Isolation and characterization of the antibreast carcinoma cell growth components of Vernonia amygdalina extracts

Abstract

Vernonia amygdalina (VA) is widely used for medicinal and food purposes in tropical Africa. Many health benefits (antioxidant, antimicrobial, anticancer activities and more) of VA extracts have been reported. The mechanisms of actions have also been described. We have previously reported that VA extracts elicited growth inhibitory activities in human estrogen receptor-positive (ER(+)) cells (MCF-7 cells) and ductal carcinoma cells (BT-549) in vitro. The active components in the organic solvent (chloroform)-extracted VA have been previously determined. However, the active components in the ethanolic extracts of VA have not been previously studied. Hence, the objectives of this study are to isolate and characterize the active components of the ethanolic extracts of VA using liquid-liquid extraction, thin layer chromatography and column techniques. Fractionation of the ethanolic extracts of VA yielded three fractions named A1, A2 and A3, and A2 retained the DNA synthesis-inhibitory activity of the extracts. Subsequent fractionation of A2 yielded fraction A2B whose activity was 16 and three times more potent than the ethanolic fraction and fraction A2, respectively. The treatment of cells with 100 μg/mL of either the ethanolic VA extracts, fraction A2 or fraction A2B resulted in a 23% (P< 0.01), 86% (P < 0.0001) and 97% (P < 0.0001) inhibition of DNA synthesis compared with vehicle-treated controls, respectively. Further purification of A2B by high-speed countercurrent chromatography and confirmed by spectroscopic analysis revealed that the major active components of A2B (65% by weight) were steroid glucosides.

Steroidal Saponins from Vernonia amygdalina Del. and Their Biological Activity

“…Over the years, several studies on the chemical components of this species, including flavonoids [ 2 ], sesquiterpene lactones [ 3 ], steroidal saponins [ 4 , 5 , 6 ], and fatty acids [ 7 ], have been performed. Previous studies have indicated different bioactivities of this species, including anti-inflammation [ 8 ], anti-malaria [ 9 ], anti-obesity [ 10 , 11 ], antioxidant [ 12 , 13 ], anti-tumor [ 14 ], and other activities [ 15 ]. Water and chloroform extracts of V. amygdalina (VA) interfere with the DNA synthesis of MCF-7 cells in breast cancer, affecting the activity of ERKs in vitro and inhibiting human breast cancer cells [ 16 , 17 ].…”

“…Via the MTT method, it has been verified that MCF-7 cells are inhibited in vitro and the DNA synthesis of BT-549 cells in breast cancer is disturbed by the addition of VA extract [ 18 , 19 ]. Moreover, three active fractions extracted from ethanol extracts of V. amygdalina have an inhibiting effect on tumor cells, the essential component being steroidal saponins [ 14 ]. So far, a certain amount of steroidal saponins has been obtained from V. amygdalina; however, studies on the anti-tumor activity are rare.…”

Supplementation of broiler feed with leaves of Vernonia amygdalina and Azadirachta indica protected birds naturally infected with Eimeria sp.

“…Plants native to Nigeria have also been shown to possess anticoccidial activity (Nweze and Obiwulu, 2009). Nigeria herbal plants Vernonia amygdalina (VA) and Azadirachta indica (AI) have been considered potentially effective chemo-preventive and chemotherapeutic agents against breast cancer cells (Luo et al, 2010;Kumar et al, 2009). V. amygdalina was reported to have an anthelmintic and hepatoprotective properties (Adedapo et al, 2007;Adesanoye and Farombi, 2010).…”

Countercurrent Separation of Natural Products: An Update

“…Table S10, Supporting Information , summarizes the use of CCS to prepare fractions for bioassay screening of natural products in articles published since 2007. 20 , 35 , 41 , 44 , 74 , 94 , 102 , 105 , 116 , 122 , 128 , 135 – 137 , 139 , 229 , 230 , 279 , 284 , 285 , 295 , 298 , 307 , 308 , 324 – 326 , 344 , 345 , 359 – 402 …”