Vetiver aerial parts and roots ameliorate rheumatoid arthritis in complete Freund's adjuvant rat model, a phytochemical profiling and mechanistic study.

ETHNOPHARMACOLOGICAL RELEVANCE: Vetiver (Chrysopogon zizanioides) is indigenous to India where it is traditionally used to relief rheumatisms, lumbagos and sprains. Vetiver anti-inflammatory activity has not been previously investigated, and its specific interactions with body inflammation cascade remain largely unknown. AIM OF THE STUDY: The present work was performed to validate the ethnobotanical use of the plant and compare the anti-inflammatory activities of the ethanolic extracts of the most traditionally used part (aerial part) to that of the root. Furthermore, we attempt to reveal the molecular mechanism of this anti-inflammatory activity in correlation to the chemical composition of C. zizanioides aerial (CA) and root parts (CR). MATERIALS AND METHODS: Ultraperformance liquid chromatography coupled to high resolution mass spectrometry (UHPLC/HRMS) was used for comprehensive analysis of both CA and CR. The anti-inflammatory effect of both extracts was evaluated in complete Freund's adjuvant (CFA)-induced RA model in Wistar rats. RESULTS: Phenolic metabolites were predominant in CA and 42 were identified for the first time, while only 13 were identified in CR. Meanwhile, triterpenes and sesquiterpenes were confined to the root extract. In CFA arthritis model, CA showed better anti-inflammatory activity than CR marked by an increase in serum level of IL-10 with simultaneous decrease in pro-inflammatory markers; IL-6, ACPA and TNF-α and was evident in histopathological examination. This anti-inflammatory effect was accompanied by down-regulation of JAK2/STAT3/SOCs3, ERK1/ERK2, TRAF6/c-FOS/NFATC1, TRAF6/NF-κB/NFATC1 and RANKL pathways which were all upregulated after CFA injection. These pathways were modulated to larger extent by CA, except for ERK1/ERK2 which was downregulated more effectively by CR. This differential effect between CA and CR can be explained by the variability in their phytoconstituents profile. CONCLUSION: In agreement with the ethnobotanical preference, CA extract was more effective than CR extract in reducing the symptoms of RA probably due to its enrichment with flavonoids, lignans, and flavolignans. Both CA and CR reduced the production of inflammatory cytokines through modulating various biological signaling pathways. These findings support the traditional use of vetiver leaves as a remedy for RA and suggest that the use of the whole plant may offer advantage by synergistically affecting more inflammatory pathways.

Detailed metabolite profiling and in vitro studies of Urospermum picroides as a potential functional food.

Wild edible plants (WEP) are part of the Mediterranean culinary culture and can be used as famine foods in times of severe food shortages. Urospermum picroides is a WEP that grows under harsh conditions and represents an opportunity to expand and diversify the global food supply. However, little is known about its chemical profile. In this study, liquid chromatography coupled to HRESIMS allowed the identification of 77 metabolites in U. picroides extract, among which 12 sesquiterpene-amino acid conjugates are reported here for the first time. Due to the novelty of these conjugates, GNPS molecular networking was used to provide information on their fragmentation pathway. Further, the sesquiterpene enriched U. picroides extract showed a moderate anti-inflammatory effect in LPS-stimulated THP1-macrophages by increasing IL-10 secretion while decreasing pro-inflammatory IL-6 secretion at 50 µg/mL. Our study provides evidence for the potential use of U. picroides as an anti-inflammatory functional food and nutraceutical agent.

A Proposed Association between Improving Energy Metabolism of HepG2 Cells by Plant Extracts and Increasing Their Sensitivity to Doxorubicin.

Increasing cancer cell sensitivity to chemotherapy by amending aberrant metabolism using plant extracts represents a promising strategy to lower chemotherapy doses while retaining the same therapeutic outcome. Here, we incubated HepG2 cells with four plant extracts that were selected based on an earlier assessment of their cytotoxicity, viz asparagus, green tea, rue, and avocado, separately, before treatment with doxorubicin. MTT assays elucidated a significant decrease in doxorubicin-IC50 following HepG2 incubation with each extract, albeit to a variable extent. The investigated extract's ultra-performance liquid chromatography and gas chromatography coupled with mass spectrometry (UPLC/MS and GC/MS) revealed several constituents with anticancer activity. Biochemical investigation displayed several favorable effects, including the inhibition of hypoxia-inducible factor1α (HIF1α), c-Myc, pyruvate kinase-M2 (PKM2), lactate dehydrogenase-A (LDH-A), glucose-6-phosphate dehydrogenase (G6PD), and glutaminase by asparagus and rue extracts. To less extent, HIF1α, c-Myc, PKM2, and LDH-A were partially inhibited by green tea extract, and HIF1α and glutaminase activity was inhibited by avocado oil. Undesirably, green tea extract increased glutaminase; avocado oil rose c-Myc, and both increased G6PD. In conclusion, our study confirms the potential cytotoxic effects of these plant extracts. It highlights a strong association between the ability of asparagus, green tea, rue, and avocado to sensitize HepG2 cells to doxorubicin and their power to amend cell metabolism, suggesting their use as add-on agents that might aid in clinically lowering the doxorubicin dose.

Metabolic profiling of Ochradenus baccatus Delile. utilizing UHPLC-HRESIMS in relation to the in vitro biological investigations.

Ochradenus baccatus Delile (Resedaceae) is a desert plant with edible fruits native to the Middle East. Few investigators have reported antibacterial, antiparasitic and anti-cancer activities of the plant. Herein we evaluated the cytotoxic activity of O. baccatus using four cell lines and a zebrafish embryo model. Additionally, liquid chromatography coupled with mass spectroscopy was performed to characterize the extract's main constituents. The highest cytotoxicity was observed against human cervical adenocarcinoma (HeLa), with CC50 of 39.1 µg/mL and a selectivity index (SI) of 7.23 (p < 0.01). Metabolic analysis of the extract resulted in the annotation of 57 metabolites, including fatty acids, flavonoids, glucosinolates, nitrile glycosides, in addition to organic acids. The extract showed an abundance of hydroxylated fatty acids (16 peaks). Further, 3 nitrile glycosides have been identified for the first time in Ochradenus sp., in addition to 2 glucosinolates. These identified phytochemicals may partially explain the cytotoxic activity of the extract. We propose O. baccatus as a possible safe food source for further utilization to partially contribute to the increasing food demand specially in Saharan countries.

The phloroglucinol calcitrinone A, a novel mitochondria-targeting agent, induces cell death in breast cancer cells.

Breast cancer is the most common cancer and the leading cause of cancer-related mortality among females worldwide. From the leaves of Callistemon citrinus, we have isolated a novel phloroglucinol dimer, calcitrinone A, and analyzed its potential anticancer activity using the triple-negative breast cancer cell line MDA-MB-231. Calcitrinone A decreased the total intracellular ATP levels, inhibited proliferation, and induced apoptosis in MDA-MB-231 cells, but was less toxic to peripheral blood mononuclear cells. The antiproliferative and apoptosis-inducing effects of calcitrinone A were confirmed in vivo using breast cancer xenografts grown on chick chorioallantoic membranes. Mechanistic analysis showed mitochondrial membrane-potential dissipation and interference with energy-yielding processes resulting in cell accumulation in the S phase of the cell cycle. Seahorse assay analysis revealed an early inhibition of mitochondrial oxidative phosphorylation (OXPHOS). At the molecular level, calcitrinone A inhibited activity of the succinate-coenzyme Q reductase (SQR) (mitochondrial complex II). In silico docking identified the coenzyme Q binding pocket as a possible high affinity binding site for calcitrinone A in SQR. Inhibition of complex II was accompanied by strong elevation of mitochondrial superoxide and cytoplasmic ROS. Calcitrinone A might be a promising anticancer lead compound acting through the interference with the mitochondrial complex II activity.

Oral and Topical Anti-Inflammatory Activity of Jatropha integerrima Leaves Extract in Relation to Its Metabolite Profile.

Jatropha integerrima Jacq., family: Euphorbiaceae, is used in India and subtropical Africa to treat different skin conditions. In this study we evaluated the anti-inflammatory activity of J. integerrima leaves extract (JILE) using rat paw edema model. The extract was administered orally (200 and 400 mg/kg) or applied topically as creams at 2.5, 5, and 10% strength. Four hours post-treatment, maximum reduction of edema volume by 63.09% was observed after oral administration of JILE (400 mg/kg) as compared to indomethacin with 60.43%. The extract anti-inflammatory effect was accompanied by a decrease in NO, prostaglandin PGE2, TNF-α and PKC levels by 19, 29.35, 16.9, and 47.83%, respectively. Additionally, topical applications of JILE showed dose dependent reduction in paw edema and resulted in normalized levels of PGE2, TNF-α, and PKC when used as 10% cream. Signs of inflammations were reduced or absent from paw tissue of animals receiving JILE either orally or topically. Finally, liquid chromatography/mass spectrometry analysis of JILE resulted in the annotation of 133 metabolites including 24 diterpenoids, 19 flavonoids, 10 phenolic acid conjugates, 8 cyclic peptides, 6 phytosterols, 4 sesquiterpenes, and 4 coumarins. Several of the annotated metabolites have known anti-inflammatory activity including vitexin, isovitexin, fraxitin, scopeltin, stigmasterol, and many diterpenoidal derivatives.

Non-polar metabolites of green beans (Phaseolus vulgaris L.) potentiate the antidiabetic activity of mesenchymal stem cells in streptozotocin-induced diabetes in rats.

Green beans (Phaseolus vulgaris L.) are consumed as pods or mature seeds (common beans). The pods were extracted with 95% ethanol and processed to prepare non-polar and polar fractions. Comparing the antihyperglycemic activity of both fractions, non-polar fraction (NPF, 200 mg kg-1  day-1 ) lowered blood glucose in streptozotocin diabetic rats by 65% compared to 57% for the polar fraction at the same dose. When NPF treatment was combined with injection of mesenchymal stem cells (MSC) a 4.4-fold increase in serum insulin and a 73.6% reduction in blood glucose were observed compared to untreated control. Additionally, a significant decrease in malondialdehyde (76.2%), nitric oxide (68.2%), cholesterol (76.1%), and triglycerides (69.5%) and a 1.75-fold increase in HDL concentrations were observed in the group treated with this combination compared to diabetic animals. Interestingly, NPF increased homing of MSC in pancreas potentiating their antidiabetic activity. Finally, 26 compounds were identified in NPF using LC/MS analysis and four were isolated in pure form. The isolated compounds namely calotroproceryl acetate, fridelin, calotroproceryl A, and stigmasterol showed good inhibitory activity against pancreatic lipase with IC50 at 1.93, 1.07, 1.34 and 1.44-1 µg/ml, respectively. Additionally, these compounds inhibited α-amylase, albeit at higher concentration, with IC50 at 248, 212, 254, and 155 µg/ml for calotroproceryl acetate, fridelin, calotroproceryl A, and stigmasterol, respectively. Our results suggest that green beans extract can potentiate effect of MSC in diabetes directly due to its own antidiabetic effect and indirectly by increasing MSC homing in pancreatic tissues. PRACTICAL APPLICATIONS: It has been suggested in this study that green beans can improve hyperglycemia, oxidative balance in diabetes, so green beans can be promoted as a healthy nutrient for diabetic patients. Green beans also can enhance homing and differentiation of mesnchymal stem cells in the pancreas for future stem cell therapy of type I diabetes.

Phytochemical constituents and anti-tyrosinase activity of Macadamia integrifolia leaves extract.

Macadamia integrifolia Maiden & Betche is cultivated around the world for its highly valued nuts (macadamia nuts). Although the chemical composition of the edible macadamia oil has been repeatedly investigated, other plant organs have not been phytochemically or biologically assessed. In this study, ethanolic extract of M. integrifolia leaves was phytochemically investigated which led to the isolation of 6 compounds. Two functional galactolipids, i.e., monogalactosyl diacylglycrol 36:4 (MGDG 36:4), digalactosyl monoacylglycerol 18:2 (DGMG 18:2), gallic acid and protocatechuic acid were identified in the genus Macadamia for the first time, in addition to the cyanogenic glycoside dhurrin and ß-sitosterol. Additionally, anti-tyrosinase activity of the extract, its fractions and isolated compounds was investigated and a good tyrosinase inhibitory activity was observed for the extract, IC50=85 µg/mL and its polar fractions (ethyl acetate at 60 µg/mL and n-butanol at 75 µg/mL), with gallic acid showing strong anti-tyrosinase activity at IC50 56 µg/mL.

Chemical and biological investigations of Limonium axillare reveal mechanistic evidence for its antidiabetic activity.

Root and bark of Limonium axillare (Forssk) Kuntze are used as antidiabetic remedies in parts of East Africa, but this activity has never been fully investigated. To validate its ethnobotanical use, we compared the chemical and pharmacological profiles of the ethanolic extracts of L. axillare root (REE) and aerial parts (AEE). Administration of REE (500 mg kg-1) reduced streptozotocin-induced hyperglycemia by 44%, restored serum insulin levels, reestablished Glut2 and Glut4 expression and ameliorated pancreatic tissue damage in diabetic rats. In vitro studies revealed a strong radical scavenging effect, α-glucosidase, and α-amylase inhibition activity of REE at IC50 at 25.2, 44.8 and 89.1µg/mL, respectively. HPLC analysis identified ten phenolic compounds in REE with umbelliferone as the major constituents at 10 ± 0.081 mg/g of extract. Additionally, six compounds were isolated from REE including, ß-sitosterol-3-palmitate, ß-sitosterol, myricetin and gallic acids with two new tetrahydrofuran monoterpenes; 2-isopropyl- 3,4,4, trimethyl-tetrahydrofuran (3), and 2-isopropyl-4-methyl-tetrahydrofuran-3,4 dicarboxylic acid (4), the latter was revealed by molecular docking to be a good ligand to glycerol-3-phosphate dehydrogenase a key enzyme in glycolysis.

Anti-estrogenic and anti-aromatase activities of citrus peels major compounds in breast cancer.

Estrogen signaling is crucial for breast cancer initiation and progression. Endocrine-based therapies comprising estrogen receptor (ER) modulators and aromatase inhibitors remain the mainstay of treatment. This study aimed at investigating the antitumor potential of the most potent compounds in citrus peels on breast cancer by exploring their anti-estrogenic and anti-aromatase activities. The ethanolic extract of different varieties of citrus peels along with eight isolated flavonoids were screened against estrogen-dependent breast cancer cell lines besides normal cells for evaluating their safety profile. Naringenin, naringin and quercetin demonstrated the lowest IC50s and were therefore selected for further assays. In silico molecular modeling against ER and aromatase was performed for the three compounds. In vivo estrogenic and anti-estrogenic assays confirmed an anti-estrogenic activity for the isolates. Moreover, naringenin, naringin and quercetin demonstrated in vitro inhibitory potential against aromatase enzyme along with anticancer potential in vivo, as evidenced by decreased tumor volumes. Reduction in aromatase levels in solid tumors was also observed in treated groups. Overall, this study suggests an antitumor potential for naringenin, naringin and quercetin isolated from citrus peels in breast cancer via possible modulation of estrogen signaling and aromatase inhibition suggesting their use in pre- and post-menopausal breast cancer patients, respectively.

Phytochemical profile, enzyme inhibition activity and molecular docking analysis of Feijoa sellowiana O. Berg.

Feijoa sellowiana leaves and fruits have been investigated as a source of diverse bioactive metabolites. Extract and eight metabolites isolated from F. sellowiana leaves were evaluated for their enzymatic inhibitory activity against α-glucosidase, amylase, tyrosinase, acetylcholinestrerase and butyrylcholinesterase both in vitro and in silico. Feijoa leaves' extract showed strong antioxidant activity and variable levels of inhibitions against target enzymes with a strong anti-tyrosinase activity (115.85 mg Kojic acid equivalent/g). Additionally, α-tocopherol emerged as a potent inhibitor of AChE and BChE (5.40 & 10.38 mmol galantamine equivalent/g, respectively). Which was further investigated through molecular docking and found to develop key enzymatic interactions in AChE and BChE active sites. Also, primetin showed good anti BChE (11.70 mmol galantamine equivalent/g) and anti-tyrosinase inhibition (90.06 mmol Kojic acid equivalent/g) which was also investigated by molecular docking studies. Highlights Isolation of eight bioactive constituents from Feijoa sellowiana leaves. In vitro assays using different enzymatic drug targets were investigated. In silico study was performed to define compound interactions with target proteins. Feijoa leaf is an excellent source of anti-AChE and antityrosinase bioactives.

Evaluation of antidiabetic activity of Morus nigra L. and Bauhinia variegata L. leaves as Egyptian remedies used for the treatment of diabetes.

Morus nigra and Bauhinia variegata are used in the Egyptian folk medicine for their hypoglycemic effects. The standardized ethanolic extracts of both plants caused a significant decrease in fasting blood glucose level at two different doses (250 and 500 mg/kg) in streptozotocin-induced diabetic rats' model. Further, in vitro antioxidant activity and α-glucosidase inhibition assays were conducted as well as the measurement of insulin levels and the biomarkers for both liver and kidney functions in the treated animals. Beneficiary effects of BMLE and BVLE in the treatment of diabetes were found not to be limited to hypoglycemic effect but included preventing liver and kidney tissue damage that are associated with diabetes. A strong inhibition of the α-glucosidase enzyme by both extracts may be a contributing mechanism in the overall anti-diabetic effect that was observed. Further detailed study is needed in the future to explore the mechanism of action of both plants.

A Licorice Roots Extract Induces Apoptosis and Cell Cycle Arrest and Improves Metabolism via Regulating MiRNAs in Liver Cancer Cells.

Targeting altered metabolism in cancer provides a promising preventive and therapeutic approach. Natural products interplay between gene expression and metabolism either by targeting altered metabolic enzymes and/or affecting the regulating miRNAs. Licorice is a widely known product used as flavoring agent. Glycyrrhizin and other metabolites were reported to exert several metabolic benefits. Here, we investigated the effect of licorice roots extract on some metabolic pathways and their regulating miRNAs in hepatocellular carcinoma cells. Our data showed various beneficial effects of licorice roots extract including induction of apoptosis and cell cycle arrest. Second, upregulating tumor suppressor miRNAs; let7a-3p, miR-34c-5p, miR-122-5p, miR-126-3p, miR195-5p, miR-199a-5p, miR-206, and miR-326-5p. Third, inhibiting HIF1α, PI3K and C-Myc and activating AMPK and p53. Fourth, inhibiting enzymes of glycolysis; HK-2, LDH-A and PK-M2; pentose phosphate pathway; G6PD and glutaminolysis; glutaminase. However, such an extract upregulated oncogenic miRNAs; miR-21, miR-221, and miR-222. Although the present data highlights the ability of licorice roots extract to enhance apoptosis and cell cycle arrest and correct altered metabolism, it warns against its unfavorable effects, hence, its use for prevention and therapy should proceed with caution. Further experiments are required to investigate whether a specific bioactive ingredient is responsible for upregulating the oncogenic miRNAs.

Methanolic extracts of a selected Egyptian Vicia faba cultivar mitigate the oxidative/inflammatory burden and afford neuroprotection in a mouse model of Parkinson's disease.

Vicia faba L. is a legume from the family Fabaceae. Ancient Egyptians consumed fava beans thousands of years ago and they are still one of the most popular foods in Egypt. The current study examined the anti-Parkinson effect of 80% methanolic extracts of seeds or sprouts of the fava 'Sakha 3 'cultivar which has been selected based on the total phenol content among three cultivars tested. In addition, the extracts were characterized by reversed-phase high-performance liquid chromatography coupled with diode array detection and quadrupole-time-of-flight-mass spectrometry (RP-HPLC-DAD-QTOF-MS). Three doses (200, 400, and 600 mg/kg) of 80% methanol extracts of seeds or sprouts of the Sakha 3 cultivar were evaluated in rotenone-Parkinsonian mice from behavioral, biochemical, and histopathological aspects. The extract of fava sprouts (600 mg/kg dose) showed the most beneficial effect. It improved motor activity, enhanced striatal dopamine level, and decreased the striatal malondialdehyde, as well as the expression of the inflammatory markers, compared with the rotenone control group and groups receiving lower therapeutic doses of the extracts or L-Dopa. In addition, these findings were supported by a histopathological investigation which indicated that mice treated with the 600-mg/kg dose of the sprout extract showed a low number of degenerated neurons. The application of RP-HPLC-DAD-QTOF-MS and mass/mass spectroscopy enabled the metabolic profiling of the sprouts and seeds of the 'Sakha 3' cultivar. It is obvious that germination increased the amounts of phenolic acids, saponins, and aromatic amino acids, together with a dramatic increase in flavonoids. In conclusion, the 80% methanolic extract of sprouts of the fava "Sakha 3" cultivar may be a promising candidate for treating Parkinsonism if appropriate safety data are available.

Metabolite profiles of Pulicaria crispa and P. incisa in relation to their in-vitro/ in-vivo antioxidant activity and hepatoprotective effect: A comparative mass spectrometry-based metabolomics.

Plants of the genus Pulicaria (family: Asteraceae) are widely used in central Asia and the Middle East for treatment of different human diseases. Ultra performance liquid chromatography coupled to high resolution mass spectrometry (UPLC/MS) was utilized to establish the metabolic profiles of two Pulicaria species: P. crispa and P. incisa. 122 metabolites were identified including flavonoids (37), phenolic acids (22), sesquiterpenes (17), diterpenes (7), and fatty acids (27), with enrichment in methoxylated flavonoids (20), caffeoylquinic acid conjugates (14) xanthane sesquiterpenes (9) and hydroxylated fatty acids (20) in both Pulicaria species. The metabolite profile of P. incisa was characterized by the presence of tri- and tetra-methoxylated flavonoids while xanthane sesquiterpenes were the main chemical markers of P. crispa. Additionally, a novel sesquiterpene acid (dihydropulicaric acid) was annotated in both species based on its MS fragments. Antioxidant activity for P. crispa and P. incisa methanol extracts was assessed in vitro based on DPPH and ABTS assays and further in vivo using chlorpromazine intoxicated rat model. Results revealed that P. incisa extract was more effective in inhibiting both DPPH and ABTS free radicals (IC50 0.36 and 0.52 mg/mL, respectively) than P. crispa (IC50 0.51 and 0.73 mg/mL). In the animal model, antioxidant activity of P. incisa (20 mg/kg/day) was also slightly higher causing a 55 % reduction in MDA levels and 65 % increase in GSH activity compared to untreated animals. Furthermore, both extracts showed a hepatoprotective effect as revealed by improvement in levels of serum biomarkers of liver functions: total bilirubin, alanine transaminase (ALT) and aspartate transaminase (AST) comparable to silymarin at 25 mg/kg/day. These findings were also supported by the preserved integrity of the hepatic tissues of animals receiving either extracts at a dose of 20 mg/kg b.wt. The present study reveals for the potential antioxidant and hepatoprotective effects for Pulicaria in relation to its bioactive metabolites.

Comparative metabolite profiling and antioxidant potentials of seeds and sprouts of three Egyptian cultivars of Vicia faba L.

Fava beans are considered basic food in Egypt and all over the world. They are a source of many bioactive secondary metabolites. Since there are many Egyptian cultivars of fava beans, this study addresses comparative chemical profiling of three Egyptian cultivars of fava beans namely Sakha 3, Nubaria 3, and Giza 843. The study also focuses on the effect of germination on the chemical profile of phenolic compounds and saponins. Sixty-five metabolites were characterized based on their UV spectra, accurate MS, and MS/MS data using RP-HPLC-DAD-ESI-QTOF-MS, including 37 metabolites reported for the first time in fava beans. The observed metabolites are phenolic compounds (flavonoids and phenolic acids), saponins, alkaloids, amino acids, nucleosides, organic acids, sugars, and a jasmonate. In fact, upon quantifying the area of metabolites in their chromatographic profiles, it was remarkable that germination increased the quantities of flavonoids, phenolic acids, and saponins. Moreover, the yield (%) of extracts, TPC, and DPPH increased upon sprouting. The TPC and the relative amounts of flavones, saponins, and amino acids showed a good positive correlation with DPPH. These findings elucidated the first report on the metabolic profiling of these cultivars as well as their antioxidant activities.

Black mulberry fruit extract alleviates streptozotocin-induced diabetic nephropathy in rats: targeting TNF-α inflammatory pathway.

OBJECTIVES: This study was designed to investigate the effect of Morus nigra fruit extract in retarding the progression of diabetic nephropathy in streptozotocin (STZ)-induced diabetic rats. METHODS: Diabetic male Wistar rats were injected with black mulberry fruit extract (BMFE) at doses of 150 and 300 mg/kg body weight. After 4 weeks, microalbuminuria was estimated in addition to serum concentrations of glucose, insulin, creatinine and albumin. KEY FINDINGS: The study revealed a significant amelioration of all the measured parameters in diabetic animals. In addition, MDA, lipid peroxide levels and catalase activity were also improved. The histopathological examination of kidney tissues revealed significant improvement of the pathological changes and glomerular sclerosis in diabetic rats treated with BMFE. Treated rats showed downregulation of TNF-α, vascular cell adhesion molecule-1 (VCAM-1) and fibronectin mRNA expression. CONCLUSION: The ameliorative effect of BMFE on diabetic nephropathy is not only through its potent antioxidant and hypoglycaemic effects but also through its downregulation of TNF-α, VCAM-1 and fibronectin mRNA expression in renal tissues of diabetic-treated rats. Therefore, BMFE as dietary supplement could be a promising agent in improving diabetic nephropathy.

Extract of Bauhinia vahlii Shows Antihyperglycemic Activity, Reverses Oxidative Stress, and Protects against Liver Damage in Streptozotocin-induced Diabetic Rats.

BACKGROUND: Several studies have affirmed the effectiveness of some Bauhinia plants as antihyperglycemic agents. OBJECTIVE: We investigated the possible effect of Bauhinia vahlii leaves extract in reducing hyperglycemia and reversing signs of organ damage associated with diabetes in streptozotocin (STZ) rat model. MATERIALS AND METHODS: Both polar fraction of the B. vahlii leaves (defatted ethanolic extract [DEE]) and nonpolar fraction (n-hexane extract) were evaluated in vitro for α-glucosidase inhibition and 2,2-diphenyl-1-picrylhydrazyl radical scavenging potential. DEE was selected for further in vivo studies and was administered at two doses, i.e., 150 or 300 mg/kg to STZ-diabetic rats for 4 weeks. RESULTS: Only DEE exhibited in vitro antioxidant and antihyperglycemic activities and its oral administration at both dose levels resulted in significant reduction in fasting blood glucose and glycated hemoglobin. Furthermore, signs of oxidative stress as indicated by hepatic reduced glutathione, nitric oxide, and malondialdehyde levels were completely reversed. In addition, histopathological examination and measurement of serum aspartate transaminase and alanine transaminase levels showed that DEE protected the liver from signs of liver pathogenesis when compared to diabetic untreated animals and those treated with metformin. Phytochemical analysis of DEE showed high flavonoids content with quercitrin as the major constituent along with other quercetin glycosides. CONCLUSION: This study strongly highlights the possible beneficial effect of B. vahlii leaves extract in relieving hyperglycemia and liver damage in STZ-diabetic rats and recommends further investigation of the value of quercetin derivatives in controlling diabetes and ameliorating liver damage associated with it. SUMMARY: The polar fraction of the Bauhinia vahlii leaves (defatted ethanolic extract [DEE]) exhibited both in vitro antioxidant activity in 2,2-diphenyl-1-picrylhydrazyl scavenging assay and strong α-glucosidase inhibition while the nonpolar fraction (n-hexane extract) failed to show any activity in both assays. DEE was further investigated in streptozotocin-induced diabetic rat model where oral administration of DEE at 2 doses (150 and 300 mg/kg) for 4 weeks resulted in significant reduction in fasting blood glucose and glycated hemoglobin and reversal of oxidative stress signs as indicated by measurement of hepatic reduced glutathione, nitric oxide, and malondialdehyde levels. In addition, histopathological examination and measurement of serum aspartate transaminase and alanine transaminase levels showed that DEE protected the liver from signs of pathogenesis observed in diabetic untreated rats. Phytochemical analysis of DEE showed high flavonoid content with quercitrin as the major constituent (62.9 ± 0.18 mg/mg). Abbreviations used: ALT: Alanine transaminase, AST: Aspartate transaminase, DEE: Defatted ethanol extract, DPPH: 2,2-diphenyl-1-picrylhydrazyl, FBG: Fasting blood glucose, GAE: Gallic acid equivalent, GSH: Reduced glutathione, Hb1Ac: Glycated hemoglobin, HE: Hexane extract MDA: Malondialdehyde, QE: Quercetin equivalent, STZ: Streptozotocin, TAC: Total antioxidant capacity.

Antiprotozoal activity of major constituents from the bioactive fraction of Verbesina encelioides.

The bioactive petroleum ether fraction of Verbesina encelioides, previously studied by the authors, was chosen for the isolation of antiprotozoal metabolites. Pseudotaraxasterol-3ß-acetate (1), benzyl 2,6-dimethoxy benzoate (2), 16ß-hydroxy-pseudotaraxasterol-3ß-palmitate (3) and pseudotaraxasterol (4), in addition to ß-sitosterol glucoside (5) and ß-sitosterol galactoside (6) were isolated and identified based on one-dimensional and two-dimensional spectral analysis. This is the first report describing (3) and (6) in genus Verbesina. The isolated compounds were tested in vitro against Plasmodium falciparum, Trypanosoma brucei, Trypanosoma cruzi and Leishmania infantum. Cytotoxicity was evaluated on MRC-5 cells. Compound 1 showed moderate to weak activity against L. infantum T. brucei and P. falciparum and was inactive against T. cruzi. Compound 3 showed moderate activity against L. infantum, compound 4 revealed weak activity against T. cruzi, while 5 and 6 were inactive against all tested protozoa. All compounds were non-cytotoxic. The isolated constituents showed less antiprotozoal activity than the crude fraction.