Phenolic Compounds from Water-Ethanol Extracts of Tetrapleura tetraptera Produced in Cameroon, as Potential Protectors against In Vivo CCl4-Induced Liver Injuries.

BACKGROUND: Liver diseases are a global health problem. Medicinal plants are being increasingly used to manage a wide variety of diseases including liver disorders. The aim of this study was to investigate the antioxidant properties and hepatoprotective activity of polyphenolic extract from the fruits of Tetrapleura tetraptera (T. tetraptera). RESULTS: The extract of T. tetraptera was administered at doses of 50 mg/kg and 100 mg/kg for 07 per os to rats before the induction of hepatotoxicity with of 2 ml/kg of 1:1 (v/v) carbon tetrachloride (CCl4) and olive oil through intraperitoneal route. The in vitro antioxidant and radical scavenging properties of T. tetraptera were conducted by the FRAP method, the phosphomolybdate method, and the inhibition potential of DPPH, ABTS, OH, and NO radicals. The extraction yield of T. tetraptera was 19.35%. This extract contains polyphenols (273.48 mg CAE/g DM), flavonoids (5.2549 mg SE/g DM), and flavonols (1.615 mg SE/g DM). This extract showed in vitro antioxidant activity, an inhibitor power of various free radicals, and radical scavenging potential dose-dependent. The fifty-percent inhibitory concentration of the extract (IC50) for the studied radical varied from 28.16 to 136 µg/L. In rats treated with the extract of T. tetraptera, in a dose-dependent manner, the levels of hepatotoxicity markers such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) significantly increased while the enzyme activities of superoxide dismutase (SOD), catalase (CAT), and the level of reduced glutathione (GHS) significantly increased compared to the control group. CONCLUSIONS: The extracts from the fruit of T. tetraptera demonstrate antioxidant activity and hepatoprotective effects.

In vitro organo-protective effect of bark extracts from Syzygium guineense var macrocarpum against ferric-nitrilotriacetate-induced stress in wistar rats homogenates.

BACKGROUND: Overconsumption of oxygen in mammalian cells often lead to the production of reactive oxygen species (ROS) resulting from different mechanisms. Escape of scavenging enzymes/components or nutritional failure are the most important origins. Plant-derived molecules may protect biological molecules either by quenching free radicals, delaying or preventing the ROS formation or by restoring antioxidant enzymes activities. The present study assessed the antioxidant, phenolic profile and protective effect of barks extracts of Syzyguim guineense var macrocarpum against ferric nitriloacetate-induced stress in the liver, heart kidney and brain tissues of wistar rat homogenates. METHODS: Three extracts (aqueous, ethanol and aqueous-ethanol) from the barks of S. guineense var macrocarpum were used in this study. The spectrophotometric standardized methods were used to determine the free radical scavenging and antioxidant potential of the extracts. The protective properties of these plant extracts were also investigated as well as the quantification of secondary metabolites content (total phenolic, flavonoids and flavonols content). The HPLC method helped for characterizing phenolic compounds present in these extracts. RESULTS AND DISCUSSION: All the extracts exhibited a free radical scavenging potential in a concentration dependent manner which varied from 15.18 ± 0.80 to 97.15 ± 0.71 % depending to the type of extract and the method used. The ethanol extract had the higher phenolic content (432.85 mg QE/g extract), including total flavonoids (961.66 mg QE/g extract) and flavonols content (25.12 mg QE/g extract) and higher total antioxidant capacity. Among the phenolic compounds present in the extracts, the HLPC profile revealed the presence of syringic acid and apigenin in all the extracts. The extracts demonstrated their protective effect mostly in liver and brain homogenates by delaying or preventing lipid peroxidation, restoring enzymatic activities and enhancing glutathione levels. CONCLUSION: The overall results demonstrated that the extracts exhibited significant antioxidant and protective effects in liver and brain liver homogenates.

Afrostyrax lepidophyllus extracts exhibit in vitro free radical scavenging, antioxidant potential and protective properties against liver enzymes ion mediated oxidative damage.

BACKGROUND: Several studies described the phytochemical constituents of plants in relation with the free radical scavenging property and inhibition of lipid peroxidation. This study investigated the in vitro antioxidant property, and the protective effects of ethanolic and aqueous ethanol extract of the leaves and barks of Afrostyrax lepidophyllus (Huaceae) against ion mediated oxidative damages. METHODS: Four extracts (ethanol and aqueous-ethanol) from the leaves and barks of A. lepidophyllus were used in this study. The total phenols content, the antiradical and antioxidant properties were determined using standard colorimetric methods. RESULTS: The plant extracts had a significant scavenging potential on the 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), nitrite oxide (NO) and 2,2-azinobis (3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radicals with the IC50 varied between 47 and 200 µg/mL depending on the part of plant and the type of extract. The ethanol extract of A. lepidophyllus bark (GEE) showed the highest polyphenolic (35.33 ± 0.29) and flavonoid (12.00 ± 0.14) content. All the tested extracts demonstrated a high protective potential with the increased of superoxide dismutase, catalase and peroxidase activities. CONCLUSION: Afrostyrax lepidophyllus extracts exhibited higher antioxidant potential and significant protective potential on liver enzymes.

In vitro antioxidant and anti-lipoperoxidative activities of bark extracts of Xylopia aethiopica against ion-mediated toxicity on liver homogenates.

Reactive oxygen species (ROS), products of normal cell metabolism may cause damage to biological macromolecules leading to severe health threats when they are present in high concentrations. Aromatic plants contain phytochemicals rich of antioxidants that prevent oxidant formation or scavenge oxidants produced under oxidative stress conditions. In the present study, we investigated the free radical scavenging effects, the antioxidant and ion toxicity preventive effect of Xylopia aethiopica (X. aethiopica), a plant of the family of Annonaceae used as spice in Cameroon. The scavenging properties of extracts of X. aethiopica were tested on 2, 2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), hydroxyl (OH), 2,2'-azinobis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radicals. The total antioxidant capacity was assayed by ferric reducing ability of plasma (FRAP), phosphomolybdenum antioxidant power (PAP), reduction assays. The protective potential was carried on superoxide dismutase (SOD), catalase and peroxidases. The results showed that both the ethanolic (BEE) and the hydroethanolic (BEH) extracts from the barks of X. aethiopica scavenged all the tested radicals. The sample BEH showed the highest total antioxidant capacity both in the FRAP and the PAP. This result was positively correlated to its higher phenolic content (30.74±0.44 CAE/g dried extract). The higher protective capacity of BEH on SOD, catalase and peroxidase activities was comparable to that of the vitamin C used as standard. In conclusion, X. aethiopica has a higher antioxidant and protective potential against ion-mediated oxidative damage and may be considered as a potential drug against metal-mediated toxicity.

In vitro antioxidant properties, free radicals scavenging activities of extracts and polyphenol composition of a non-timber forest product used as spice: Monodora myristica.

BACKGROUND: Excessive production of free radicals causes direct damage to biological molecules such as DNA, proteins, lipids, carbohydrates leading to tumor development and progression. Natural antioxidant molecules from phytochemicals of plant origin may directly inhibit either their production or limit their propagation or destroy them to protect the system. In the present study, Monodora myristica a non-timber forest product consumed in Cameroon as spice was screened for its free radical scavenging properties, antioxidant and enzymes protective activities. Its phenolic compound profile was also realized by HPLC. RESULTS: This study demonstrated that M. myristica has scavenging properties against DPPH(•), OH(•), NO(•), and ABTS(•) radicals which vary in a dose depending manner. It also showed an antioxidant potential that was comparable with that of Butylated Hydroxytoluene (BHT) and vitamin C used as standard. The aqueous ethanol extract of M. myristica barks (AEH); showed a significantly higher content in polyphenolic compounds (21.44 ± 0.24 mg caffeic acid/g dried extract) and flavonoid (5.69 ± 0.07 quercetin equivalent mg/g of dried weight) as compared to the other studied extracts. The HPLC analysis of the barks and leaves revealed the presence of several polyphenols. The acids (3,4-OH-benzoic, caffeic, gallic, O- and P- coumaric, syringic, vanillic), alcohols (tyrosol and OH-tyrosol), theobromine, quercetin, rutin, catechine and apigenin were the identified and quantified polyphenols. All the tested extracts demonstrated a high protective potential on the superoxide dismutase (SOD), catalase and peroxidase activities. CONCLUSION: Finally, the different extracts from M. myristica and specifically the aqueous ethanol extract reveal several properties such as higher free radical scavenging properties, significant antioxidant capacities and protective potential effects on liver enzymes.

Free radicals quenching potential, protective properties against oxidative mediated ion toxicity and HPLC phenolic profile of a Cameroonian spice: Piper guineensis.

Considerations on antioxidants derived from plants have continuously increased during this decade because of their beneficial effects on human health. In the present study we investigated the free radical scavenging properties of extracts from Piper guineense (P. guineense) and their inhibitory potentials against oxidative mediated ion toxicity. The free radical quenching properties of the extracts against [1,1-diphenyl-2-picrylhydrazyl (DPPH•), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS•), hydroxyl radical (HO•), nitric oxide (NO•)] radical and their antioxidant potentials by FRAP and phosphomolybdenum were determined as well as their protective properties on liver enzymes. The phenolic profile was also investigated by HPLC. The results obtained, revealed that the extracts significantly inhibited the DPPH, NO, HO and ABTS radicals in a concentration depending manner. They also showed a significant ferrous ion chelating ability through FRAP and phosphomolybdenum antioxidant potential. Their polyphenol contents varied depending on the type of extracts and the solvent used. The hydroethanolic extracts (FFH) and the ethanolic extracts (FFE) of P. guineense leaves showed the higher level of phenolic compounds respectively of 21.62 ± 0.06 mg caffeic acid/g dried extract (CAE/g DE) and 19.01 ± 0.03 CAE/g DE. The HPLC phenolic compounds profile revealed a higher quantity of Eugenol, quercetin, rutin and catechin in the stem than in the leaves. The presence of these molecules could be responsible of the protective potentials of P. guineense extracts against lipid peroxidation and SOD, catalase and peroxidase. In conclusion, P. guineense extracts demonstrated significant antioxidant property and may be used as a prospective protector against metal related toxicity.

Determinants of fructosamine levels in a multi-ethnic Sub-Saharan African population.

BACKGROUND AND PURPOSE: Fructosamine provides an estimate of diabetes control over a shorter period than HbA1c, and has been proposed as a suitable parameter to monitor glycemic control in low-income countries. The aim of this study was to investigate determinants of fructosamine levels in an urban non-diabetic population of Cameroon. METHODS: This was a cross-sectional study including 437 healthy adults with no known history of diabetes mellitus, aged 40 years and above, recruited from the ten administrative regions, representing major ethnic groups in the country. Plasma glucose and fructosamine were measured after an overnight fasting. Univariable and multivariable analyses were used to investigate the factors associated with fructosamine measurements. RESULTS: Fructosamine levels ranged from 68.2 to 940.8 µmol/l with a mean (standard deviation) of 294.4 (131.3) µmol/l. These levels varied significantly across regions and were higher in men than in women (p=0.001) and in those with screen-detected diabetes than in those with normoglycemia (p<0.0001). There was a negative correlation between fructosamine and body mass index (r=-0.15, p=0.009), and a positive correlation with fasting plasma glucose (FPG) (r=0.37, p<0.0001) and total bilirubinemia (r=0.21, p<0.0001). In multivariable model, sex, BMI, FPG, total bilirubine and screen-detected diabetes were no longer associated with fructosamine levels. CONCLUSION: Fructosamine was not independently associated with age, sex, ethnicity, and the glycemic status. Further studies need to be carried out to better elucidate all the factors determining the measurement of fructosamine in order to accurately interpret its values in diabetic populations.

In vitro antioxidant properties, free radicals scavenging activities of extracts and polyphenol composition of a non-timber forest product used as spice: Monodora myristica

BACKGROUND: Excessive production of free radicals causes direct damage to biological molecules such as DNA, proteins, lipids, carbohydrates leading to tumor development and progression. Natural antioxidant molecules from phytochemicals of plant origin may directly inhibit either their production or limit their propagation or destroy them to protect the system. In the present study, Monodora myristica a non-timber forest product consumed in Cameroon as spice was screened for its free radical scavenging properties, antioxidant and enzymes protective activities. Its phenolic compound profile was also realized by HPLC. RESULTS: This study demonstrated that M. myristica has scavenging properties against DPPH',OH',NO', and ABTS'radicals which vary in a dose depending manner. It also showed an antioxidant potential that was comparable with that of Butylated Hydroxytoluene (BHT) and vitamin C used as standard. The aqueous ethanol extract of M. myristica barks (AEH); showed a significantly higher content in polyphenolic compounds (21.44 ± 0.24 mg caffeic acid/g dried extract) and flavonoid (5.69 ± 0.07 quercetin equivalent mg/g of dried weight) as compared to the other studied extracts. The HPLC analysis of the barks and leaves revealed the presence of several polyphenols. The acids (3,4-OH-benzoic, caffeic, gallic, O- and P- coumaric, syringic, vanillic), alcohols (tyrosol and OH-tyrosol), theobromine, quercetin, rutin, catechine and apigenin were the identified and quantified polyphenols. All the tested extracts demonstrated a high protective potential on the superoxide dismutase (SOD), catalase and peroxidase activities. CONCLUSION: Finally, the different extracts from M. myristica and specifically the aqueous ethanol extract reveal several properties such as higher free radical scavenging properties, significant antioxidant capacities and protective potential effects on liver enzymes.

Antiproliferative activity and induction of apoptosis by Annona muricata (Annonaceae) extract on human cancer cells.

BACKGROUND: Annona muricata (A. muricata) is widely distributed in Asia, Africa and South America. Different parts of this plant are used to treat several diseases in Cameroon. The aim of this study is to determine the in vitro anti-proliferative effects and apoptotic events of A. muricata extracts on HL-60 cells as well as to quantify its phenols content. METHODS: The cell viability was measured by using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay while the changes in morphology of HL-60 cells, membrane mitochondrial potential (MMP) and the cell cycle were used for assessment apoptosis induction. RESULTS: The results show that the concentration of phenols, flavonoids and flavonols in the extracts varied depending on the part of the plant. All the extracts tested inhibited the proliferation of HL-60 cells in a concentration dependent manner with IC50 varied from 6-49 µg/mL. The growth inhibition of the cells by extracts was associated with the disruption of MMP, reactive oxygen species (ROS) generation and the G0/G1 cell arrest. CONCLUSION: These findings suggest that the extracts from A. muricata have strong antiproliferation potential and can induce apoptosis through loss of MMP and G0/G1 phase cell arrest.

Fruits and barks extracts of Zanthozyllum heitzii a spice from Cameroon induce mitochondrial dependent apoptosis and Go/G1 phase arrest in human leukemia HL-60 cells.

BACKGROUND: Zanthoxylum heitzii is a spice used to prepare several dishes and to treat tumors, syphilis, malaria, cardiac palpitations, urogenital infections in the west region of Cameroon, but the antitumor mechanisms and chemical composition are not yet investigated. This study was aimed to determine the antiproliferative effects of four extracts from the fruits and barks of Zanthoxyllum heitzii (Rutaceae) on apoptosis in human promyelocytic cells, their mechanisms and the chemical composition. The 3-(4, 5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the fifty percent inhibition (IC50) concentration of the cell lines after treatment. The effect on morphology was observed using a light or fluorescence microscopy. The rate of apoptosis and the cell cycle were measured using flow cytometry (FCM). The phytochemical analysis of the extract was carried with HPLC/MS methods. RESULTS: The phytochemical analysis of the extracts indicated the presence of four known polyphenols (Syringic acid, Juglon, Luteolin and Myricetin) in both fruits and barks of Z. heitzii but in different quantities. Syringic acid and Myricetin concentrations were between 17-21 fold higher in the fruits than the stem bark. Rhamnetin (393.35 µg/mL) and Oleuropein (63.10 µg/mL) were identified only in the stem barks of Z. heitzii. Among the four extracts tested for cytotoxicity properties, only the methanol extract of fruits and barks significantly inhibited cell proliferation of HL-60 cells with IC50 value of 20 µg/mL and 12 µg/mL respectively. HL-60 cells treated with Z. heitzii extracts significantly produced reactive oxygen species (ROS) with concurrent loss of mitochondrial membrane potential (MMP). Modifications in the DNA distribution and enhanced of G1/G0 phase cell cycle arrest were observed in a concentration dependent manner. CONCLUSIONS: Polyphenols from Z. heitzii plant exert inhibitory effect on HL-60 cells through the reactive oxygen species (ROS) generation, loss of mitochondrial membrane potential and cell cycle destabilization.

Phenolic Content of Hypodaphnis Zenkeri and Its Antioxidant Effects against Fenton Reactions' Mediated Oxidative Injuries on Liver Homogenate.

Under oxidative stress conditions, endogenous antioxidant defenses are unable to completely inactivate the free radicals generated by an excessive production of reactive oxygen species (ROS). This state causes serious cell damage leading to a variety of human diseases. Natural antioxidants can protect cells against oxidative stress. Hypaodaphnis zenkeri (H. zenkiri) is a plant consumed as a spice in the Cameroonian diet, and its bark has been used in traditional medicine for the treatment of several diseases. The present study aims at investigating the antioxidant activity, which includes free radical scavenging and protective properties of an extract from H. Zenkiri against oxidative damage on a liver homogenate. The free radical assays determined the scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), nitrite oxide (NO) and 2,2-azinobis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radicals and the enzymes, whose protection was to be considered in the liver homogenate, including superoxide dismutase, catalase, and peroxidase. The antioxidative activities were studied using the ferric reducing antioxidant power (FRAP), reductive activity, and phosphomolybdenum antioxidant power (PAP) methods. In addition, the phenolic contents of the extracts were examined. The results showed that these extracts demonstrated significant scavenging properties and antioxidant activities, with the hydro-ethanolic extract of the bark of H. zenkeri (EEH) being the most potent. This extract had the highest total polyphenol (21.77 ± 0.05 mg caffeic acid (CAE)/g dried extract (DE)) and flavonoids (3.34 ± 0.13 mg quercetin (QE)/g dried extract) content. The same extract had significantly greater protective effects on enzyme activities compared to other extracts. The high performance liquied chromatography (HPLC) profile showed higher levels of caffeic acid, OH-tyrosol acid, and rutin in the leaves compared to the bark of H. zenkeri. In conclusion, the ethanolic and hydro-ethanolic extracts of the bark and leaves from H. zenkeri showed an antioxidant and protective potential against oxidative damage.

Fruits and barks extracts of Zanthozyllum heitzii a spice from Cameroon induce mitochondrial dependent apoptosis and Go/G1 phase arrest in human leukemia HL-60 cells

BACKGROUND: Zanthoxylum heitzii is a spice used to prepare several dishes and to treat tumors, syphilis, malaria, cardiac palpitations, urogenital infections in the west region of Cameroon, but the antitumor mechanisms and chemical composition are not yet investigated. This study was aimed to determine the antiproliferative effects of four extracts from the fruits and barks of Zanthoxyllum heitzii (Rutaceae) on apoptosis in human promyelocytic cells, their mechanisms and the chemical composition. The 3-(4, 5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the fifty percent inhibition (IC50) concentration of the cell lines after treatment. The effect on morphology was observed using a light or fluorescence microscopy. The rate of apoptosis and the cell cycle were measured using flow cytometry (FCM). The phytochemical analysis of the extract was carried with HPLC/MS methods. RESULTS: The phytochemical analysis of the extracts indicated the presence of four known polyphenols (Syringic acid, Juglon, Luteolin and Myricetin) in both fruits and barks of Z. heitzii but in different quantities. Syringic acid and Myricetin concentrations were between 17-21 fold higher in the fruits than the stem bark. Rhamnetin (393.35 µg/mL) and Oleuropein (63.10 µg/mL) were identified only in the stem barks of Z. heitzii. Among the four extracts tested for cytotoxicity properties, only the methanol extract of fruits and barks significantly inhibited cell proliferation of HL-60 cells with IC50 value of 20 µg/mL and 12 µg/mL respectively. HL-60 cells treated with Z. heitzii extracts significantly produced reactive oxygen species (ROS) with concurrent loss of mitochondrial membrane potential (MMP). Modifications in the DNA distribution and enhanced of G1/G0 phase cell cycle arrest were observed in a concentration dependent manner. CONCLUSIONS: Polyphenols from Z. heitzii plant exert inhibitory effect on HL-60 cells through the reactive oxygen species (ROS) generation, loss of mitochondrial membrane potential and cell cycle destabilization.

Induction of mitochondrial dependent apoptosis and cell cycle arrest in human promyelocytic leukemia HL-60 cells by an extract from Dorstenia psilurus: a spice from Cameroon.

BACKGROUND: The use of edible plants is an integral part of dietary behavior in the West region of Cameroon. Dorstenia psilurus (Moraceae) is widely used as spice and as medicinal plant for the treatment of several diseases in Cameroon. The aim of this study is to investigate the cytotoxic and apoptotic potential of methanol extract of D. psilurus in human promyelocytic leukemia (HL-60) cells and prostate cancer (PC-3) cells. METHODS: Cytotoxicity of D. psilurus extract was tested in HL-60 and PC-3 cells using 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay and flow cytometric methods RESULTS: The methanol extract of D. psilurus have significant in vitro cytotoxic activity in HL-60 cells and PC-3 cells with IC50 value of 12 ± 1.54 µg/ml and 18 ± 0.45 µg/ml respectively after 48 h. The mechanism of antiproliferative activity showed that after 24 h, D. psilurus extract induces apoptosis on HL-60 cells by the generation of reactive oxygen species (ROS) along with concurrent loss of mitochondrial membrane potential, modification in the DNA distribution and enhance of G2/M phase cell cycle. CONCLUSION: The extract induces apoptosis of HL-60 cells associated with ROS production, loss of mitochondrial membrane potential and apoptotic DNA fragmentation.