Mona H. Hetta

Improvement of lipid profile and antioxidant of hypercholesterolemic albino rats by polysaccharides extracted from the green alga Ulva lactuca Linnaeus.

Sulfated polysaccharides from Ulva lactuca were extracted in hot water and precipitated by ethanol then orally gavaged to rats fed on a hypercholesterolemic diet for 21 days to evaluate the antihypercholesterolemic and antioxidant actions. Atorvastatine Ca (Lipitor) was used as a reference drug. The intragastric administration of U. lactuca extract to hypercholesterolemic rats caused significant decrease of serum total lipids, triglycerides, total cholesterol, LDL-cholesterol and vLDL-cholesterol levels. Whereas, HDL-cholesterol concentration was markedly increased by 180%. Aqueous extract showed a significant ameliorative action on elevated atherogenic index, creatine kinase and lactate dehydrogenase activities of hypercholesterolemic group. Furthermore, serum activities of transaminases and alkaline phosphatase were also improved. High fat diet intake caused a highly significantly elevated serum urea, creatinine concentration. These effects were reversed by oral administration of U. lactuca extract. Sulfates polysaccharides extract of U. lactuca ameliorate hepatic enzymatic (catalase, glutathione peroxidase and superoxide dismutase), non-enzymatic (reduced glutathione & total thiol) antioxidant defenses and thiobarbituric acid reactive substances. In conclusion, the tested U. lactuca polysaccharides extract has potent hypocholesterolemic and antioxidant effects in experimentally-induced hypercholesterolemic animal model.

Triterpenoid saponins from Oreopanax guatemalensis

Seven oleanane-type saponins were isolated from the leaves and stems of Oreopanax guatemalensis, together with ten known saponins of lupane and oleanane types. The new saponins were respectively characterized as 3-O-alpha-L-arabinopyranosyl echinocystic acid 28-O-[alpha- L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester, 3-O-beta-D-glucopyranosyl 3beta-hydroxy olean-11,13(18)-dien-28-oic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta- D-glucopyranosyl]ester, 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl]3beta-hydroxy olean-11,13(18)-dien-28-oic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester, 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl]3beta, 23 dihydroxy olean-18-en-28-oic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-6-O-acetyl glucopyranosyl-(1-->6)-beta-D-glucopyranosyl]ester, 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl] hederagenin 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-[beta-D-xylopyranosyl-(1-->2 )-]beta-D-glucopyranosyl]ester, 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl]hederagenin 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-[beta-D-glucopyranosyl-(1-->2)-]beta-D-glucopyranosyl] ester and 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl] hederagenin 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-[alpha-L-arabinofuranosyl-(1-->2)]-beta-D-glucopyranosyl] ester. The structures were determined by spectral analyses. The NMR assignments were made by means of HOHAHA, 1H-1H COSY, HMQC, HMBC spectra and NOE difference studies.

Estrogenic activity of Citrus medica L. leaves growing in Egypt

The estrogenic activity of petroleum ether extract of Citrus medica L. leaves as well as the chemical constituents responsible for the biological activity was studied. The petroleum ether extract proved to retain high estrogenic activity in immature female rats. The extract was saponified and its components (saponifiable part 23% and the unsaponifiable matter 77%) were identified using GC/MS technique. The extract proved to be safe (LD50< 2g/kg.bw). Oral administration of petroleum ether extract of C. medica in ovariectomized immature female Wistar rats for 7 days in a dose of 400 mg/kg resulted in significant increase in the uterine weight (g) (1.7±0.11) when compared with ovariectomized control rats (1.3±0.07). GC/MS analysis of both saponifiable and unsaponifiable matters revealed the presence of thirty three components (28 hydrocarbons and 5 sterols) in the unsaponifiable fraction, the major hydrocarbon was nHeneicosane (16.7%) while the major sterol was β-sitosterol (4.03%) and 15 components in the saponifiable matter its major component was hexadecanoic acid (19.93%). As a conclusion petroleum ether extract of Citrus medica L. leaves possess a significant estrogenic activity.

Genetic Profiling, Chemical Characterization and Biological Evaluation of Two Conyza Species Growing in Egypt

The present report is a comparative investigation of two Conyza species growing wild in Egypt namely, Conyza dioscoridis (L.) Desf. and Conyza bonariensis (L.) Cronquist. It comprises a genetic and chemical characterization of the plants, as well as an evaluation of their biological activities. The DNA fingerprints of the two species were established based on a polymerase chain reaction (PCR) procedure using ten decamer primers. Further characterization of the plants was performed via determination of pharmacopœial constants, phytochemical screening and estimation of phenolic content (total phenolics, tannins and flavonoids). The ethanol (70%) extracts of C. dioscoridis (EECD) and C. bonariensis (EECB) were subjected to acute toxicity study to determine their LD50; the anti-inflammatory, antimicrobial and cytotoxic activities were then evaluated. Screening for potential cytotoxicity was carried out both by Brine Shrimp Lethality Test and Sulphorodamine-B assay on three human cell lines viz., breast carcinoma (MCF7), colorectal carcinoma (HCT116) and cervical carcinoma (HELA) cell lines. The DNA profiling revealed a similarity index of 88.89% between the investigated species. The variability observed among the pharmacopoeial constants constitute a valuable differential criterion; the total ash, acid insoluble ash, water soluble ash and crude fiber values obtained for C. bonariensis exceeded (17, 5, 10 and 3.5%, respectively) those for C. dioscoridis; meanwhile, the moisture content was higher (10%) in the latter. The phytochemical screening of EECD and EECB revealed the presence of flavonoids, steroids, terpenoids and tannins in both species. Estimation of phenolic contents (total phenolics, tannins and flavonoids expressed as gallic acid, tannic acid and rutin equivalents, respectively) showed that EECD contains higher amounts of all these constituents when compared to EECB (1.17 vs. 0.96 mg/g, total phenolics; 2.43 vs. 1.83 mg/g, tannins and 0.62 vs. 0.29 mg/g, flavonoids). EECD and EECB were found to be safe (LD50 upto 0.5g/kg). Throughout evaluation of the antimicrobial activity against a set of microbial strains and potential cytotoxicity against MCF7, EECD appeared more efficient (MIC: 200-400 µg/ml and IC50: 2.97 μg/ml, respectively); meanwhile, the effect of EECB was more significant on HCT116 and HELA (IC50: 21 and 5.4 μg/ml, respectively). Results of in-vivo assessment of the anti-inflammatory activity showed that the inhibitory effect of EECD was more prominent than that of EECB (74.20% vs. 59.0%). However, the effect of the extracts was inversed in the Brine Shrimp Lethality test (30% vs. 40% lethality, respectively).

Two new dihydroamentoflavone glycosides from Cycas revoluta

Phytochemical investigation of the ethyl acetate extract of Cycas revoluta Thunb. leaflets afforded five compounds including two new dihydroamentoflavone glucosides, (2S)-I-(2,3)-dihydro-I-7-O-β-d-glucopyranosylamentoflavone (1) and (2S)-I-(2,3)-dihydro-I-7,II-7-di-O-β-d-glucopyranosylamentoflavone (2), in addition to the known compounds prunin (3), vitexin-2″-rhamnoside (4) and protocatechuic acid (5). Compounds (3) and (4) being reported for the first time in this plant. The structures of these compounds were established by the detailed analysis of their spectroscopic data, mainly 1D NMR, 2D NMR, CD and HR-MSD-TOF. The ethyl acetate extract showed weak cytotoxicity against HepG2 (IC50 = 207.6 μg/mL) and RAW 264.2 cells (IC50 = 160.8 μg/mL). Compound 4 showed significant activity towards Leishmania donavani (IC50 = 13.8 μM, IC90 = 34.6 μM). The isolated compounds showed weak antimicrobial activity (IC50>10 μg/mL).

The fatty acid-rich fraction of Eruca sativa (rocket salad) leaf extract exerts antidiabetic effects in cultured skeletal muscle, adipocytes and liver cells

Abstract Context: Eruca sativa Mill. (Brassicaceae), commonly known as rocket salad, is a popular leafy-green vegetable with many health benefits. Objective: To evaluate the antidiabetic activities of this plant in major insulin-responsive tissues. Materials and methods: Five E. sativa leaf extracts of varying polarity were prepared (aqueous extract, 70% and 95% ethanol extracts, the n-hexane-soluble fraction of the 95% ethanol extract (ES3) and the defatted 95% ethanol extract). Eruca sativa extracts were investigated through a variety of cell-based in vitro bioassays for antidiabetic activities in C2C12 skeletal muscle cells, H4IIE hepatocytes and 3T3-L1 adipocytes. Guided by the results of these bioassays, ES3 was fractionated into the saponifiable (SM) and the unspaonifiable (USM) fractions. Glucose uptake was measured using [3H]-deoxy-glucose, while the effects on hepatic glucose-6-phosphatase (G6Pase) and adipogenesis were assessed using Wako AutoKit Glucose and AdipoRed assays, respectively. Results: ES3 and its SM fraction significantly stimulated glucose uptake with EC50 values of 8.0 and 5.8 μg/mL, respectively. Both extracts significantly inhibited G6Pase activity (IC50 values of 4.8 and 9.3 μg/mL, respectively). Moreover, ES3 and SM showed significant adipogenic activities with EC50 of 4.3 and 6.1 μg/mL, respectively. Fatty acid content of SM was identified by GC-MS. trans-Vaccenic and palmitoleic acids were the major unsaturated fatty acids, while palmitic and azelaic acids were the main saturated fatty acids. Discussion and conclusion: These findings indicate that ES3 and its fatty acid-rich fraction exhibit antidiabetic activities in insulin-responsive cell lines and may hence prove useful for the treatment of type 2 diabetes.

Effect of 6-gingerol on AMPK- NF-κB axis in high fat diet fed rats

OBJECTIVES Adenosine monophosphate (AMP)-activated protein kinase (AMPK) plays a central role in metabolic homeostasis and regulation of inflammatory responses through attenuation of nuclear factor kappa-B (NF-κB), Thus AMPK may be a promising pharmacologic target for the treatment of various chronic inflammatory diseases. We examined the effect of 6-gingerol, an active ingredient of ginger on AMPK-NF-κB pathway in high fat diet (HFD) rats in comparison to fish oil. METHODS Protein levels of AMPK-α1 and phosphorylated AMPK-α1 were measured by western blot while Sirtuin 6 (Sirt-6), resistin and P65 were estimated by RT-PCR, TNF-α was determined by ELISA, FFAs were estimated chemically as well as the enzymatic determination of the metabolic parameters. RESULTS 6-Gingerol substantially enhanced phosphorylated AMPK-α1 more than fish oil and reduced the P65 via upregulation of Sirt-6 and downregulation of resistin, and resulted in attenuation of the inflammatory molecules P65, FFAs and TNF-α more than fish oil treated groups but in an insignificant statistical manner, those effects were accompanied by a substantial hypoglycemic effect. CONCLUSION Gingerol treatment effectively modulated the state of inflammatory privilege in HFD group and the metabolic disorders via targeting the AMPK-NF-κB pathway, through an increment in the SIRT-6 and substantial decrement in resistin levels.

Comparative DNA profiling, phytochemical investigation, and biological evaluation of two Ficus species growing in Egypt.

Aim and Background: A comparison between two Ficus species, cultivated in Egypt, was carried out in this study. Their DNA analysis revealed that they are not closely related. Materials and Methods: The pharmacopoeial constants of the leaves showed higher total ash and acid insoluble ash in F. lyrata than in F. platypoda. The other parameters were close in both species. Preliminary phytochemical screening revealed the presence of carbohydrate and/or glycosides, tannins, flavonoids, sterols, and triterpenes in their leaves and was detected in traces in their stems. Results: Saponification of n-hexane extract of the leaves yielded 46% and 74.8% for the unsaponifiable matters and 20% and 15% for the fatty acids for F. platypoda and F. lyrata, respectively. n-Docosane (21.69%) and n-heptacosane (33.77%) were the major hydrocarbons in F. platypoda and F. lyrata, respectively. b-Sitosterol was the main sterol, palmitic (22.07%) and carboceric (35.72%) acids were the major identified saturated fatty acids in both species, while linoleic acid was the main unsaturated fatty acid (18.66% and 16.7%) in both species, respectively. The acute toxicity study revealed that the two species were safe up to 2 g/kg. The antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and pyrogallol as the standard was more significant for F. platypoda (232.6 μg/ml) than for F. lyrata, (790.9 μg/ml). The oral antihyperglycemic activity in diabetic rats using alloxan revealed that the 80% ethanolic extract of the leaves of F. platypoda was more active than that of the leaves of F. lyrata in decreasing the blood glucose level at 200 mg/kg/day (107.9 ± 5.817, 127.2 ± 4.359) and 400 mg/kg/day (64.11 ± 4.358, 127.7 ± 6.889), respectively, when compared with the diabetic control gliclazide (172.3 ± 2.089). Conclusion: The results of this study provide evidence that the two Ficus species have antioxidant and antihyperglycemic activity, in the order F. platypoda and then F. lyrata

Acylated Iridoids and Rhamnopyranoses from Premna odorata (Lamiaceae) as Novel Mesenchymal–Epithelial Transition Factor Receptor Inhibitors for the Control of Breast Cancer

Phytochemical investigation of Premna odorata Blanco, Lamiaceae, leaves afforded three new acylated iridoid glycosides 1–3 and two new acylated rhamnopyranoses 9 and 10, in addition to ten known compounds. The structures of the new compounds were confirmed using extensive 1D and 2D NMR analysis. Molecular modeling study suggested the potential of the acylated rhamnopyranoses to bind at the c‐Met kinase domain. Cell‐free Z′‐LYTE™ assay testing revealed the good c‐Met phosphorylation inhibitory activity of 9, followed by 8, and 10, with IC50 values of 2.5, 6.9, and 12.7 μM, respectively. The (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) cell proliferation assay testing against the human c‐Met expressing highly invasive MDA‐MB‐231 suggested compound 9 as the most active with IC50 value of 13.3 μM. Testing of compound 9 against multiple phenotypic breast cancer cell lines including MCF‐7, BT‐474 cells, and MDA‐MB‐468 proved enhanced activity against the highly c‐Met expressing triple‐negative breast cancer cell lines. Acylated rhamnopyranoses are potential novel c‐Met inhibitors appropriate for future optimizations to control c‐Met‐dependent breast malignancies. Copyright

Premna odorata Volatile Oil as a New Mycobacterium tuberculosis Growth Inhibitor for the Control of Tuberculosis Disease

Aims: This study aimed to identify and compare Premna odorata Blanco volatile oil (VO) for the first time; isolated from different plant organs (leaves, young stems, and flowers) with evaluating the oil antituberculosis (anti-TB) activity. Study Design: Experimental design was carried out by using hydrodistillation method, GC/MS analysis and MeDipro Mycobacterium tuberculosis (MTB) Antigen ELISA Technique (MMA-ELISA) accompanied by polymerase chain reaction (PCR) analysis. Place and Duration of Study: This study was carried out at Faculty of Pharmacy, Beni-Suef University, between May to July 2017. Methodology: P. odorata VO was identified using GC/MS analysis, the oil anti-TB activity was evaluated using in vitro and in vivo MMA-ELISA accompanied by PCR analysis. Original Research Article Elmaidomy et al.; EJMP, 21(4): 1-11, 2017; Article no.EJMP.38375 2 Results: GC/MS analysis revealed that P. odorata VO consisted of monoterpenes, sesquiterpenes, diterpenes and higher alkanes; where monoterpenes and sesquiterpenes were represented the major oils fractions. Trans-caryophyllene (29.403% & 14.638%) and β-phellandrene (22.390% & 11.701%) were the major compounds in the leaves and young stems oils, respectively. While αpinene (38.160%) was a characteristic component of the flowers oil. MMA-ELISA showed that a dose of 100 μl/ml in vitro and 300 μl/ml in vivo; the leaves, young stems, and the flowers oils separately had significant anti-TB activities with measured values > 1.5 μg/ml MTB antigen; while the three organs oils in combination 1:1:1 increased the potency of the oils against MTB with measured values < 1.5 μg/ml MTB antigen with PCR negative analysis. Conclusion: P. odorata VO exhibited Anti-TB activity which Anti-TB could be related to the presence of cyclic terpenes (major) and acyclic oxygenated terpenes (minor) compounds.