Desert date seed extract-loaded chitosan nanoparticles ameliorate hyperglycemia and insulin deficiency through the reduction in oxidative stress and inflammation.

Plants represents a huge source of bioactive materials that have been used since the old times in the treatment of many diseases. Balanites aegyptiaca, known as desert date, has been used in treatment of fever, diabetes and bacterial infection. Desert dates contains a hard seed that resembles 50-60% of the fruit. The seed extract contains many fatty acids, amino acids and other bioactive materials that gives the extract its antioxidant and anti-inflammatory properties. The study aimed to use Balanites seed extract-loaded chitosan nanoparticles (SeEx-C NPs) for the treatment of streptozotocin (STZ)-induced diabetes in male Sprague Dawley rats. Animals were divided into two main divisions (healthy and diabetic rats). Each division contained seven groups (5 rats/group): control untreated group I, SeEx treated group II and group III (10 and 20 mg/kg b.w., respectively), C NPs treated group IV and group V (10 and 20 mg/kg b.w., respectively) and SeEx-C NPs treated group VI and group VII (10 and 20 mg/kg b.w., respectively). The therapeutical effects of SeEx-C NPs were evaluated through biochemical and immunological assessments in rats' pancreases. The results showed that SeEx-C NPs (10 and 20 mg/kg b.w.) reduced the oxidative stress and inflammation in rats' pancreases allowing the islets neogenesis. The loading of SeEx on C NPs allowed the delivery of fatty acids (oleic, lauric and myristic acid), amino acids (lysine, leucine, phenylalanine and valine) and minerals to pancreatic beta-cells in a sustainable manner. SeEx-C NPs administration successfully increased insulin secretion, allowed pancreatic islets neogenesis and reduced oxidative stress and inflammation.

In vivo investigation of the anti-liver fibrosis impact of Balanites aegyptiaca/ chitosan nanoparticles.

Balanites aegyptiaca (B. aegyptiaca) is an African herb with traditional medical applications. Various pathogenic factors cause hepatic fibrosis and require novel treatment alternatives. Nanoformulation-based natural products can overcome the available drug problems by increasing the efficacy of natural products targeting disease markers. The current study investigated B. aegyptiaca methanolic extract using high-pressure liquid chromatography (HPLC), and B. aegyptiaca/chitosan nanoparticles were prepared. In vivo, evaluation tests were performed to assess the curative effect of the successfully prepared B. aegyptiaca/chitosan nanoparticles. For 30 days, the rats were divided into six groups, typical and fibrosis groups, where the liver fibrosis groups received B. aegyptiaca extract, silymarin, chitosan nanoparticles, and B. aegyptiaca/chitosan nanoparticles daily. In the current investigation, phenolic molecules are the major compounds detected in B. aegyptiaca extract. UV showed that the prepared B. aegyptiaca /chitosan nanoparticles had a single peak at 280 nm, a particle size of 35.0 ± 6.0 nm, and a negative charge at - 8.3 mV. The animal studies showed that the synthetic B. aegyptiaca/chitosan nanoparticles showed substantial anti-fibrotic protective effects against CCl4-induced hepatic fibrosis in rats when compared with other groups through optimization of biochemical and oxidative markers, improved histological changes, and modulated the expression of Col1a1, Acta2 and Cxcl9 genes, which manage liver fibrosis. In conclusion, the current research indicated that the prepared B. aegyptiaca/chitosan nanoparticles improved histological structure and significantly enhanced the biochemical and genetic markers of liver fibrosis in an animal model.

Preparation of chitosan nanoparticles loaded with Balanites aegyptiaca extract for treatment of streptozotocin-induced diabetes in rats.

Diabetes mellitus is characterized by elevated blood sugar level due to a deficiency in insulin production and/or action. Balanites aegyptiaca (BA) has been employed as a hypoglycemic medication. Nanoparticles (NPs) have many advantages like minimized drug dose, sustainable drug release, maximized bioavailability and delivery of drugs. The study aimed to synthesize novel chitosan (CS) NPs loaded with BA extract (BA Ex). The prepared NPs were examined in treatment of streptozotocin-induced diabetes in rats. The anti-diabetic efficiency was evaluated through measuring of levels of blood glucose, insulin, lipid profile, oxidative stress markers, pro-inflammatory cytokines. GC-MS, HPLC and ICP techniques showed the presence of numerous bioactive components that have an anti-diabetic effectiveness. BA Ex-CS NPs succeeded in treatment of diabetes; where, it increased insulin secretion, lowered both FBG and FTA levels and helped in neogenesis of pancreatic islets beta cells. The regenerative activity of BA Ex-CS NPs is attributed to its high antioxidant and anti-inflammatory properties. This antioxidant activity scavenged the generated free radicles that resulted from STZ administration. CS NPs raised the plant extract efficacy, prevented its degradation, and regulated the release of its components. The delivery of BA Ex bioactive components has been revolutionized by CS NPs.

Revealing how phenytoin triggers liver damage and the potential protective effects of Balanites Aegyptiaca fruit extracts: Exploring Nrf2/MAPK/ Beclin-1 signaling pathways.

Phenytoin-induced liver injury (PHT ILII) is a serious condition that may necessitate discontinuation of the drug. This study investigates the mechanisms of PHT ILII and evaluates the protective effects of Balanites Aegyptiaca (BA) fruit extracts on the liver. We focus on the Nrf2/MAPK/NF-κB/Beclin-1 signaling pathways involved in oxidative stress and inflammation from drug-induced liver injury. Phytochemical analyses of BA fruit extracts (Bu-F and EA-F) are conducted. Molecular docking techniques explore the interaction between phenytoin (PHT) and the Nrf2/MAPK/NF-κB/Beclin-1 pathways. Thirty-six male rats are divided into Control, Bu-F, EA-F, PHT, Bu-F/PHT, and EA-F/PHT groups, and they are observed for 45 days. EA-F extract is rich in phenolics/flavonoids, while Bu-F extract mainly contains saponins.PHT ILII causes histological damage in liver tissues and affects Nrf-2, MAPK, TNF-α, IL-1ß, Mcp-1, Beclin-1, iNOS expression, and liver function markers (ALT, AST, ALP). However, EA-F/Bu-F extracts effectively improve the histological structure and significantly reduce biochemical/immunohistochemical parameters, restoring them to near-normal levels. EA-F extract is particularly effective.In conclusion, the Nrf2/MAPK /Beclin-1 pathways play a critical role in the development of PHT ILII. BA fruit extracts show promise as hepato-protective agents, with the EA-F extract demonstrating superior efficacy. These results lay the groundwork for new treatments for PHT ILII and drug-induced liver injuries.

BAAE-AgNPs Improve Symptoms of Diabetes in STZ-induced Diabetic Rats.

OBJECTIVES: Nanoparticles can be employed to improve the therapeutic activity of natural products. Type 2 diabetes mellitus is a serious health condition that has spread like a "modern pandemic" worldwide. In the present study, we developed silver nanoparticles, Ag-NPs, with an aqueous extract from Balanites aegyptiaca to investigate their antioxidant and anti-inflammatory activity in STZ-induced diabetic rats. METHODS: Aqueous extracts of Balanites aegyptiaca seeds (BAAE) were used in the synthesis of BAAE-AgNPs, which were characterized using FTIR and TEM. Different doses of BAAE-AgNP (1/50 LD50; 29.4 mg/kg b.w. and 1/20 LD50: 73.5 mg/kg b.w.) were administered to STZ-induced diabetic rats to evaluate their potential antidiabetic activity. RESULTS: FTIR spectral data indicated the presence of flavonoids and polyphenols in BAAEAgNPs. The size of the BAAE-AgNPs, determined by TEM examination, was 49.33 ± 7.59 nm, with a zeta potential of +25.37. BAAE-AgNPs were characterized by an LD50 value of 1470 mg/kg b.w. In diabetic rats, the daily oral administration of both doses of BAAE-AgNPs (29.4 and 73.5 mg/kg b.w.) for 12 weeks resulted in a significant improvement in body weight, insulin homeostasis, HbA1c, HDL-C, MDA, and pancreatic SOD, CAT, and GSH. They reduced plasma glucose, cholesterol, and triglycerides. This treatment also resulted in a significant decrease in pancreatic IL-6, p53, and TNF-α in diabetic rats. Furthermore, BAAE-AgNPs down-regulated pancreatic TGF-ß1 and Akt gene expression in diabetic rats and resulted in a significant decrease in the regulation of hepatic GLUT-2, as well as an increase in the regulation of hepatic GK and pancreatic B-cl2 gene expression. The histopathological results obtained indicated that BAAEAgNPs improved pancreatic tissue metabolism by enhancing antioxidant enzymes, suppressing inflammatory cytokines, and scavenging free radicals. CONCLUSION: The findings implied that similar to the glibenclamide-treated groups, in the BAAEAgNPs treated group, the compromised antioxidant status normalized in STZ-induced diabetes. By scavenging free radicals, BAAE-Ag-NPs protected against lipid peroxidation while reducing the risk of complications from diabetes. Compared to the daily dose of 29.4 mg, the impact was more prominent at 73.5 mg.

Investigation of Potential In Vitro Anticancer and Antimicrobial Activities of Balanites aegyptiaca (L.) Delile Fruit Extract and Its Phytochemical Components.

The therapeutic importance of Balanites aegyptiaca in folk medicine for the treatment of several common human diseases has led researchers to conduct phytochemical and pharmacological studies on extracts from various parts of the plant. In the current study, the phytochemical composition of the B. aegyptiaca methanolic fruit extract was characterized, and its antimicrobial activity was evaluated together with the cytotoxic activity against MCF-7, PC-3, and Caco-2, compared with normal Vero cells. Further, its effects on cell cycle arrest, apoptosis induction and expression of apoptosis-related genes were assessed. The phytochemical screening revealed the presence of fatty acids and their esters in addition to phytosterols, steroid derivatives, and bioflavonoid glycosides with oleic and palmitic acids being the prevalent components (24.12 and 21.56%, respectively). The results showed considerable cytotoxic activity of the extract against the three cancer cell lines (MCF-7, PC-3, and Caco-2) with a selectivity index ranging from 5.07 to 6.52. This effect was further confirmed with the accompanied increased total apoptosis of treated PC-3 cells (19.22% of the total number of cells) compared to the control cells (0.64% of the total number of cells) with cell cycle arrest at G1 phase and the increased transcription of pro-apoptotic genes including P53 (3.69) and BAX (3.33) expressed as fold change (2^ ΔΔCT). The calculated minimum inhibitory concentration (MIC) was similar (62.5 µg/mL) against the three tested bacterial strains (Acinetobacter johnsonii, Serratia marcescens and Agrobacterium tumefaciens), while it was higher than 1000 µg/mL for the fungal species (Rhizoctonia solani, Penicillium italicum, and Fusarium oxysporium). Our findings suggest a promising anticancer activity for B. aegyptiaca, which paves the way for more detailed future studies.

Cardioprotective and Antioxidant Efficiency of Balanites aegyptiaca Extract Against Doxorubicin® Complication.

<b>Background and Objective:</b> The use of Doxorubicin^® (Doxo) in the treatment of different tumours is restricted due to its cardiotoxicity. The objective of this study was to determine the protective effect of<i> Balanites aegyptiaca</i> extract against cardiotoxicity induced by Doxorubicin^® in male rats. <b>Materials and Methods:</b> Adult male rats (140-160) were parted into 6 groups (10 animals each) as follows: Group (1) Normal rats the control, group (2) Rats were administered BAE (200 mg kg¹) orally for 4 weeks, group (3) Rats were treated IP with the anticancer drug (Doxorubicin^®) at the dose of (0.5 mg kg¹) for 4 weeks, group (4) Administrated orally with BAE in combination with Doxo injection for 4 weeks, group (5) Rats orally with BAE before intoxication with Doxo for 4 weeks and finally group (6) Animals post-administration of BAE for 4 weeks after intoxication with Doxo. After 4 weeks of injections. <b>Results:</b> Revealed that BAE succeeded to decline the Doxorubicin cardiotoxicity, this was evidenced by the significant reduction of serum LDH, CK-MB, total cholesterol, triglycerides, HDL, TNF-α, IL-1ß and IL-6 as well as cardiac MDA and nitric oxide levels coupled with marked improvement in serum LDL, PON1 as well as cardiac GSH, SOD and CAT. Moreover, the BAE induced prominent regeneration of the cardiac muscle. <b>Conclusion:</b> <i>Balanites aegyptiaca</i> extract may be a promising cardio-protector against Doxorubicin^® toxicity mediated through their antioxidant and radical scavenging activities.

Provenance effect on bioactive phytochemicals and nutritional and health benefits of the desert date Balanites aegyptiaca.

Balanites aegyptiaca L. is a multipurpose tree distributed in Africa and Middle East. Several parts of B. aegyptiaca have been suggested to have medicinal uses. So far the effect of ecological origin on the nutritional values and biological activities of B. aegyptiaca genotypes is rarely investigated. Further, metabolic profiling and assessment of the functional food value of B. aegyptiaca leaves are far from complete. In this study, biological activities and profiling of primary and secondary metabolites were investigated in the leaves of five B. aegyptiaca provenances collected from Egypt, Sudan, Saudi Arabia, and Yemen. Interestingly, all provenances showed notable antidiabetic, antioxidant, antiprotozoal, antibacterial, antifungal, and anticancer activities. Hierarchical clustering analysis revealed significant variability in the concentrations of individual sugars, organic acids, amino acids, fatty acids, vitamins, phenolics, and minerals among the provenances and these variations were provenance dependent. Medina provenance showed the heights diphenylpicrylhydrazyl (DPPH) scavenging and antifungal activities and was the most powerful against embryonic kidney adenocarcinoma and urinary bladder carcinoma cells. The highest inhibition against Escherichia coli and colon carcinoma cells was observed by Sudan and Cairo provenances. El-Kharga and Yemen provenances showed the greatest activity against Trypanosoma cruzi and hepatocellular and urinary bladder carcinoma. Therefore, leaves of B. aegyptiaca possess good nutritive and biological capacities and might have potential applications in the food and medical industries. However, the strength of such activities is significantly affected by the provenance. PRACTICAL APPLICATIONS: According to the national Research Council (NRC) of United States, Balanites aegyptiaca L. is recognized among the 24 priority lost crops of Africa. B. aegyptiaca leaves contain considerable amounts of primary metabolites (e.g., sugars, EAAs, USFAs) and secondary (e.g., phenolic acids and flavonoids) metabolites, vitamins, and macro and microelements. The obvious existence of these nutritionally and medicinally related compounds supports the functional food value of B. aegyptiaca leaves. Moreover, the present results revealed that B. aegyptiaca is not only a foliage dietary plant, but also could be considered as a valuable source for neutraceuticals, which support its pharmacological value. So far, this is the first report to explore, in detail, the functional food value of B. aegyptiaca leaves by presenting a clear image about its metabolic profiling and biological activities, and how the provenance factor could affect these values.

Phytochemicals, Antioxidant Activity and Ethnobotanical Uses of Balanites aegyptiaca (L.) Del. Fruits from the Arid Zone of Mauritania, Northwest Africa.

Phytochemicals and antioxidant activity of fruits of 30 B. aegyptiaca trees naturally growing in the hyper-arid and arid zones in Mauritania were evaluated by following standard procedures. Ethnobotanical uses of fruit pulps and kernel were assessed using a structured questionnaire. Balanites aegyptiaca fruit pulp is a good source of sugars (33 g/100 g dry matter (DM)), polyphenols (264 mg GAE/100 g DM) and flavonoids (34.2 mg/100 g DM) with an average antioxidant activity of 519 µmol TEAC/100 g DM. The fruit kernel is rich in lipids (46.2 g/100 g DM) and proteins (29.5 g/100 g DM). Fruits from the hyper-arid zone exhibited high level of polyphenols, antioxidant activity and soluble tannins. Almost all of the informants (97.14%) reported the use of fruit pulp in folk medicine to treat diabetes, while 72.86% reported using the fruit pulp to treat hypertension. Kernel oil is mainly employed as ointments in the treatment of paronychia (57.14%) and dermal infections (35.71%). The predominant methods for preparing/administering fruit pulp/Kernel were maceration (58.8%), sucking fruit pulp (25.7%) and decoction (24.2%). Balanites aegyptiaca fruit contain both nutritional and health-promoting phytochemicals that could be of interest in the development of strategies for sustainable use of this neglected indigenous fruit tree.

Evaluation of the antiulcer activity of the aqueous stem bark extract of Balanites aegyptiaca L Delile in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Balanites aegyptiaca L. Delile (Zygophyllaceae) is a perennial tree that is mostly found in Africa, south Asia and most desert environments. Decoctions of its stem barks are used in northern Nigeria for the treatment of ulcers and stomach issues. Other folkloric uses include treatment of wounds, malaria, dysentery, asthma, and fever. AIM OF THE STUDY: The present study evaluated the antiulcer activity of the aqueous stem bark extract of Balanites aegyptiaca in Wistar rats. MATERIALS AND METHODS: The antiulcer activity of the aqueous stem bark extract of Balanites aegyptiaca (125, 250 and 500 mg/kg, p.o.) was evaluated in ethanol, indomethacin, pylorus ligation and acetic acid-induced ulcer models in rats. Parameters such as mean ulcer indices and percentage ulcer inhibition were assessed in ethanol, indomethacin and acetic acid-induced ulcer models while gastric volume, pH, and titratable acidity were evaluated in the pylorus ligation ulcer model. RESULTS: The extract at the doses of 125, 250 and 500 mg/kg caused a significant (p < 0.01), dose dependent reduction in mean ulcer indices in the ethanol and indomethacin ulcer models. A significant dose dependent reduction in mean ulcer indices were also observed after three (p < 0.01) and seven (p < 0.001) days of treatment with the extract in acetic acid-induced ulcer model. In pylorus ligation model, the gastric secretion parameters (gastric volume, pH, and titratable acidity) showed no alteration in the different doses of the extract when compared to the control. CONCLUSION: Our study showed that the aqueous stem bark extract of Balanites aegyptiaca possesses gastroprotective and ulcer healing properties and therefore not only provides scientific evidence for its folkloric use in the treatment of ulcers but also showed evidence that it may be used in the development of a new phytotherapeutic formulation for the treatment of peptic ulcer.

Balanites aegyptiaca ameliorates insulin secretion and decreases pancreatic apoptosis in diabetic rats: Role of SAPK/JNK pathway.

SAPK-JNK pathway performs a significant role in the pathogenesis of type 2 diabetes. Balanites aegyptiaca (BA) is used as an anti-diabetic agent in folk medicine however its hypoglycemic mechanism is not fully elucidated. The current study aimed to evaluate the effect of crude extract, butanol, and dichloromethane fractions from BA on the stress-activated protein kinase/c-Jun N-terminal kinase (SAPK-JNK) pathway in experimental diabetic rats. Six groups of male Wistar rats were included: normal control, diabetic, diabetic rats treated with crude, butanol or dichloromethane fraction from BA (50 mg/kg BW) and diabetic rats treated with gliclazide as a reference drug for one month. Our results suggested a protective role of treatment of diabetic rats with BA against oxidative stress-induced SAPK-JNK pathway. Moreover, BA treatment produced a reduction in plasma glucose, HbA1c, lactic acid, lipid profile, malondialdehyde levels and produced an increase in insulin, reduced glutathione levels, catalase and superoxide dismutase activities compared with untreated diabetic rats. Moreover, it decreased apoptosis signal-regulating kinase 1, c-Jun N-terminal kinase 1, protein 53 and increased insulin receptor substrate 1 in rat pancreas while it increased glucose transporter 4 in rat muscle. Analysis of BA extracts by LC-HRMS revealed the presence of different saponins with reported hypoglycemic effect. In conclusion, BA exerted hypoglycemic, hypolipidemic, insulinotropic and antioxidant effects. Additionally, it reduced apoptosis in pancreatic ß-cells and increased glucose uptake in muscle. These results suggest that the hypoglycemic effect of BA is due to the inhibition of the SAPK-JNK pathway.

Randomized double-blinded pilot clinical study of the antidiabetic activity of Balanites aegyptiaca and UPLC-ESI-MS/MS identification of its metabolites.

CONTEXT: Balanites aegyptiaca Del. (Zygophyllaceae) fruits are traditionally known for the treatment of hyperglycaemia. Several in vitro and in vivo studies proposed some mechanisms of action. However, clinical trials in human beings were never reported to date. OBJECTIVES: To investigate the antidiabetic efficacy of the 70% ethanol extract of the pericarps of B. aegyptiaca (BE) within a nutritional intervention in elderly people. MATERIALS AND METHODS: Ultra-performance electrospray ionization-mass spectroscopy (UPLC-ESI-MS/MS) analysis was used for metabolic profiling of BE which was incorporated in hard gelatine capsules (400 mg/day) and tested on 30 type 2 diabetes (T2D) Egyptian patients for 8 weeks. According to sex, age and body mass index participants were divided into two equivalent groups, placebo and treatment. RESULTS: Thirteen compounds were identified in BE using UPLC-ESI-MS/MS analysis among which five steroidal saponins, seven phenolic compounds and a sterol glucoside. At the end of the 8-week treatment, the treated group showed 26.88% decrease in 2 h postprandial plasma glucose relative to 2.6% increase in the placebo group, while fasting plasma glucose was reduced to 10.3%. Treatment with BE capsules for 8 weeks produced significant reduction in the plasma triglyceride, total cholesterol and low-density lipoprotein cholesterol by 9.0, 12.76 and 21.35%, respectively, with 29.8% increase in the high-density lipoprotein cholesterol. Plasma alanine transaminase and aspartate transaminase were reduced by 42.6 and 43.3%, respectively. DISCUSSION AND CONCLUSION: Administration of the BE capsules to T2D resulted in significant improvements in the glycaemic markers and the lipid profile, without adverse effects or hypoglycaemia.

Management of Hyperglycaemia by Ethyl Acetate Extract of Balanites aegyptiaca (Desert Date).

Reactive oxygen species play a significant role in the pathogenesis of retinopathy in diabetes patients. The current study aimed to assess the effect of ethyl acetate extract (EAE) from Balanites aegyptiaca (10, 25 or 50 mg/kg b.w.) in experimental diabetic rats. To achieve this aim, five groups of male rats were included: control, diabetic, and diabetic rats treated with 10, 25, and 50 µg/kg b.w. of EAE for eight weeks. Our results suggests a protective role of EAE against oxidative stress induced by streptozocine. EAE treatment produced a reduction in blood glucose levels, HbA1c, malondialdehyde and vascular endothelial growth factor (VEGF) in diabetic retina (p < 0.001), as well as an enhancement in antioxidant capacity against streptozocine-induced oxidative stress. Tumor necrosis factor alpha (TNF-α), interleukin (IL-1ß) and vascular endothelial growth factor (VEGF) were significantly reduced in diabetic rats treated with EAE, compared with untreated diabetic rats. Analysis of EAE by GC-MS indicated the presence of ß-sistosterol. Overall, EAE modulates oxidative stress induced by streptozocine and enhances antioxidant activity, which may provide additional endothelial protection in retina of diabetic rats. These results hold great promise in the management of diabetic complications.

Unraveling the active hypoglycemic agent trigonelline in Balanites aegyptiaca date fruit using metabolite fingerprinting by NMR.

Trigonelline (3-carboxy-1-methyl pyridinium) was identified as a relevant bioactivity and taste imparting component in Balanites aegyptiaca fruit, using (1)H NMR of crude extracts without any fractionation or isolation step. The structural integrity of trigonelline was established within the extract matrix via(1)H NMR, (1)H-(1)H COSY, HMQC and HMBC and by comparison with authentic standard. A quantitative (1)H NMR method (qHNMR) was used to determine trigonelline concentrations in the peel and pulp of B. aegyptiaca fruit of 8 and 13mgg(-1), respectively. Trigonelline so far has not been reported from B. aegyptiaca or its genus as it easily escapes LC-MS based detection. Its discovery provides novel insight into the balanite fruits antidiabetic properties as the compound is known for a pronounced hypoglycemic effect. In addition, it is likely to impart the perceptible bitter taste portion to balanites sweet bitter taste. UPLC-MS of the crude extract additionally revealed the fruit flavonoid pattern showing quercetin/isorhamnetin flavonol conjugates in addition to epicatechin, the latter being present at much lower levels.

Aldose reductase inhibition of a saponin-rich fraction and new furostanol saponin derivatives from Balanites aegyptiaca.

BACKGROUND: Balanites aegyptiaca Del. (Zygophyllaceae) fruits are used to treat hyperglycemia in Egyptian folk medicine and are sold by herbalists in the Egyptian open market for this purpose. Nevertheless, the fruits have not yet been incorporated into pharmaceutical dosage forms. The identity of the bioactive compounds and their possible mechanisms of action were not well understood until now. PURPOSE: Aldose reductase inhibitors are considered vital therapeutic and preventive agents to address complications caused by hyperglycemia. The present study was carried out to identify the primary compounds responsible for the aldose reductase inhibitory activity of Balanites aegyptiaca fruits. STUDY DESIGN: The 70% ethanolic extract of Balanites aegyptiaca fruit mesocarp and its fractions were screened for inhibition of the aldose reductase enzyme. Bio-guided fractionation of the active butanol fraction was performed and the primary compounds present in the saponin-rich fraction (D), which were responsible for the inhibitory activity, were characterized. HPLC chromatographic profiles were established for the different fractions, using the isolated compounds as biomarkers. METHODS: Aldose reductase inhibition was tested in vitro on rat liver homogenate. The butanol fraction of the 70% ethanolic extract was fractionated using vacuum liquid chromatography (VLC, RP-18 column). The most active sub-fraction D, which was eluted with 75% methanol, was subjected to preparative HPLC to isolate the bioactive compounds. RESULTS: The butanol fraction displayed inhibitory activity against the aldose reductase enzyme (IC50 = 55.0 ± 6 µg/ml). Sub-fraction D exhibited the highest inhibitory activity (IC50 = 12.8 ± 1 µg/ml). Five new steroidal saponin derivatives were isolated from this fraction. The isolated compounds were identified as compound 1a/b, a 7:3 mixture of the 25R:25S epimers of 26-O-ß-D-glucopyranosyl-furost-5-ene-3,22,26-triol 3-O-[α-L-rhamnopyranosyl-(1→3)- ß-D-glucopyranosyl-(1→2)]- α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranoside; compound 2, 26-O-ß-D-glucopyranosyl-(25R)-furost-5-ene-3,22,26-triol 3-O-[ ß-D-glucopyranosyl-(1→2)]- α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranoside; compound 3, 26-O-ß-D-glucopyranosyl-(25R)-furost-5,20-diene-3,26-diol 3-O-[α-L-rhamnopyranosyl-(1→3)- ß-D-glucopyranosyl-(1→2)]- α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranoside; compound 4, 26-O-ß-D-glucopyranosyl-(25R)-furost-5,20-diene-3,26-diol 3-O-[ ß-D-glucopyranosyl-(1→2)]- α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranoside; and compound 5, which is the 25S epimer of compound 4, by using various spectroscopic methods [MS,1D and 2D NMR (HSQC, HMBC, DQF-COSY, HSQC-TOCSY)]. Compounds 1a/b, 2, 3, 4, 5 exhibited highly significant aldose reductase inhibitory activities (IC50 values were 1.9 ± 0.2, 1.3 ± 0.5, 5.6 ± 0.2, 5.1 ± 0.4, 5.1 ± 0.6 µM, respectively) as compared to the activity of the reference standard quercetin (IC50 = 6.6 ± 0.3 µM). CONCLUSION: The aldose reductase inhibitory activity of Balanites fruits is due to the steroidal saponins present. HPLC chromatographic profiles of the crude butanol fraction and its 4 sub-fractions showed that the most highly bioactive fraction D contained the highest amount of steroidal saponins (75%) as compared to the 21% present in the original butanol fraction. The isolated furostanol saponins proved to be highly active in an in vitro assay.

Effects of hydroethanolic extracts of Balanites aegyptiaca (L.) Delile (Balanitaceae) on some resistant pathogens bacteria isolated from wounds.

BACKGROUND: The bark of Balanites aegyptiaca (L.) Delile (Balanitaceae) is widely used in the Togolese folk medicine for the treatment of wounds, malaria and skin diseases. This study aimed to evaluate the antioxidant properties and the activity of the bark extract of this plant against some resistant bacteria isolated from wounds. MATERIALS AND METHODS: The antimicrobial activity was assayed using the NCCLS microdilution while the DPPH free radical scavenging and ferric reducing antioxidant power (FRAP) were used for the antioxidant activity. The susceptibility to conventional antibiotics of bacteria was assessed by an agar disk diffusion method using commercial disk. Phenolics were quantified using spectrophotometric methods. RESULTS: The hydroethanolic extracts of the bark inhibited in vitro the growth of Pseudomonas aeruginosa and Staphylococcus aureus in a dose-dependent manner. Bacteriostatic effects were observed on 15 (33%) and 21 (47%) strains of Staphylococcus aureus and Pseudomonas aeruginosa respectively. The MICs and MBCs against Staphylococcus aureus strains ranged respectively from 2.5 to 12.5 µg/mL and 2.5 to 50 µg/mL. Concerning Pseudomonas aeruginosa, these values were equal (12.5 µg/mL). Multidrug resistance was 95% against six antibiotics for Pseudomonas aeruginosa and 91% against five antibiotics for Staphylococcus aureus. The DPPH radical scavenging and the FRAP assays yielded weak antioxidant activity, closely linked with the total phenolic contents. CONCLUSION: This study indicated that the bark extract of Balanites aegyptiaca possess bioactive compounds implicated in the free radical scavenging and antibacterial activities, justifying the use of the plant in the traditional medicines.

Laboratory assessment of the molluscicidal and cercariacidal activities of Balanites aegyptiaca.

OBJECTIVE: To assess the molluscicidal and cercariacidal activities of aqueous extracts of Balanites aegyptiaca (B. aegyptiaca) against Ethiopian Biomphalaria pfeifferi (B. pfeifferi), Lymnaea natalensis (L. natalensis) and Schistosoma mansoni (S. mansoni) cercariae. METHODS: Extracts of seeds, endocarp, mesocarp, and fruit of B. aegyptiaca were tested for their activities against adult B. pfeifferi and L. natalensis. The cercariacidal activity of the seeds of the plant was also evaluated against S. mansoni. Bioassays were carried out following the methods recommended by WHO. Snail mortalities were compared between each plant part and snail species, and LC50 and LC90 values for the plant parts tested were computed. The cercariacidal activity of the plant was assessed by exposing the mice to the cercariae pre-exposed to aqueous extract of B. aegyptiaca seeds. RESULTS: For the molluscicidal activities of seeds, endocarp, mesocarp and whole fruit, the LC50 values against B. pfeifferi were 56.32, 77.53, 65.51 and 66.63 mg/L, respectively, while the respective LC90 values were 77.70, 120.04, 89.50 and 97.55 mg/L. Similarly, the LC50 values for the seeds, endocarp, mesocarp and whole fruit against L. natalensis were 80.33, 92.61, 83.52 and 87.84 mg/L, respectively, while the respective LC90 values were 102.30, 138.21, 115.42 and 127.69 mg/L. B. pfeifferi were found to be more susceptible to B. aegyptiaca than L. natalensis. S. mansoni cercariae exposed to 15 mg/L of extract of seeds were incapable of infecting mice. The mean egg load of tissue was reduced in mice infected with the cercariae exposed to 5 and 10 mg/L of the extract. CONCLUSIONS: The aqueous extracts of different parts of B. aegyptiaca exhibited reasonable molluscicidal activity against B. pfeifferi and L. natalensis, as well as cercariacidal activity against S. mansoni cercariae. However, comprehensive laboratory evaluation is recommended prior to field tests of the plant parts since their impact on other aquatic biota is not known.