Identification and characterization of anti-diabetic principle in Senna alata (Linn.) flower using alloxan-induced diabetic male Wistar rats.

ETHNO-PHARMACOLOGICAL RELEVANCE: The age-long folkloric use of Senna alata flower (SAF) was recently substantiated with scientific evidence. However, the study did not account for the anti-diabetic principle(s) in SAF. AIM OF THE STUDY: The study aimed to identify and characterize the bioactive principle(s) responsible for the anti-diabetic activity in SAF. MATERIALS AND METHODS: Ninety-one male Wistar rats were used for the two phases of this study. In phase 1, forty-two of these were allotted into six groups (A-F) of seven rats each. Animals in group A received distilled water while those in groups B-F were made diabetic by treatment with 150 mg/kg body weight (b.w.) of alloxan. Group B received 0.5 mL of distilled water; C, D and E were treated each with 75 mg/kg b.w. of ethyl acetate, n-butanol and aqueous residual fractions of SAF, while F received 2.5 mg/kg b.w. of glibenclamide. In the second phase, forty-nine rats were assigned into seven groups (A-G) of seven rats each. Group A received distilled water. Animals in Groups B-G were also made diabetic by alloxan treatment. B received 0.5 mL of distilled water; C, D, E and F were treated with 5.77, 25.96, 15.40, 27.87 mg/kg b.w (equivalent dose of 75 mg/kg b.w.) of sub-fractions obtained from the ethyl acetate fraction of SAF respectively whereas G received 2.5 mg/kg b.w. of glibenclamide. Fasting blood glucose (FBG), serum lipids, albumin, globulin, liver glycogen, urine ketone, hexokinase and glucose-6-phosphate dehydrogenase activities, α-glucosidase and α-amylase inhibitory activities and cardiac function indices were evaluated using standard methods. Compounds D, E and F isolated from ethyl acetate sub-fraction B were evaluated for in vitro anti-diabetic activity. The structure of the anti-diabetic compound was identified using FTIR, 1H-NMR, 1³C-NMR, HCOSY, HSQC and HMBC. Data were subjected to Analysis of Variance and Duncan Multiple Range Test at p < 0.05. RESULTS: Alloxan treatment increased the levels of FBG, total cholesterol, LDL-cholesterol, VLDL-cholesterol, urine ketone and cardiac function indices and reduced the levels of globulin, albumin, HDL-cholesterol, globulin, liver glycogen, hexokinase and glucose-6-phosphate dehydrogenase activities. Ethyl acetate fraction and sub-fraction B reversed the level and/or activities of these biochemical indices to levels and/or activities that compared favourably with the distilled water treated non-diabetic animals. Of the three compounds (D, E and F) that were obtained from the sub-fraction B, compound E which was Emodin (1, 3, 8-trihydroxy-6-methylanthraquinone) produced the highest α-glucosidase and α-amylase inhibitory activities. CONCLUSION: Emodin is one of the bioactive constituents present in Senna alata flower.

Isolation of flavonoids from Musa acuminata Colla (Simili radjah, ABB) and the in vitro inhibitory effects of its leaf and fruit fractions on free radicals, acetylcholinesterase, 15-lipoxygenase, and carbohydrate hydrolyzing enzymes.

Musa species are used traditionally for the management of many diseases. The study evaluated and compared anticholinesterase, anti-inflammatory, antioxidant, and antidiabetic activities of Musa acuminata (Simili radjah, ABB) fruits and leaves fractions and characterized the bioactive compounds using HPTLC-HRMS and NMR. Leaf fractions gave the higher biological activities than the fruit. Ethyl acetate fraction of the leaf had the highest total phenolic content (911.9 ± 1.7 mg GAE/g) and highest 2,2-diphenyl-1-picrylhydrazyl (DPPH· ) scavenging activity (IC50, 9.0 ± 0.4 µg/ml). It also gave the most effective inhibition of acetylcholinesterase (IC50, 404.4 ± 8.0 µg/ml) and α-glucosidase (IC50, 4.9 ± 1.6 µg/ml), but a moderate α-amylase inhibition (IC50, 444.3 ± 4.0 µg/ml). The anti-inflammatory activity of n-butanol (IC50, 34.1 ± 2.6 µg/ml) and ethyl acetate fractions (IC50 , 43.1 ± 11.3 µg/ml) of the leaf were higher than the positive control, quercetin (IC50 , 54.8 ± 17.1 µg/ml). Kaempferol-3-O-rutinoside and quercetin-3-O-rutinoside (rutin) were identified as the bioactive compounds with antioxidant and antidiabetic activities from the ethyl acetate fraction of M. acuminata leaf. PRACTICAL APPLICATIONS: All parts of Musa acuminata are known to be useful ethnomedicinally even as food. The leaves are mostly used to serve food and used for wrapping purposes. However, this study concluded that M. acuminata leaf is rich in bioactive flavonoids such as kaempferol-3-O-rutinoside and rutin, with relatively high antioxidative, antidiabetic, and anti-inflammatory activities. Therefore, aside the fact that the leaves can serve as potential drug leads for pharmaceutical industries, it can also be embraced in the food sector to produce supplements and/or nutraceuticals in the management of Alzheimer's, diabetes and other inflammatory diseases.

Screening of Potential Phytocompounds From Euclea crispa (Thunb.) Leaves Targeting Human Epidermal Growth Factor Receptor 2 (HER2) Signaling Pathway.

BACKGROUND: Overexpression of human epidermal growth factor receptor 2 (HER2) plays an important role in the development and progression in a variety of cancers and it is a novel therapeutic target for breast cancer and ovarian cancer. Euclea crispa (E. crispa) is a South African medicinal plant in the family Ebenaceae used in the management of different human diseases and disorders. AIMS: The aim of this study was to evaluate the potential inhibitors against HER2 from hexane extract of E. crispa leaves. MATERIALS AND METHODS: Chemical fingerprinting method was used to identify the presence of natural compounds from the extract whereas their inhibitory activities were analyzed by molecular docking analysis against HER2. Absorption, distribution, metabolism, and excretion (ADME) properties also predicted to establish the pharmacokinetics and pharmacodynamics profiles of the selected compounds. RESULTS: The molecular docking analysis expressed that phenyl glucuronide, hydrocortisone acetate, and 6-(4,6-dioxo-1,4,5,6-tetrahydropyrimidin-2-yl-amino)hexanoic acid trifluoroacetate possess good inhibitory activities with good glide score of -6.63, -5.41, and -5.40 and glide energy of -35.03, -42.51, and -31.38 kcal/mol, respectively when compared with standard Food and Drug Administration-approved drug and other compounds. All the screened compounds were within the acceptable and permissible limits of ADME properties. CONCLUSION: Thus, from this study it can be concluded that, these screened natural compounds from E. crispa leaves may serve as potential inhibitors for HER2 and they might lead to development of new therapeutic agents against cancer and its associated complications.

A Review of Plants Used in South African Traditional Medicine for the Management and Treatment of Hypertension.

South Africa contains 9% of the world's higher plants, and despite its rich biodiversity, it has one of the highest prevalence of hypertension in Africa. This review provides information on medicinal plants embraced in South Africa for hypertension management, with the aim of reporting pharmacological information on the indigenous use of these plants as antihypertensives. This review not only focuses on the activity of antihypertensive medicinal plants but also reports some of its phytochemical constituents and other ethnopharmacological and therapeutic properties. Information obtained from scientific and or unpublished databases such as Science Direct, PubMed, SciFinder, JSTOR, Google Scholar, Web of Science, and various books revealed 117 documented antihypertensive plant species from 50 families. Interestingly, Asteraceae topped the list with 16 species, followed by Fabaceae with 8 species; however, only 25% of all plant species have demonstrated antihypertensive effects originating from both in vitro and in vivo studies, lending credence to their folkloric use. Only 11 plant species reportedly possess antihypertensive properties in animal models, with very few species subjected to analytical processes to reveal the identity of their bioactive antihypertensive compounds. In this review, we hope to encourage researchers and global research institutions (universities, agricultural research councils, and medical research councils), particularly those showing an interest in natural products, for the need for concerted efforts to undertake more studies aimed at revealing the untapped potential of these plants. These studies are very important for the development of new pharmaceuticals of natural origin useful for the management of hypertension.

Antioxidant and Inhibitory Effects of Saponin Extracts from Dianthus basuticus Burtt Davy on Key Enzymes Implicated in Type 2 Diabetes In vitro.

CONTEXT: Dianthus basuticus is a plant of South African origin with various acclaimed pharmaceutical potentials. AIMS: This study explored the antioxidant and antidiabetic activities of saponin extract from D. basuticus in vitro. MATERIALS AND METHODS: Antioxidant activity of saponin was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (*NO)-free radical scavenging activity while antidiabetic potentials were measured by the α-amylase and α-glucosidase inhibitory activities of the saponin extract. RESULTS: The results showed that the saponin extract, compared with quercetin, displayed better DPPH (IC50 = 6.95 mg/ml) and NO (IC50 = 3.31 mg/ml) radical scavenging capabilities. Similarly, the saponin extracts elicited stronger α-glucosidase (IC50 = 3.80 mg/ml) and moderate α-amylase (IC50 = 4.18 mg/ml) inhibitory activities as compared to acarbose. Saponin exhibited a competitive mode of inhibition on α-amylase with same maximum velocity (Vmax) of 0.0093 mM/min for saponin compared with control 0.0095 mM/min and different the Michaelis constant (Km) values of 2.6 × 10-6 mM and 2.1 × 10-5 mM, respectively, while for α-glucosidase, the inhibition was uncompetitive, Vmax of 0.027 mM/min compared with control 0.039 mM/min and Km values of 1.02 × 10-6 mM and 1.38 × 10-6 mM, respectively. The gas chromatography-mass spectrometric analysis revealed the presence of bioactive like ß- and α-amyrin, 3-O-methyl-D-glucose, methyl commate, and olean-12-en-3-beta-ol. CONCLUSION: Overall, the data suggested that the saponin extract from D. basuticus has potentials as natural antioxidants and antidiabetics. SUMMARY: Saponin extract from Dianthus basuticus displayed promising antidiabetic and antioxidant activitySaponin competitively and uncompetitively inhibited a-amylase and a-glucosidase, respectivelyThe stronger inhibition of α-glucosidase and moderate inhibition of α-amylase by saponin extract from D. basuticus is promising good antidiabetes compared with existing drugs with associated side effects. Abbreviations used: DPPH: 2,2-diphenyl-1-picrylhydrazyl, Km: The Michaelis constant, Vmax: Maximum velocity, ROS: Reactive oxygen species, NIDDM: Non-insulin-dependent diabetes mellitus, UFS: University of the Free State, GC-MS: Gas chromatography-mass spectrometric, MS: Mass spectrometry, NIST: National Institute of Standards and Technology, DNS: 3,5-dinitrosalicylic acid, NO: Nitric oxide, RNS: Reactive nitrogen species, PNPG: p-Nitrophenyl-α-D-glucopyranoside.

KINETICS OF MODULATORY ROLE OF Cyperus esculentus L. ON THE SPECIFIC ACTIVITY OF KEY CARBOHYDRATE METABOLIZING ENZYMES.

BACKGROUND: The continuous search for new lead compounds as viable inhibitors of specific enzymes linked to carbohydrate metabolism has intensified. Cyperus esculentus L. is one of the therapeutically implicated botanicals against several degenerative diseases including diabetes mellitus. MATERIALS AND METHODS: This study evaluated the antioxidant and mechanism(s) of inhibitory potential of aqueous extract of C. esculentus on α-amylase and α-glucosidase in vitro. The extract was investigated for its radical scavenging and hypoglycaemic potentials using standard experimental procedures. Lineweaver-Burke plot was used to predict the manner in which the enzymes were inhibited. RESULTS: The data obtained revealed that the extract moderately and potently inhibited the specific activities of α-amylase and α-glucosidase, respectively. The inhibition was concentration-related with respective IC50 values of 5.19 and 0.78 mg/mL relative to that of the control (3.72 and 3.55 mg/mL). The extract also significantly scavenged free radicals and the effects elicited could be ascribed to its phytoconstituents. CONCLUSION: The respective competitive and non-competitive mode of action of the extract is due to its inhibitory potentials on the activities of α-amylase and α-glucosidase. Going forward, in addition to completely characterize the exact compound(s) responsible for the elicited activity in this study, pertinent attention will be given to the in vivo evaluation of the identified constituents.

Time-kill kinetics and biocidal effect of Euclea crispa leaf extracts against microbial membrane.

OBJECTIVE: To evaluate antimicrobial potential of the fractions partitioned from Euclea crispa leaf extract and determination of their impact on cell membrane disruption. METHODS: Antimicrobial potentials were evaluated via susceptibility test, determination of minimum inhibitory concentrations (MICs) and time-kill kinetics of the potent fractions. Degree of membrane disruption was determined by the amount of proteins and nucleotides released from within the cells and SEM images of the membrane after 120 min of treatment. RESULTS: The largest inhibition zone (25.5 ± 0.50 mm) was obtained by ethylacetate fraction against Aeromonas hydrophilla at 10 mg/mL. The lowest MIC (0.16 mg/mL) was exhibited by n-butanol and ethylacetate fractions against test bacteria while all fractions exhibited MIC values between 0.31 and 1.25 mg/mL against susceptible yeast. n-Butanol fraction achieved absolute mortality against Bacillus pumulis (B. pumulis) and Klebsiella pneumoniae (K. pneumoniae) after 90 and 120 min contact time respectively at 1 × MIC. Total mortality also achieved by n-hexane fraction against B. pumulis and K. pneumoniae after 90 and 120 min respectively at 2 × MIC. Ethylacetate fraction achieved absolute mortality against both bacteria after 120 min at 2 × MIC. n-Hexane fraction achieved total mortality against Candida albicans after 120 min at 1 × MIC. Maximum amount of proteins (0.566 µg/mL) was released from K. pneumoniae by n-butanol fraction at 2 × MIC after 120 min of treatment while the maximum amount of nucleotides released (4.575 µg) was from B. pumulis by n-hexane fraction under similar condition. CONCLUSION: This study suggests the leaf of Euclea crispa a source of bioactive compound with membrane attack as one of the mechanisms of its biocidal action.

Antidiabetic Medicinal Plants Used by the Basotho Tribe of Eastern Free State: A Review.

Diabetes mellitus (DM) belongs to the group of five leading important diseases causing death globally and remains a major health problem in Africa. A number of factors such as poverty, poor eating habit, and hormonal imbalance are responsible for the occurrence of the disease. It poses a major health challenge in Africa continent today and the prevalence continues to increase at an alarming rate. Various treatment options particularly the usage of herbs have been effective against diabetes because they have no adverse effects. Interestingly, South Africa, especially the Basotho tribe, is blessed with numerous medicinal plants whose usage in the treatment of DM has been effective since the conventional drugs are expensive and often unaffordable. The present study attempted to update the various scientific evidence on the twenty-three (23) plants originating from different parts of the world but widely used by the Sotho people in the management of DM. Asteraceae topped the list of sixteen (16) plant families and remained the most investigated according to this review. Although limited information was obtained on the antidiabetic activities of these plants, it is however anticipated that government parastatals and scientific communities will pay more attention to these plants in future research.

THE PURVIEW OF PHYTOTHERAPY IN THE MANAGEMENT OF KIDNEY DISORDERS: A SYSTEMATIC REVIEW ON NIGERIA AND SOUTH AFRICA.

BACKGROUND: The kidney is tasked with a number of metabolic functions in the body. In its role as a detoxifier and primary eliminator of xenobiotics, it becomes vulnerable to developing injuries. Currently, over 1 million people in the world are living on renal replacement therapies (RRTs). The case in sub-Sahara African countries like Nigeria and South Africa is not any better than the global trend. MATERIALS AND METHODS: A systematic review of medicinal plants used in the treatment of kidney disorders was conducted. Information were gathered from published scientific journals, books, reports from national, regional and international institutions, conference proceedings and other high profile intellectual resources. MeSH words like 'prevalence of kidney disorders in Africa', 'renal replacement therapy', 'nephrotoxins or nephrotoxicants', 'nephroprotective plants', 'nephroprotective plants in Nigeria or South Africa' and 'nephroprotective phytocompounds' were used to retrieve information from online databases (Google, Pubmed, MEDLINE, Science Direct, Scopus and SID). RESULTS: Interestingly, our findings revealed that phytotherapy has emerged and is being employed to protect renal functions and delay progression of renal pathological conditions into end episodes where the last resort is RRT. In fact, in recent times, Phytotherapists are not only interested in developing relatively safe, more affordable, easily accessible and potent nephroprotective formulations but also increasing awareness on the prevalence of the disease and educating the populace on the probable preventive measures. More importantly, efforts at scientifically elucidating the pharmacological efficacy of the identified nephroprotective plants yet to be validated must be intensified through informed expert opinions. Till date, there is paucity of information on the concept of nephroprotection in most developing countries where kidney disorder is a major threat. Although, the concept is just emerging in South Africa, evidences have given credence to its application in complementary and alternative system of medicine in Nigeria. CONCLUSION: This review, therefore, reawaken researchers' consciousness in the continuous search for auspicious nephroprotective plants that could potentially be excellent candidates in developing new lead drugs to manage and treat renal disorders.

Toxicopathological Evaluation of Hydroethanol Extract of Dianthus basuticus in Wistar Rats.

Background. Dianthus basuticus is a commonly used medicinal plant in Basotho traditional medicine for the treatment of diabetes, but there is no report on its safety or toxicity. Therefore, we evaluated the toxicity profile of the hydroethanol whole plant extract of Dianthus basuticus in Wistar rats. Methods. Acute toxicity test was performed with single oral administration of 100-3200 mg/kg body weight of D. basuticus extract to rats and the animals were observed for 14 days for signs of toxicity. The subacute toxicity experiment was conducted by oral administration of graded doses (200, 400, and 800 mg/kg) of D. basuticus extract daily for 28 days. Behavioural changes as well as haematological, biochemical, and histological parameters were then evaluated. Results. There was no observable sign of toxicity in the acute toxicity test. There were significant decreases (P < 0.05) in the feed and water intake as well as total cholesterol and triglycerides of the D. basuticus extract-treated rats in subacute toxicity study. There were no treatment related differences in the haematological, biochemical, and histopathological evaluations. Conclusions. Administration of hydroethanol extract of D. basuticus may be safe at the dosages tested in this study but its continuous usage can cause anorexia.

Protective Effect of Free and Bound Polyphenol Extracts from Ginger (Zingiber officinale Roscoe) on the Hepatic Antioxidant and Some Carbohydrate Metabolizing Enzymes of Streptozotocin-Induced Diabetic Rats.

This study investigated the hepatoprotective effects of polyphenols from Zingiber officinale on streptozotocin-induced diabetic rats by assessing liver antioxidant enzymes, carbohydrate-metabolizing enzymes and liver function indices. Initial oral glucose tolerance test was conducted using 125 mg/kg, 250 mg/kg, and 500 mg/kg body weight of both free and bound polyphenols from Z. officinale. 28 day daily oral administration of 500 mg/kg body weight of free and bound polyphenols from Z. officinale to streptozotocin-induced (50 mg/kg) diabetic rats significantly reduced (P < 0.05) the fasting blood glucose compared to control groups. There was significant increase (P < 0.05) in the antioxidant enzymes activities in the animals treated with both polyphenols. Similarly, the polyphenols normalised the activities of some carbohydrate metabolic enzymes (hexokinase and phosphofructokinase) in the liver of the rats treated with it and significantly reduced (P < 0.05) the activities of liver function enzymes. The results from the present study have shown that both free and bound polyphenols from Z. officinale especially the free polyphenol could ameliorate liver disorders caused by diabetes mellitus in rats. This further validates the use of this species as medicinal herb and spice by the larger population of Nigerians.

Medicinal potential of Morella serata (Lam.) Killick (Myricaceae) root extracts: biological and pharmacological activities.

BACKGROUND: Morella serata is a South African medicinal plant used in the treatment of microbial infections and to enhance male sexual performance. There is dearth of information in scientific literature on its efficacy and safety. METHODS: In the present study, the root extracts were investigated for the phytochemicals that may be present the antibacterial, anticandida activity using 96 wells microtitre plate method and cytotoxicity using brine shrimp (Artemia salina) lethality assay. RESULTS: The qualitative phytochemical screening revealed the presence of tannins, saponins, flavonoids, terpenoids and steroids. All the extracts including water inhibited both Gram positive and Gram negative bacteria strains at minimum inhibitory concentrations (MIC) ranging from 0.09 - 6.25 mgmL-1. The best activity was observed in the acetone extract inhibiting all the bacteria tested at MIC range of 0.09 - 0.78 mgmL-1 except Shigella flexneri KZN that was inhibited at 1.56 mgmL-1. Similarly, all the extracts suppressed the growth of all Candida species and Trichophyton mucoides at MIC ranging from 0.13 - 3.13 mgmL-1. The cytotoxicity assay revealed potent cytotoxic potential of M. serata methanol and ethanol root extracts by displaying LC50 of 0.26 and 0.18 µgmL-1 respectively. CONCLUSION: The results obtained from the present study indicated broad spectrum antimicrobial activity and justifies the use of the plant in the treatment of infectious diseases. Also the species could be a good natural source of antitumor compounds considering its lethality against brine shrimp nauplii.

Toxicity profile of ethanolic extract of Azadirachta indica stem bark in male Wistar rats.

OBJECTIVE: To investigate the toxic implications of ethanolic stem bark extract of Azadirachta indica (A. indica) at 50, 100, 200 and 300 mg/kg body weight in Wistar rats. METHODS: Fifty male rats of Wistar strains were randomly grouped into five (A-E) of ten animals each. Animals in Group A (control) were orally administered 1 mL of distilled water on daily basis for 21 days while those in Groups B-E received same volume of the extract corresponding to 50, 100, 200 and 300 mg/kg body weight. RESULTS: The extract did not significantly (P>0.05) alter the levels of albumin, total protein, red blood cells and factors relating to it whereas the white blood cell, platelets, serum triacylglycerol and high-density lipoprotein cholesterol decreased significantly (P<0.05). In contrast, the final body weights, absolute weights of the liver, kidney, lungs and heart as well as their organ-body weight ratios, serum globulins, total and conjugated bilirubin, serum cholesterol, low-density lipoprotein cholesterol and computed atherogenic index increased significantly. The spleen-body weight ratio, alkaline phosphatase, alanine and aspartate transaminases, sodium, potassium, calcium, feed and water intake were altered at specific doses. CONCLUSIONS: Overall, the alterations in the biochemical parameters of toxicity have consequential effects on the normal functioning of the organs of the animals. Therefore, the ethanolic extract of A. indica stem bark at the doses of 50, 100, 200 and 300 mg/kg body weight may not be completely safe as an oral remedy and should be taken with caution if absolutely necessary.

Evaluation of aqueous extract of Felicia muricata leaves for anti-inflammatory, antinociceptive, and antipyretic activities.

CONTEXT: Felicia muricata Thunb. (Nees) (Asteraceae) leaves are used in folklore medicine of South Africa as an oral remedy for pain and inflammation. However, the efficacy of the plant part is yet to be validated with scientific experiments. OBJECTIVE: The current study is an effort to investigate the anti-inflammatory, antinociceptive, and antipyretic activities of aqueous extract of F. muricata leaves. MATERIALS AND METHODS: The phytochemical screening of aqueous extract of Felicia muricata leaves and the efficacy of the extract at the doses of 50, 100, and 200 mg/kg body weight was investigated in experimental animals using several models of inflammation (paw edema induced by carrageenan and egg albumin), nociception (acetic acid-induced writhing, formalin-induced pain and tail immersion), and fever (brewer's yeast-induced hyperthermia). RESULTS: The extract contained alkaloids, flavonoids, tannins, saponins, and phenolics. The extract dose-dependently reduced (P <0.05) the number of writhes and stretches induced by acetic acid, number of licks induced by formalin, paw volumes induced by carrageenan and egg albumin. The reaction time by the tail of the extract-treated animals to the hot water also increased. The extract also reduced hyperthermia induced by brewer's yeast. The highest dose (200 mg/kg body weight of the extract) produced the best result in all cases. DISCUSSION AND CONCLUSION: This study revealed that the aqueous extract of Felicia muricata leaves possessed anti-inflammatory, antinociceptive and antipyretic activities. These findings have therefore supported the use of aqueous extract of Felicia muricata leaves in the traditional medicine of South Africa as an oral remedy for pains, inflammation, and fever.