, inhibit transcription factors ERK1/2, p-JNK, NF-κB, and activate Akt expressions in the 93RS2 Sertoli cell lines. Molecular and Cellular Biochemistry 401:197-208.

Adaramoye OA, Lawal SO (2015). Kolaviron, a biflavonoid fraction from Garcinia kola, protects against isoproterenol-induced injury by mitigating cardiac dysfunction and oxidative stress in rats. Journal of Basic Clinical Physiology and Pharmacology 26:65-72. https://doi/10.1515/jbcpp-2013-0139

Akinmoladun AC, Akinrinola BL, Olaleye MT, Farombi EO (2015). Kolaviron, a Garcinia kola biflavonoid complex, protects against ischemia/reperfusion injury: pertinent mechanistic insights from biochemical and physical evaluations in rat brain. Neurochemical Research 40:777-787.

Aldrich MB, Sevick-Muraca EM (2013). Cytokines are systemic effectors of lymphatic function in acute inflammation. Cytokine 64:362-369.

Angeloni C, Spencer JP, Leonani E, Biagi PL, Hrelia S (2007). Role of quercetin and its in vivo metabolites in protecting H9c2 cells against oxidative stress. Biochimie 89:73-82.

Apalowo OE, Musa SE, Asaolu F, Apata JT, Oyedeji T, Babalola OO (2018). Protective roles of kolaviron extract from Garcinia kola seeds against isoniazid-induced kidney damage in Wistar rats. European Journal of Medicinal Plants 24:1-8.

Chen TJ, Jeng JY, Lin CW, Wu CY, Chen YC (2006). Quercetin inhibition of ROS-dependent and-independent apoptosis in rat glioma C6 cells. Toxicology 223:113-126.

Chow JM, Shen SC, Huan SK, Lin HY, Chen YC (2005). Quercetin, but not rutin and quercitrin, prevention of H2O2-induced apoptosis via anti-oxidant activity and heme oxygenase 1 gene expression in macrophages. Biochemical Pharmacology 69:1839-1851.

Dou Y, Wang X, Yu G, Wang S, Tian M, Qi J, … Yu S (2017). Disruption of the M949_RS01915 gene changed the bacterial lipopolysaccharide pattern, pathogenicity and gene expression of Riemerella anatipestifer. Veterinary Research 48(1):6.

Esiegwu AC, Okoli IC, Emenalom OO, Esonu BO, Udedibie ABI (2014). The emerging benefits of the African wonder nut (Garcinia kola Heckel): A review. Global Journal of Animal Science Research 2:170-183.

Förstermann U, Xia N, Li H (2017). Roles of vascular oxidative stress and nitric oxide in the pathogenesis of atherosclerosis. Circulation Research 120:713-735.

Franceschelli S, Pesce M, Ferrone A, De Lutiis MA, Patruno A, Grilli A, … Speranza L (2014). Astaxanthin treatment confers protection against oxidative stress in U937 cells stimulated with lipopolysaccharide reducing O2− production. PLoS One 9:e88359. https://doi:10.1371/journal.pone.0088359

Hsieh YH, Kuo PM, Chien SC, Shyur LF, Wang SY (2007). Effects of Chamaecyparis formosensis Matasumura extractivities on lipopolysaccharide-induced release of nitric oxide. Phytomedicine 14:675-680.

Huang Q, Hu XL (2017). Effects of salidroside on the secretion of inflammatory mediators induced by lipopolysaccharide in murine macrophage cell line J774. Acta Physiologica Sinica 69:41-46.

Huang Z, Kraus VB (2016). Does lipopolysaccharide-mediated inflammation have a role in OA? Nature Reviews Rheumatology 12:123.

Hwang BY, Lee JH, Koo TH, Hong YS, Ro IS, Lee KS, Lee JJ (2002). Furanoligularenone, an eremophilane from Ligularia fischerri inhibits the lipopolysaccharide-induced production of nitric oxide and prostaglandin E2 in macrophage RAW264.7 cells. Planta Medica 68:101-105.

Hwang PA, Chien SY, Chan YL, Lu MK, Wu CH, Kong ZL, Wu CJ (2011). Inhibition of lipopolysaccharide (LPS)-induced inflammatory responses by Sargassum hemiphyllum sulfated polysaccharide extract in RAW 264.7 macrophage cells. Journal of Agricultural and Food Chemistry 59:2062-2068.

Iwu M, Okunji C, Tchimene M, Anele N, Chah K, Akpa PA, Onunkwo GC (2009). Stability of cough linctus (streptol) formulated from medicinal plant extracts. Chemical and Pharmaceutical Bulletin 57:229-232.

Iwu MM (1985). Antihepatoxic constituents of Garcinia kola seeds. Experientia 41:699-700.

Jain M, Nilsson R, Sharma S, Madhhusudhan N, Kitami T, Souza AL, … Mootha VK (2012). Metabolic profiling identifies a key role for glycine in rapid cancer cell proliferation. Science 336:1040-1044.

Koga T, Meydani M (2001). Effect of plasma metabolites of (+)-catechin and quercetin on monocyte adhesion to human aortic endothelial cells. American Journal of Clinical Nutrition 73:941-948.

Laposata M (2019). Laboratory Medicine: Diagnosis of Disease in Clinical Laboratory. 3rd ed. New York. McGraw-Hill Education.

Lee SY, Cho JY (2009). Inhibitory effects of honokiol on LPS and PMA-induced cellular responses of macrophages and monocytes. BMB Reports 42:574-579.

Li P, Wu G (2018). Roles of dietary glycine, proline, and hydroxyproline in collagen synthesis and animal growth. Amino Acids 50:29-38.

Liu T, Li J, Liu Y, Xiao N, Suo H, Xie K, Yang C, Wu C (2012). Short-chain fatty acids suppress lipopolysaccharide-induced production of nitric oxide and proinflammatory cytokines through inhibition of NF-κB pathway in RAW264.7 cells. Inflammation 35:1676-1684.

Liu X, Lu J, Liao Y, Liu S, Chen Y, He R, Men L, Lu C, Chen Z, Li S, Xiong G (2019). Dihydroartemisinin attenuates lipopolysaccharide-induced acute kidney injury by inhibiting inflammation and oxidative stress. Biomedicine and Pharmacotherapy 117:109070.

Modlinger PS, Wilcox CS, Aslam S (2004). Nitric oxide, oxidative stress, and progression of chronic renal failure. Seminars in Nephrology 24:354-365.

Nishimura Y, Lemasters JJ (2001). Glycine blocks opening of a death channel in cultured hepatic sinusoidal endothelial cells during chemical hypoxia. Cell Death and Differentiation 8:850-858.

Okoko T, Oruambo IF (2009). Inhibitory activity of quercetin and its metabolite on lipopolysaccharide-induced activation of macrophage U937 cells. Food and Chemical Toxicology 47:809-812.

Okoko T (2018). Kolaviron and selenium reduce hydrogen peroxide-induced alterations of the inflammatory response. Journal of Genetic Engineering and Biotechnology 16:485-490.

Okuda S, Sherman DJ, Silhavy TJ, Ruiz N, Kahne D (2016). Lipopolysaccharide transport and assembly at the outer membrane: the PEZ model. Nature Reviews Microbiology 14:337-345.

Oyagbemi AA, Bester D, Esterhuyse J, Farombi EO (2017). Kolaviron, a biflavonoid of Garcinia kola seed mitigates ischemic/reperfusion injury by modulation of pro-survival and apoptotic signaling pathways. Journal of Intercultural Ethnopharmacology 6:42-49. https://doi/10.5455/jice.20160923100223

Qiao S, Luo Q, Zhao Y, Zhang XC, Huang Y (2014). Structural basis for lipopolysaccharide insertion in the bacterial outer membrane. Nature 511:108-111.

Razak MA, Begum PS, Viswanath B, Rajagopal S (2017). Multifarious beneficial effect of nonessential amino acid, glycine: a review. Oxidative Medicine and Cellular Longevity 1716701.

Rhee SH (2014). Lipopolysaccharide: Basic biochemistry, intracellular signaling, and physiological impacts in the gut. Intestinal Research 12:90-95.

Rios ECS, de Lima TM, Moretti AIS, Soriano FG (2016). The role of nitric oxide in the epigenetic regulation of THP-1 induced by lipopolysaccharide. Life Science 147:110-116.

Ruiz-Maena M, Puna P, Garcia-Dorado D, Rodriguez-Sinovas A, Barba I, Miro-Casas E, … Soler-Soler J (2004). Glycine protects cardiomyocytes against lethal reoxygenation injury by inhibiting mitochondrial permeability transition. Journal of Physiology 558:873-882.

Ryu SJ, Choi J, Lee JS, Choi HS, Yoon KY, Hwang JH, Kim KJ, Lee BY (2018). Compound K inhibits the lipopolysaccharide-induced inflammatory responses in Raw 264.7 Cell Line and Zebrafish. Applied Science 8:924.

Shafiekhani M, Ommati MM, Azarpira N, Heidari R, Salarian AA (2019). Glycine supplementation mitigates lead-induced renal injury in mice. Journal of Experimental Pharmacology 11:15-22.

Speranza L, Franceschelli S, Pesce M, Reale M, Menghini L, Vinciguerra I, De Lutiis MA, Felaco M, Grilli A (2010). Anti-inflammatory effects in THP‐1 cells treated with verbascoside. Phytotherapy Research 24:1398-1404.

Tang X, Liu B, Wang X, Yu Q, Fang R (2018). Epidermal growth factor, through alleviating oxidative stress, protect IPEC-J2 cells from lipopolysaccharides-induced apoptosis. International Journal of Molecular Sciences 19:848.

Wang Y, Wang L, Wise JT, Shi X, Chen Z (2020). Verteporfin inhibits lipopolysaccharide-induced inflammation by multiple functions in RAW 264.7 cells. Toxicology and Applied Pharmacology 387:114852.

Wang Z, Zhang J, Chen L, Li J, Zhang H, Guo X (2019). Glycine suppresses AGE/RAGE signaling pathway and subsequent oxidative stress by restoring Glo1 function in the aorta of diabetic rats and in HUVECs. Oxidative Medicine and Cellular Longevity 2019:4628962.

Weinberg JM, Bienholz A, Venkatachalam MA (2016). The role of glycine in regulated cell death. Cellular and Molecular Life Sciences 73:2285-3308.

Wheeler MD, Ikejema K, Enomoto N, Stacklewitz RF, Seabra V, Zhong Z, Yin M, Schemmer P, Rose ML, Rusyn I, Bradford B, Thurman RG (1999). Glycine: a novel anti-inflammatory immune-nutrient. Cellular and Molecular Life Sciences 56:843-856.

Ye J, Guan M, Lu Y, Zhang D, Li C, Zhou C (2019). Arbutin attenuates LPS-induced lung injury via Sirt1/Nrf2/NF-κBp65 pathway. Pulmonary Pharmacology and Therapeutics 54:53-59.

Yu GM, Tan W (2019). Melatonin inhibits lipopolysaccharide-induced inflammation and oxidative stress in cultured mouse mammary tissue. Mediators of Inflammation 2019:8597159.

Zhang H, Peng A, Yu Y, Guo S, Wang M, Wang H (2019). L-arginine protects ovine intestinal epithelial cells from lipopolysaccharide-induced apoptosis through alleviating oxidative stress. Journal of Agricultural and Food Chemistry 67(6):1683-1690.

Zhou Q, Xie H, Zhang L, Stewart JK, Gu X-X, Ryan JJ (2006). cis-Terpenones as an effective chemoprotective agent against aflatoxin-B1-induced cytotoxicity and TCDD-induced P450 1A/B activity in HepG2 cells. Chemical Research in Toxicology 19:1415-1419.



How to Cite

OKOKO, T., & NDONI, S. A. (2022). Protective role of glycine and kolaviron on lipopolysaccharide-induced alterations of raw U937 cells and U937-derived macrophages. Notulae Scientia Biologicae, 14(1), 11082.



Research articles
DOI: 10.15835/nsb14111082