Antioxidant Activities of Some Cameroonian Plants Extracts Used in the Treatment of Intestinal and Infectious Diseases

Antioxidant activity test using two different methods namely 2,2-diphenyl-1-picrylhydrazyl and 2,2''-azinobis(3-ethylbenzothialozinesulfonate) diammonium salt free radical scavenging test has been carried out on three Cameroonian plant extracts used in the treatment of intestinal and infectious diseases: Pittosporum mannii Hook f. (Pittosporaceae), Vepris heterophylla R. Letouzey (Rutaceae) and Ricinodendron heudelotii (Baill) Pierre ex Pax (Euphorbiaceae). Results of this study in the 2,2-diphenyl-1-picrylhydrazyl scavenging test show that the ethyl acetate extract of P. mannii and the methanol extract of V. heterophylla exhibit high free radical scavenging activities with IC₅₀ values of 177.74 and 204.69 μg/ml, respectively while the methanol/dichloromethane (1+1) extract of R. heudelotii showed weak free radical scavenging activities as compared to Trolox (939.19 μg/ml) used as standard. In the same manner, 2,2''-azinobis(3-ethylbenzothialozinesulfonate) diammonium salt radical scavenging test of these extracts was in accordance of the result of 2,2-diphenyl-1-picrylhydrazyl test. The antioxidant properties of these extracts probably explain partly, the use of these plants in traditional medicine for the treatment of infectious diseases and inflammations.     

Phytochemical Analysis and In vitro Antioxidant Studies of Plumeria obtusa L. Leaves

Plumeria (Apocynaceae) is a genus comprising mostly of lactiferous trees and deciduous shrubs. Plumeria obtusa L. is a widely available but pharmacologically lesser explored species of this genus. Thus, present research was undertaken to determine the phytochemical constituents and the antioxidant potential of the methanol extract and fractions of the plant. The antioxidant potential was determined using 1,1-diphenyl-2-picrylhydrazyl radical scavenging and inhibition of lipid peroxidation assay.     

Investigation of Phytochemical and Antioxidant Properties of Methanol Extract and Fractions of Ballota limbata (Lamiaceae)

Ballota limbata (Lamiaceae) has been used for its antispasmodic, antiulcer, diuretic, vermifuge and sedative effects in folk medicine. However, little is known about how does it work to produce these therapeutic actions. Present research investigated phytochemical components and antioxidant properties of methanol extract and different fractions of Ballota limbata. In this study, phytochemical investigation was done by performing different chemical tests. Here, antioxidant property of the extract and fractions was investigated by using 1,1-diphenyl-2-picryl hydrazyl radical scavenging activity, total antioxidant activity by the phosphomolybdenum method, linoleic acid peroxidation, ferric thiocyanate analysis and ferric-reducing antioxidant power. Methanol extract and fractions showed presence of numerous chemical principles including alkaloids, cardiac glycosides, tannins and flavonoids. The ethyl acetate fraction exhibited higher scavenging activity compared to the other fractions under investigation. This fraction displayed 84.16±1.02% 1,1-diphenyl-2-picryl hydrazyl radical inhibition at a dose of 60 μg/ml. IC₅₀ for 1,1-diphenyl-2-picryl hydrazylradical-scavenging activity was 13.53±0.22 μg/ml, relative to the standard, butylatedhydroxytoluene, having IC₅₀ of 12.33±0.88 μg/ml. Thus, Ballota limbata showed significant antioxidant activity, which may contribute in the mechanism of above pharmacological actions.     

Hepatoprotective and Antioxidant Activities of Desmodium Triquetrum DC

The hepatoprotective and antioxidant activities of ethanol extract of Desmodium triquetrum DC leaf were investigated against carbon tetrachloride (1 ml/kg i.p) induced hepatic damage in rats at a dose of 200 mg/kg body weight p.o. The test extract significantly (P<0.05) reduced the elevated levels of serum transaminases, alkaline phosphatase, bilirubin and reversed the antioxidant enzyme and non-enzyme levels. It dose dependently inhibited thiobarbuturic acid induced lipid peroxidation in vitro (IC(50)=59.9 μg/ml). Histopathological studies provided supportive evidence for biochemical analysis. Silymarin (25 mg/kg) is a known hepatoprotective drug used as a reference drug. The results indicated that D. triquetrum has potent hepatoprotective and antioxidant activity that may be due to the presence of flavonoids in the plant.     

Anti-Inflammatory,Antioxidant, Anti-Angiogenic and Skin Whitening Activities of Phryma leptostachya var. asiatica Hara Extract

This work aimed to assess some pharmacological activities of P. leptostachya var. asiatica Hara. The dried roots of P. leptostachya var. asiatica Hara were extracted with 70% ethanol to generate the powdered extract, named PLE. Anti-angiogenic activity was detected using chick chorioallantoic membrane (CAM) assay. In vitro anti-inflammatory activity was evaluated via analyzing nitric oxide (NO) content, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Antioxidant activity was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and reactive oxygen species (ROS) level in the stimulated macrophage cells. Matrix metalloproteinase-9 (MMP-9) and -2 (MMP-2) activities in the culture media were detected using zymography. PLE exhibits an anti-angiogenic activity in the CAM assay, and displays an inhibitory action on the generation of NO in the LPS-stimulated macrophage cells. In the stimulated macrophage cells, it is able to diminish the enhanced ROS level. It can potently scavenge the stable DPPH free radical. It suppresses the induction of iNOS and COX-2 and the enhanced MMP-9 activity in the stimulated macrophage cells. Both monooxygenase and oxidase activities of tyrosinase were strongly inhibited by PLE. Taken together, the dried roots of P. leptostachya var. asiatica Hara possess anti-angiogenic, anti-inflammatory, antioxidant and skin whitening activities, which might partly provide its therapeutic efficacy in traditional medicine.     

In vitro Antioxidant and Antibacterial Activities of Methanol Extract of Kyllinga nemoralis

The present study was designed to evaluate the antioxidant and antibacterial activity of methanol extract of Kyllinga nemoralis. Six different in vitro antioxidant assays including 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide anion radical, hydrogen peroxide radical, ferric reducing antioxidant power assay and reducing power were carried out to ensure the scavenging effect of the plant on free radicals. In addition, total antioxidant capacity assay, total phenolic contents, tannins, flavonoids and flavonol contents of the plant were also analysed by the standard protocols. Kyllinga nemoralis exhibited high antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl assay (IC₅₀= 90 μg/ml), superoxide radical scavenging assay (IC₅₀= 180 μg/ml) and hydrogen peroxide radical scavenging assay (IC₅₀= 200 μg/ml), compared with standards. These observations provide comprehensible supporting evidence for the antioxidant potential of the plant extract. Reducing power (IC₅₀= 213.16 μg/ml) and hydroxyl radical scavenging activity (IC₅₀= 223 μg/ml) of the plant extract was remarkable. The methanol extract of K. nemoralis exhibited significant antimicrobial activity against Gram-positive human pathogenic bacteria. Standard in vitro antioxidant assays assessed the electron donating ability of the plant extract in scavenging free radicals. The inhibitory effect of the plant extract against bacterial pathogens may be due to the presence of phytochemicals. Thus, the results suggest that Kyllinga nemoralis is a potential source of antioxidants and could serve as the base for drug development.     

In vitro Antioxidant Studies of Fruits of Artemisia nilagirica (Clarke) Pamp

Antioxidant potential of fruits of Artemisia nilagirica was studied using different in vitro models like 1,1-diphenyl-2-picryl hydrazyl, 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate), nitric oxide, superoxide, hydroxyl radical and lipid peroxidation. Both the ethanol and aqueous extracts of A. nilagirica fruits at 500 μg/ml showed maximum scavenging activity (89.33% and 89.14%) in quenching 1,1-diphenyl-2-picryl hydrazyl radical. The ethanol extract showed better scavenging activity (69.78%) of 1,1-diphenyl-2-picryl hydrazyl radical followed by the scavenging of nitric oxide radical (73.25%) compared to aqueous extract. In contrast, hydroxyl and superoxide radicals were effectively scavenged by aqueous extract. Total antioxidant capacity of ethanol and aqueous extracts at 500 μg/ml concentration was found to be 56.21 and 62.78 mg ascorbic acid equivalents, respectively. However, both the extracts showed only moderate lipid peroxidation inhibition activity. They were also found to contain considerable total phenols and flavonoids suggesting their role as an effective free radical scavenger. These findings suggest that phenolics and flavonoids in the fruits provide substantial antioxidant activity.     

Antimicrobial Activity and Phytochemical Constituents of Leaf Extracts of Cassia auriculata

Plants produce a wide variety of phytochemical constituents, which are secondary metabolites and are used either directly or indirectly in the pharmaceutical industry. ‘For centuries, man has effectively used various components of plants or their extracts for the treatment of many diseases, including bacterial infections. In the present study methanol, chloroform and aqueous extracts of Cassia auriculata leaf were subjected for antimicrobial activity by well-diffusion method against six bacterial strains namely Bacillus cereus, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Proteus mirabilis. The results revealed that the methanol and chloroform extracts exhibited strong inhibitory activity against all the tested organisms (zone of inhibition of 12-20 mm), except Pseudomonas aeruginosa (zone of inhibition 10 mm or nil). The aqueous extracts showed moderate activity by ‘Zone of inhibition ≤12 or nil). The extracts were screened for their phytochemical constituents by standard protocols’ and were shown to contain carbohydrates, proteins, alkaloids, flavonoids, steroids, saponins and tannins. The antibacterial activity of these extracts is possibly linked to the presence of flavonoids, steroid, saponins and/or tannins. Further studies are needed to determine the precise active principles from Cassia auriculata.     

Phytochemical Screening,Proximate Analysis and Antioxidant Activity of Dracaena reflexa Lam. Leaves

In the present study, the antioxidant activity of successive leaf extracts of Dracaena reflexa was investigated using the scavenging activity on 1,1-diphenyl-2-picrylhydrazyl and reducing power by ferric reducing antioxidant power assay. Methanol extract was found potent in both the assays. IC50 values of 1,1-diphenyl-2-picrylhydrazyl assay for methanol extract was 0.97 mg/ml and ferric reducing antioxidant power value for the same is 1.19. Phytochemical screening, proximate analysis and total phenolic content were also determined. Qualitative screening for phytochemical showed the presence of alkaloids, flavonoids, terpenoids, glycosides and saponins. Highest phenolic content was shown by methanol extract (49.69 mg gallic acid equivalent/g dry weight). Proximate analysis showed moisture content (3.31%), ash content (8.02%), crude fibre (1.31%), crude fat (0.97%), total protein (3.70%), total carbohydrate (86.01) and nutritive value (367.56 kcal/100 g), which would make it a potential nutraceutical. This study suggested that Dracaena reflexa, a potential natural free radical scavenger, which could find use as an antioxidative.     

Antioxidant Activity in the Extracts of Two Edible Aroids

Two neglected species of Araceae, Alocasia macrorhiza (Linn.) G. Don and Alocasia fornicata (Roxb.) Schott are important as food and ethno medicine in Asia and Africa. Their bioefficacy is documented in the Ayurveda. The solvent extracts of different edible parts of these two species like rhizomes, leaves, roots and stolons were screened for in vitro antioxidant properties using standard procedures. The successive extracts in hexane, benzene, toluene, chloroform, diethyl ether, ethyl acetate and water fraction exhibited IC₅₀ values in the following order, roots>rhizome>leaves for Alocasia macrorhiza and leaves>stolon for Alocasia fornicate, respectively in 2,2-diphenyl-1-picryl hydrazyl antioxidant inhibition assay. Maximum antioxidant activity was observed in diethyl ether extracts for both species. The IC₅₀ values were comparable with those of quercetine and ascorbic acid as standards. These results suggest that the two aroid species have antioxidant activity in their edible parts and should be extracted using diethyl ether solvent.     

Antioxidant Activity and Gas Chromatographic-Mass Spectrometric Analysis of Extracts of the Marine Algae,Caulerpa peltata and Padina Gymnospora

The results of our previous investigations on extracts of selected marine algae showed that Caulerpa peltata and Padina gymnospora had more promising antiproliferative and antioxidant activities than Gelidiella acerosa and Sargassum wightii. Based on these results, the more active chloroform extract of C. peltata and ethyl acetate extract of P. gymnospora were further analyzed for their constituents by using gas chromatography in tandem with mass spectrometry. The GC-MS analysis (GC % peak area given in parentheses) showed that fucosterol (12.45%) and L-(+)-ascorbic acid 2, 6-dihexadecanoate (8.13%) were the major compounds present in P. gymnospora ethyl acetate extract. On the other hand, C. peltata chloroform extract had 1-heptacosanol (10.52%), hexacosanol acetate (9.28%), tetradecyl ester of chloroacetic acid (7.22%), Z,Z-6, 28-heptatriactontadien-2-one (6.77%) and 10, 13-dimethyl-methyl ester of tetradecanoic acid (5.34%) as major compounds. Also described in the report are the beta-carotene bleaching inhibitory and total reducing activities of the chloroform and ethyl acetate extracts of C. peltata and P. gymnospora, respectively, relative to the other three extracts (aqueous, methanol, chloroform or ethyl acetate) of the two algae.     

Antifungal and Anticancer Activities of a Protein from the Mushroom Cordyceps militaris

The mushroom Cordyceps militaris has been used for a long time in eastern Asia as a nutraceutical and in traditional Chinese medicine as a treatment for cancer patients. In the present study, a cytotoxic antifungal protease was purified from the dried fruiting bodies of C. militaris using anion-exchange chromatography on a DEAE-Sepharose column. Electrophoretic analyses indicated that this protein, designated C. militaris protein (CMP), has a molecular mass of 12 kDa and a pI of 5.1. The optimum conditions for protease activity were a temperature of 37℃ and pH of 7.0~9.0. The enzyme activity was specifically inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride. Amino acid composition of intact CMP and amino acid sequences of three major peptides from a tryptic digest of CMP were determined. CMP exerted strong antifungal effect against the growth of the fungus Fusarium oxysporum, and exhibited cytotoxicity against human breast and bladder cancer cells. These results indicate that C. militaris represents a source of a novel protein that might be applied in diverse biological and medicinal applications.     

Phytochemical and pharmacological study of Ficus palmata growing in Saudi Arabia

Phytochemical study of the aerial parts of Ficus palmata utilizing liquid–liquid fractionation and different chromatographic techniques resulted in the isolation of a new isomer of psoralenoside namely, trans-psoralenoside (5) in addition to, one triterpene: germanicol acetate (1), two furanocoumarins: psoralene (2), bergapten (3), one aromatic acid vanillic acid (4) and the flavone glycoside rutin (6). Structures of the isolated compounds were established through physical, 1D- and 2D-NMR and MS data. The total extract and fractions of the plant were examined in vivo for its possible effects as hepatoprotective, nephroprotective, antiulcer and anticoagulant activities in comparison with standard drugs. Hepatoprotective activity was assessed via serum biochemical parameters including aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyl transpeptidase (GGT), alkaline phosphatase (ALP) and total bilirubin. Tissue parameters such as non-protein sulfhydryl groups (NP-SH), malonaldehyde (MDA) and total protein (TP) were also measured. In addition to tissue parameters, nephroprotective effect was evaluated by measuring the serum levels of sodium, potassium, creatinine and urea. Histopathological study for both liver and kidney cells was also conducted. Antiulcer activity was explored by observing stomach lesions after treatment with ethanol. Whole blood clotting time (CT) was taken as a measure for the anticoagulant activity of the extract. Antioxidant activity of the total extract and fractions of the plant was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) method and ascorbic acid as standard.     

Antioxidant and anti-inflammatory effects of Marrubium alysson extracts in high cholesterol-fed rabbits

The antioxidant and anti-inflammatory effects of hexane (HEXA), chloroform (CHLORO), ethyl acetate (EA) and total alcoholic (T. ALCOH) extracts of Marrubium alysson in hypercholesterolemic-fed rabbits were evaluated. Hypercholesterolemia was induced in male rabbits by high cholesterol diet (HCD) (350 mg/kg) for 8 weeks. Hypercholesterolemic rabbits were allocated into groups, treated with simvastatin (SIM 5 mg/kg), different extracts of M. alysson at two doses of 250, 500 mg/kg. A normal control group and an HCD control one were used for comparison. Lipid profile, as well as oxidized low density lipoprotein-cholesterol (ox-LDL-C), myeloperoxidase activity (MPO) and superoxide anion production (O₂•⁻), C-reactive protein (CRP) and monocyte chemoattractant protein-1 (MCP-1) were also evaluated. In addition, histological examination of ascending aorta was performed. We found dyslipidemia associated with significant increases in ox-LDL-C 123.5 ± 9.8 nmol MDA/mg non-HDL, MPO activity 0.08 ± 0.05 U/100 mg tissue and O₂•⁻ production 3.5 ± 0.3 nmol cytochrome C reduced/min/g tissue × 10⁻⁴ in hypercholerterolemic rabbits. In addition, there was a significant increase in CRP 6.6 ± 0.49 μmol/L and MCP-1 190.9 ± 6.4 pg/ml and its mRNA expression in HCD. Intima appeared thick with thick plaques surrounding the intima and luminal narrowing. SIM, EA and HEXA extracts of M. alysson had lipid lowering effect, decrease in ox-LDL-C, MPO, O₂•⁻, CRP and MCP-1 mRNA expression with improvement of the pathological picture. M. alysson enhanced the stability of plaque, had lipid lowering, anti-inflammatory and antioxidant activities.     

Antioxidant Studies on Ethanol Extracts from Two Selected Genera of Indian Lamiaceae

The present work is targeted to evaluate antioxidant activity of ethanol extracts from the leaves of Plectranthus mollis and Salvia officinalis belonging to family Lamiaceae using nitric oxide scavenging, hydrogen peroxide scavenging, ferric reducing antioxidant power assay and lipid peroxidation methods. The results of the study indicate that the leaf extracts of both the plants possess in vitro antioxidant activity. The higher amount of flavanoids and phenolic compounds may correspond to their greater antioxidant activity.     

Estimation of Total Phenols and Flavonoids in Extracts of Actaea spicata Roots and Antioxidant Activity Studies

Actaea spicata Linn. (Ranunculaceae) has been traditionally used for the treatment of various ailments such as rheumatism, inflammation, nerve diseases, lumbago, scrofula and chorea, but no systematic phytochemical and pharmacological work has ever been carried out on this potential plant. Preliminary phytochemical screening showed presence of phenols and flavonoids in A. spicata. Thus, the present investigation was undertaken to estimate total phenols and flavonoids in methanol extract of A. spicata roots, and its ethyl acetate fraction. In vitro antioxidant activity was also evaluated in the methanol extract and ethyl acetate fraction using DPPH method. Ethyl acetate fraction was found to contain twice the content of flavonoids and phenols in comparison to methanolic extract, whereas phenolic content in methanol extract was approximately similar to ethyl acetate fraction. A significant antioxidant activity, i.e., mean percentage inhibition of DPPH radical was observed in methanol extract and ethyl acetate fraction at the concentration of 10 μg/ml and 5 μg/ml respectively. Finally, it was suggested that polyphenols are responsible for antioxidant activity of A. spicata.     

Antioxidant and hepatoprotective activity of Cordia macleodii leaves

This investigation was undertaken to evaluate ethanolic extract of Cordia macleodii leaves for possible antioxidant and hepatoprotective potential. Antioxidant activity of the extracts was evaluated by four established, in vitro methods viz. 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging method, nitric oxide (NO) radical scavenging method, iron chelation method and reducing power method. The extract demonstrated a significant dose dependent antioxidant activity comparable with ascorbic acid. The extract was also evaluated for hepatoprotective activity by carbon tetrachloride (CCl₄) induced liver damage model in rats. CCl₄ produced a significant increase in levels of serum glutamate pyruvate transaminase (GPT), serum glutamate oxaloacetate transaminase (GOT), Alkaline Phosphatase (ALP) and total bilirubin. Pretreatment of the rats with ethanolic extract of C. macleodii (100, 200 and 400 mg/kg po) inhibited the increase in levels of GPT, GOT, ALP and total bilirubin and the inhibition was comparable with Silymarin (100 mg/kg po). The present study revealed that C. macleodii leaves have significant radical scavenging and hepatoprotective activities.     

Evaluation of Antioxidant Activity of Picrorhiza kurroa (Leaves) Extracts

Picrorhiza kurroa is a well-known herb in Ayurvedic medicine. Although it shows antioxidant, antiinflammatory and immunomodulatory activities, it is most valued for its hepatoprotective effect. The rhizomes are widely used against indigestion problems since ancient times due to improper digestive secretions. Aim of this study was to explore antioxidant study of P. kurroa leaves for a new source of naturally occurring antioxidants. Two pure compounds, luteolin-5-O-glucopyranoside (1) and picein (2) were isolated from butanol extract through column chromatography. Different extracts of P. kurroa leaves (ethanol, ethyl acetate, butanol) were quantified for isolated compound (2) by high-performance liquid chromatography. All the extracts and isolated compounds were evaluated for its antioxidant activity using two assays, 2,2-diphenyl-1-picrylhydrazyl radical and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) assay. The linear detection range was 1.56-200 μg/ml for picein. The limit of detection and limit of quantification for picein were 2.34 and 7.81 μg/ml, respectively. Butanol and ethyl acetate extract showed greater antioxidant activity as compare to ethanol extract. Compound 1 and ascorbic acid showed nearly similar antioxidant activity where as 2 showed no activity at standard concentration. The IC₅₀ values for 2,2-diphenyl-1-picrylhydrazyl radical and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) assay for ascorbic acid, compound 1, ethanol extract and its different fractions (ethyl acetate and butanol) were found to be 0.81, 1.04, 67.48, 39.58, 37.12 and 2.59, 4.02, 48.36, 33.24, 29.48 μg, respectively.     

In Vitro Anticancer Activity of the Root, Stem and Leaves of Withania Somnifera against Various Human Cancer Cell Lines

Withania Somnifera Dunal know as Ashwagandha belong Solanaceae family. It is extensively used in most of the Indian herbal pharmaceuticals and nutraceuticals. The current study, evaluate in vitro cytotoxicity in 50% ethanol extract of root, stem and leaves of Withania Somnifera against five human cancer cell lines of four different tissues i.e. PC-3, DU-145 (prostrate), HCT-15 (colon), A-549 (lung) and IMR-32 (neuroblastoma). Root, stem and leaves extracts showed cytotoxicity activity ranging 0-98% depending on the cell lines but maximum activity was found in 50% ethanol extract of leaves of Withania Somnifera. Ethanol extract of leaves obtained from treatments T2, T3, T4 and T5 showed strong activity against PC-3 and HCT-15 with 80-98% growth inhibition, while the 50% ethanol extract of leaves from T1 treatment showed a minimum of 39% and T3 treatment showed a maximum of 98% growth inhibition against HCT-15. This investigation is the first report of the anticancer activity in various parts of Withania Somnifera cultivated in fly ash amended soil.     

In vitro Antioxidant Potential in Sequential Extracts of Curcuma caesia Roxb. Rhizomes

Present study deals with antioxidant potential of sequential extracts of fresh and dried rhizomes of Curcuma caesia, using solvents viz., hexane, petroleum ether, benzene, chloroform, ethyl acetate, methanol and water, which was analyzed by 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, total antioxidant capacity, ferric reducing activity and thiobarbituric acid reactive species assay. Total phenol content was estimated by the Folin-Ciocalteau method. C. caesia showed significant antioxidant activity in chloroform, benzene and ethyl acetate extracts. The chloroform extract was highly effective as free radical scavengers, electron-donating agents and reducing molybdate ions except for reducing lipid peroxidation. The highest total phenol content was also exhibited by chloroform and benzene extracts. Antioxidant potential expressed by C. caesia in the sequential extracts could be effectively utilized for identification of the bioactive compounds for future phytopharmacological applications.