Effect of ethanol extracts from Cnidoscolus aconitifolius leaves on foam cell formation and effect of leaf age and harvesting season on phytochemicals, antioxidant, cytotoxicity and nitric oxide production

Main Article Content

Niramai Fangkrathok
Bung-on Prajanban


Cnidoscolus aconitifolius or Chaya (Euphorbiaceae) leaf has been used as a food and a medicinal plant for treatment of hypertension. Antihypercholesterolemic and antihypertriglyceridemic activities of C. aconitifolius leaf extract have been reported. It is possible that Chaya leaf may have an inhibitory effect on foam cell formation. Foam cells are involved with an atherosclerosis that related with high level of LDL cholesterol and hypertension. However, harvesting season and age of leaf may affect to phytochemicals in leaves that may relate to its biological activity. Therefore, aims of this study were to investigate the effects of C. aconitifolius ethanolic extracts (CAE) on foam cell formation, nitric oxide (NO) and TNF-α. The differences in phytochemicals influencing from harvesting season and age stage of its leaves were also studied. Methods: Total phenolic and flavonoid contents were studied by using Folin-Ciocalteu and aluminum chloride reaction, respectively. Amount of kaempferol was analysed using HPLC. Antioxidative activity was determined by using DPPH and FRAP assay. In RAW264.7 cells, cytotoxicity, NO and TNF-α production were studied by using MTT assay, Griess reaction and ELISA, respectively. Foam cell formation was determined by using Oil Red O staining. Results: The difference in leaf age and harvesting season affected to its phytochemicals and antioxidative activity. Leaves that were harvested in dry season showed higher antioxidative activity, total phenolic and total flavonoid contents than those of leaves that were harvested in rainy season. Aging leaf extracts showed lower cytotoxicity than young and mature leaf extracts. All extracts exhibited inhibitory activity on NO production in LPS-induced RAW264.7 cells. Extracts that were harvested in dry season showed greater inhibitory activity on NO production than those of rainy season. Mature leaf extract harvested in dry season showed higher inhibitory effect on NO production (IC50 of 66.16 ± 1.13 µg/mL) than those of the others. Co-treatment between mature leaf extract harvested in dry season and oxLDL could lower lipid accumulation and TNF-α production in foam cells than those of oxLDL treatment alone. These results indicate that CAE, especially mature leaf extract harvested in dry season, may reduce lipid accumulation in oxLDL-inducing foam cells via suppression of NO and TNF-α production. Conclusion: Mature leaf extract of C. aconitifolius that harvested in dry season had high antioxidants, low cytotoxicity, inhibitory activities on NO production, foam cell formation and TNF-α production. Therefore, C. aconitifolius leaf extract can be an alternative choice for health beneficial product development.


Download data is not yet available.

Article Details

Pharmaceutical Sciences


Achi NK, Ohaeri OC, Ijeh II, Eleazu C. Modulation of the lipid profile and insulin levels of streptozotocin induced diabetic rats by ethanol extract of Cnidoscolus aconitifolius leaves and some fractions: Effect on the oral glucose tolerance of normoglycemic rats. Biomed Pharmacother 2017; 86: 562-569.

Agamou JAA, Fombang EN, Mbofung CMF. Particular benefits can be attributed to Moringa oleifera Lam leaves based on origin and stage of maturity. JEBAS 2015; 3(6): 541-555.

Ajiboye BO, Ojo OA, Okesola MA, Oyinloye BE, Kappo AP. Ethyl acetate leaf fraction of Cnidoscolus aconitifolius (Mill.) I. M. Johnst: antioxidant potential, inhibitory activities of key enzymes on carbohydrate metabolism, cholinergic, monoaminergic, purinergic, and chemical fingerprinting. Int J Food Prop 2018; 21(1): 1697–1715.

Akomolafe SF, Oboh G, Oyeleye SI, Boligon AA. Aqueous extract from Ficus capensis leaves inhibits key enzymes linked to erectile dysfunction and prevent oxidative stress in rats’ penile tissue. NFS J 2016; 4: 15–21.

Ares MPS, Kallin B, Eriksson P, Nilsson J. Oxidized LDL induces transcription factor AP-1 but inhibits activation of NF-κB in human vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 1995; 15: 1584-1590.

Baillie AG, Coburn CT, Abumrad NA. Reversible binding of long-chain fatty acids to purified FAT, the adipose CD36 homolog. J Membr Biol 1996; 153: 75–81.

Daniels CW, Rautenbach F, Marnewick JL, Valentine AJ, Babajide OJ, Mabusela WT. Environmental stress effect on the phytochemistry and antioxidant activity of a South African bulbous geophyte, Gethyllis multifolia L. Bolus. S Afr J Bot 2015; 96: 29-36.

de Lima TM, de Sa Lima L, Scavone C, Curi R. Fatty acid control of nitric oxide production by macrophages. FEBS Letters 2006; 580(13): 3287-3295.

Dian-Nashiela F, Noriham A, Nooraain H, Azizah AH. Antioxidant activity of herbal tea prepared from Cosmos caudatus leaves at different maturity stages. IFRJ 2015; 22(3): 1189–1194.

Jay AG, Chen AN, Paz MA, Hung JP, Hamilton JA. CD36 binds oxidized low density lipoprotein (LDL) in a mechanism dependent upon fatty acid binding. J Biol Chem 2015; 290(8): 4590–4603.

Jiménez-Arellanes MA, García-Martínez I, Rojas-Tomé S. Potencial biológico de especies medicinales del género Cnidoscolus (Euphorbiacea). Rev Mex Ciencias Farm 2014; 45(4): 1-6.

Jovinge S, Ares MPS, Kallin B, Nilsson J. Human Monocytes/Macrophages Release TNF-α in Response to Ox-LDL. Arterioscler Thromb Vasc Biol 1996; 16: 1573–1579.

Krieger M. Scavenger receptor class B type I is a multiligand HDL receptor that influences diverse physiologic systems. J Clin Invest 2001; 108: 793–797.

Kumar A, Or-Rashid MM, Alzahol O, McBride B. Fatty acid profile of chaya (Cnidoscolus aconitifolius) leaves and fodder. Indian J Anim Sciences 2011; 81(8): 95-100.

Loarca-Piña G, Mendoza S, Ramos-Gómez M, Reynoso R. Antioxidant, antimutagenic, antidiabetic activities of edible leaves from Cnidoscolus chayamansa Mc. Vaugh. J Food Sci 2010; 75(2): 68-72.

Maiolino G, Rossitto G, Caielli P, Bisogni V, Rossi GP, Calò LA. The role of oxidized low-density lipoproteins in atherosclerosis: The myths and the facts. Mediators Inflamm 2013; 2013: 714653.

Miranda-Velasquez L, Oranday-Cardenas A, Lozano-Garza H, RivasMorales C, ChamorroCevallos G, Cruz-Vega DE. Hypocholesterolemic activity from the leaf extracts of Cnidoscolus chayamansa. Plant Food Hum Nutr 2010; 65: 392-395.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65(1-2): 55-63.

Ncube B, Finnie JF, van Staden J. Seasonal variation in antimicrobial and phytochemical properties of frequently used medicinal bulbous plants from South Africa. S Afr J Bot 2011; 77(2): 387-396.

Nguyen MT, Fernando S, Schwarz N, Tan JTM, Bursill CA, Psaltis PJ. Inflammation as a therapeutic target in atherosclerosis. J Clin Med 2019; 8: 1109.

Nobossé P, Fombang EN, Mbofung CMF. Effects of age and extraction solvent on phytochemical content and antioxidant activity of fresh Moringa oleifera L. leaves. Food Sci Nutr 2018; 6(8): 2188-2198.

Olaniyan M, Ozuaruoke D, Afolabi T. Cholesterol lowering effect of Cnidoscolus aconitifolius leave extracts in egg yolk induced hypercholesterolemia in rabbit. J Adv Med Med Res 2017; 23(1): 1-6.

Onasanwo SA, Oyagbemi AA, Saba AB. Antiinflammatory and analgesic properties of the ethanolic extract of Cnidoscolus aconitifolius in rats and mice. J Basic Clin Physiol Pharmacol 2011; 22(1-2): 34-41.

Orji OU, Ibiam UA, Aja PM, Ugwu Okechukwu PC, Uraku AJ, Aloke C, et al. Evaluation of the phytochemical and nutritional profiles of Cnidoscolus aconitifolius leaf collected in Abakaliki South East Nigeria. WJMS 2016; 13(3): 213-217.

Oyagbemi AA, Odetola AA. Hepatoprotective and nephroprotective effects of Cnidoscolus aconitifolius in protein energy malnutrition induced liver and kidney damage. Pharmacognosy Res 2013; 5(4): 260-264.

Park YM. CD36, a scavenger receptor implicated in atherosclerosis. Exp Mol Med 2014; 46(6): e99.

Pillai KK, Chidambaranathan N, Mohamed Halith M, Jayaprakash S, Narayanan N. Anti-hyperglycemic effect of alcoholic extracts of Cnidoscolus chayamansa in experimental diabetes and their effects on key metabolic enzymes involved in carbohydrate metabolism. IJRPC 2012; 2(1): 179-187.

Ross R. Atherosclerosis--an inflammatory disease. N Engl J Med 1999; 340(2): 115-126.

Roy DN, Ferdiousi N, Khatun T, Moral RA. Phytochemical screening, nutritional profile and anti-diabetic effect of ethanolic leaf extract of Cnidoscolus aconitifolius in streptozotocin induce diabetic mice. IJBCP 2016; 5(5): 2244-2250.

Samuel I, Arthur N, Jude E, Henrietta C. Antihyperglycaemic efficacy of Cnidoscolus aconitifolius compared with glibenclamide in alloxan-induced diabetic Wistar rats. Int Res J Medical Sci 2014; 2(3): 1-4.

Schreck R, Rieber P, Baeuerle PA. Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-κB transcription factor and HIV-1. EMBO J 1991; 10: 2247-2258.

Shi H, Kokoeva MV, Inouye K, Tzameli I, Yin H, Flier JS. TLR4 links innate immunity and fatty acid-induced insulin resistance. J Clin Invest 2006; 116: 3015–3025.

Sreelatha S, Padma PR. Antioxidant activity and total phenolic content of Moringa oleifera leaves in two stages of maturity. Plant Food Hum Nutr 2009; 64(4): 303-311.

Su X, Abumrad NA. Cellular fatty acid uptake: a pathway under construction. Trends Endocrinol Metab 2009; 20: 72–77.

Takashiba S, Shapira L, Amar S, Van Dyke TE. Cloning and characterization of human TNFα promoter region. Gene 1993; 131: 307-308.

Zhang H, Zhai Z, Zhou H, Li Y, Li X, Lin Y, et al. Puerarin inhibits oxLDL-induced macrophage activation and foam cell formation in human THP1 macrophage. BioMed Res Int 2015; 2015(403616): 1-8.