Transplantation of artificial gelatin-co-bletillastriata gelatin/Salvia miltiorrhiza Corium promotes dermal repair in rats

Purpose: To evaluate the growth of the composite corium (constructed with fibroblast cells and gelatinco- Bletillastriata gelatin/Salvia miltiorrhiza materials) on rats. Methods: The composite artificial corium was constructed by culturing fibroblast cells in gelatin-co- Bletillastriata gelatin/Salvia miltiorrhiza materials. Full-thickness area of skin was excised from the mice and subsequently, the composite corium was transplanted on the wound. Thereafter, the growth difference of the composite artificial corium and natural corium were compared. In addition, real-time fluorogenic reverse transcription polymerase chain reaction (qRT-PCR) and western blot were performed to determine vascular endothelial growth factor (VEGF) expression at gene and protein levels. Results: The composite artificial corium showed significant repair promoting effect on the skin, and the structure of the repaired skin was similar to that of natural corium. Interestingly, PCR and western blot results showed that the expressions of VEGF were higher in composite artificial corium than in natural corium on days 3 and 7 post-transplantation. Conclusion: The composite artificial corium has some clinical prospects for use in the treatment of wounds on large areas of skin.

Antilipolytic and hypotriglyceridemic effects of dietary Salvia triloba Lf (Lamiaceae) in experimental rats

Purpose: Pancreatic triacylglycerol lipase (PL) is a noteworthy pharmacological target for the management of dyslipidemia, and diabetes and obesity. This study was aimed to evaluate the modulatory effects of Salvia triloba L.f. (Lamiaceae) leaves methanol extract (ME) on a high fat diet (HFD)-induced hypertriglyceridemia in rats, with complementary in vitro evaluation of sage PL-inhibitory potential. Methods: Pre-induction of HFD hypertriglyceridemia sage leaves ME (750 mg/kg) was orally supplemented (via gastric intubation) to overnight fasting rats (n = 5). Potential plant modulation of PL was also quantified in vitro by a colorimetric assay (n = 3). For comparison, the effect of Orlistat was similarly evaluated as reference standard. Results: Compared to Orlistat, supplementation of S. triloba at a dose of 750 mg/kg b.wt significantly reversed the HFD-induced postprandial hypertriglyceridemia in experimental overnight fasting rats (p < 0.001 vs. HFD rats). Dietary sage caused 66.4 % reduction in plasma triglycerides. Compared to Orlistat which exerted antilipolytic activity, with half-maximal inhibitory concentration (IC50) of 0.114 ± 0.004 μg/mL), sage inhibited PL activity in vitro in a dose-dependent manner IC50 of 100.80 ± 9.07 μg/mL) Conclusion: Sage has dual hypotriglyceridemic and antilipolytic properties which indicate that it can potentially be used to suppress body weight gain.

Chemical profile and antifungal potential of essential oils from leaves and flowers of Salvia algeriensis (Desf.): A comparative study

Salvia is a plant genus widely used in folk medicine in the Mediterranean area since antiquity. A large number of Salvia essential oils have been reported against diverse microorganisms. In the current study, chemical composition of essential oils from leaves and flowers of Salvia algeriensis (Desf.) was determined using gas chromatography-electron impact mass spectrometry (GC-EIMS) as well as their antifungal activity against phytopathogenic fungi Alternaria solani and Fusarium oxysporum exploring disk method. The GC-EIMS analysis identified 59 compounds (84.8%) in the essential oil obtained from leaves of S. algeriensis. Its major constituents were benzaldehyde (9.7%), eugenol (8.7%) and phenylethyl alcohol (8.4%). In flowers oil, 34 compounds (92.8%) were detected. The main ones were viridiflorol (71.1%) and globulol (8.6%). The essential oil obtained from leaves exhibited the highest antifungal activity, where the effective dose inhibiting 50% of mycelial fungal (ED50) against A. solani was 0.90 μL mL-1 with minimum inhibitory concentration (MIC) equal to 2 μL mL-1, whereas the ED50 and MIC in F. oxysporum culture was 1.84 μL mL-1 and 3 μL mL-1 respectively. The mycelial inhibition by flowers oil varies from 1.77 μL mL-1 (ED50) with A. solani culture (MIC 6.5 μL mL-1) to the lowest effect recorded (ED50 3.00 μL mL-1 and MIC 9.33 μL mL-1) against F. oxysporum. To our best knowledge, this is the first report on S. algeriensis, their leaves oil can constitute an alternative biocontrol against phytopathogenic fungi commonly controlled by chemical fungicides.

Behavioural studies on the ethanol leaf extract of Grewia carpinifolia in Wistar rats.

Background: Grewia carpinifolia is a plant commonly used in the tropics to manage various central nervous system (CNS) disorders. However, despite its widespread use no scientific work has been reported to validate these claims. Objectives: To evaluate the activity of G. carpinifolia as it affects behaviour using animal model. Methods: Twenty five adult Wistar rats were randomly divided into five groups (A-E). Group A served as control (given only distilled water), Groups B,C, D and E were administered with single oral dose of ethanol extract of G. carpinifolia leaf at 100, 200, 400 and 800 mg/kg body weight respectively for twenty eight days consecutively. Subsequently, open field test, negative geotaxis and hanging wire test were performed. Body and brain weights were measured and histological examination of the brain was also performed. Results: At the tested doses, the extract significantly increased the time spent on the hanging wire and decreased locomotor activity at 800 mg/kg. No significant difference was observed in body and brain weights of extract treated groups when compared with the control. No visible histological lesion was also observed. Conclusion: The plant extract may improve muscular strength at tested doses and possess CNS depressant activity at 800 mg/ kg.

Antioxidant and antimicrobial activities of solvent fractions of Vernonia cinerea (L.) Less leaf extract.

Background: Vernonia cinerea (L.) Less is used in folk medicine as a remedy for various diseases. Objectives: The present study reports antioxidant and antimicrobial activities of solvent fractions of Vernonia cinerea. Methods: The antioxidant properties of solvent fractions of V. cinerea were evaluated by determining radicals scavenging activity, total flavonoid and phenolic contents measured with the 2,2-diphenyl-1-picryl hydrazyl (DPPH) test, the aluminum chloride and the Folin-ciocalteau methods, respectively. Antimicrobial activities were tested against human pathogenic microorganisms using agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of each active extract were determined. Results: The ethyl acetate fraction having the IC50 value of 6.50 μg/mL demonstrated comparable DPPH radical-scavenging activity with standard antioxidants, gallic acid and quercetin included in the study. All fractions displayed moderate antimicrobial potential against the tested pathogens with the zone of inhibition that ranged from 9.0 to 13.5 mm. The MIC (1.56 mg/mL) and MBC (3.13 mg/mL) indicated highest susceptibility of Candida albicans in all fractions. Conclusion: The results of this study showed that the solvent fractions of V. cinerea possess antioxidant and antimicrobial activities, hence justifying the folkloric use of the plant for the treatment of various ailments in traditional medicine.

Anti-nociceptive and anti-inflammatory potentials of fractions from the extract of Tetracarpidium conophorum leaf in rats and mice

Tetracarpidium conophorum (TC) (Euphorbiaceae) is a perennial woody climbing shrub in low bush forest of some parts of West Africa and used among the natives for relief of ailments accompanying pain and inflammation. In this study, the analgesic and anti-inflammatory effects of the methanolic extract (METC) and fractions (ethyl acetate, F1 and n-hexane, F2) of Tetracarpidium conophorum leaf were evaluated in rat and mice. The analgesic activity was evaluated using acetic acid-induced writhing, formalin-induced paw licking and hot plate test models. Carrageenan-induced paw oedema was used to assess anti-inflammatory activity in rats. The mechanism of action of (TC) was explored by the use of naloxone, a non-selective opioid receptor blocker. The highest analgesic effect was observed in F2 extract at 57.21% inhibition and was further studied on various analgesic and anti-inflammatory models in graded doses. F2 significantly inhibited the late phase of formalin-induced paw licking and prolong hot plate latency at 55±1°C. The n-hexane fraction also significantly inhibited carrageenan-induced paw oedema in rats at 100 and 200mg/kg doses significantly (p< 0.001) and reduced paw licking response by 85.08% compared with control. Naloxone, an opioid receptor antagonist, did not significantly affect the changes observed with n-hexane fraction, thus ruling out the possibility of the involvement of opioid receptors in the analgesic actions of Tetracarpidium conophorum. Phytochemical screening showed that the leaf extracts contain alkaloids, tannins, saponins and cardenolides. The investigations showed that Tetracarpidium conophorum possesses significant anti-nociceptive and anti-inflammatory activities that should be explored.

In vivo antileishmanial activity and chemical profile of polar extract from Selaginella sellowii

The polar hydroethanolic extract from Selaginella sellowii (SSPHE) has been previously proven active on intracellular amastigotes (in vitro test) and now was tested on hamsters infected with Leishmania (Leishmania) amazonensis (in vivo test). SSPHE suppressed a 100% of the parasite load in the infection site and draining lymph nodes at an intralesional dose of 50 mg/kg/day × 5, which was similar to the results observed in hamsters treated with N-methylglucamine antimonate (Sb) (28 mg/Kg/day × 5). When orally administered, SSPHE (50 mg/kg/day × 20) suppressed 99.2% of the parasite load in infected footpads, while Sb suppressed 98.5%. SSPHE also enhanced the release of nitric oxide through the intralesional route in comparison to Sb. The chemical fingerprint of SSPHE by high-performance liquid chromatography with diode-array detection and tandem mass spectrometry showed the presence of biflavonoids and high molecular weight phenylpropanoid glycosides. These compounds may have a synergistic action in vivo. Histopathological study revealed that the intralesional treatment with SSPHE induced an intense inflammatory infiltrate, composed mainly of mononuclear cells. The present findings reinforce the potential of this natural product as a source of future drug candidates for American cutaneous leishmaniasis.

Determination of antioxidant activity, phenolic contents and antiviral potential of methanol extract of Euphorbia spinidens (Euphorbiaceae)

Purpose: This study was aimed to evaluate the antioxidant activity of the methanol extract of Euphorbia spinidens (Euphorbiaceae) and its effect on Herpes simplex virus type-1 (HSV-1) replication. Methods: The methanol extract of aerial parts of E. spinidens collected from Khorasan State in North- Eastern part of Iran was used in this study. Total phenolic, flavonoid contents and the antioxidant activity were evaluated using Folin-Ciocalteu method, aluminum chloride colorimetric method and β- carotene-linoleate model system, respectively. Both the cytotoxic and antiviral effects of the crude extract on Vero cell line were determined by quantifying the viability of Vero cells using 3-(4,5- dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium (MTS) assay. Results: Total phenolic and flavonoid contents of E.spinidens were 70 ± 1 mg of gallic acid equivalent/g of dry extract (mg GAE/g extract) and 49.66 ± 1.00 mg rutin equivalent/g of dry extract (mg RTN/g extract), respectively. Antioxidant activity was 44 ± 1 % compared with the standard, buthylated hydroxytuloene (BHT). The 50 % cytotoxic concentration (CC50) of the extract on Vero cells was 5.072 ± 0.063 mg/ml and its antiviral concentration of 50 % effectiveness (EC50) value was 0.34 ± 0.003 mg/ml. Conclusion: The findings of this study show that the methanol extract of E. spinidens has high content of phenolic and flavonoid compounds with good antioxidant activity. Furthermore, this extract has significant antiviral effect on HSV-1 probably due to the inhibition of viral replication.

Cuminum cyminum Linn (Apiaceae) extract attenuates MPTP-induced oxidative stress and behavioral impairments in mouse model of Parkinson’s disease

Purpose: To evaluate the protective effects of Cuminum cyminum Linn (Apiaceae, CCY) against 1- methyl-4 phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced oxidative stress and behavioral impairments in mouse model of Parkinson’s disease (PD). Methods: MPTP-intoxicated mice model of PD was used for evaluating the effect of CCY extract on behavioral deficits through rota rod, passive avoidance and open field tasks. The effect of CCY extract on oxidative stress levels were assessed by estimating enzyme status, including superoxide dismutase (SOD), catalase (CAT) and lipid peroxidation(LPO) in brain tissues of MPTP-induced mice. Results: MPTP (25 mg/kg, i.p.)-treated mice resulted in a significant (p < 0.001) behavioral deficit in locomotor behavior (from 56.24 ± 1.21 to 27.64 ± 0.94) and cognitive functions (from 298 ± 3.68 s to 207.28 ± 4.12 s) compared with their respective control groups. Administration of CCY extract (100, 200 and 300 mg/kg, p.o.) for three weeks significantly and dose-dependently improved (p < 0.001 at 300 mg/kg) locomotor and cognitive deficits in MPTP-treated mice. CCY treatment also significantly (p < 0.001 at 300 mg/kg) inhibited MPTP-induced decrease in antioxidant enzyme levels (superoxide dismutase and catalase) and lipid peroxides in mice brain tissues. Conclusion: CCY extract exhibits strong protection against MPTP-induced behavioral deficit through enhancement of antioxidant defense mechanisms. Therefore, CCY may be developed as a therapeutic strategy in the treatment of neurodegeneration seen in PD.

Evaluation of a root extract gel from Urtica dioica (Urticaceae) as analgesic and anti-inflammatory therapy in rheumatoid arthritis in mice

Purpose: To develop and characterize an herbal gel prepared from methanol root extract of Urtica dioica (Urticaceae) (Stinging nettle) for the treatment of arthritis in mice. Methods: A methanol root extract from Urtica dioica was prepared, and a gel was then prepared using Carbopol 934. The prepared gel was subjected to various physical tests (color, appearance, pH, texture, viscosity) and in vivo evaluation, including primary skin irritation, analgesic, and anti-inflammatory tests, in arthritic mice and compared with 2 % indomethacin gel, which was used as standard. Results: The prepared herbal gel was of light gray color with a smooth texture. It showed a pH of 7.1 and a viscosity of 21.2 cps. The gel exhibited pseudoplastic rheology, as evidenced by shear thinning with increased shear rate. It was non-irritating to the skin in primary skin irritation test in mice and showed 55.05 % inhibition of paw edema in a carrageenan-induced hind rat paw edema model, comparable to that of the standard gel (53.93 %), after 24 h. The gel showed 58.21 % analgesia, versus 61.19 % analgesia for the indomethacin gel standard in writhing test. Conclusion: The topical gel from methanol root extract of U. dioica may be an efficacious and safe alternative to non-steroidal anti-inflammatory drugs in the treatment of rheumatoid arthritis but this requires further investigations to ascertain its safety and clinical efficacy.

Healing effect of Terminalia chebula Retz extract on second-degree burns in rats

Purpose: To investigate the healing effect of Terminalia chebula Retz Extract (TCRE) on seconddegree burns in rats. Methods: Male Sprague Dawley (SD) rats, weighing 200 – 220 g, were subjected to deep seconddegree skin burns by electrical scald instrument. The animals were divided into three groups as follows: (1) second-degree burns model (control) group, (2) burns model treated with 1 % silver sulfadiazine (SSD) group, and (3) burns model treated with 100 mg·mL-1 TCRE group. On days 3, 7 and 14 following the administration of the drug/extract, the wound area and histopathological changes in rat epidermis were evaluated for the various groups. The minimum inhibitory concentration (MIC) of TCRE on Staphyloccocus aureus, Pseudomonas aeruginosa and Escherichia coli were also assessed separately. Results: On day 14, the mean wound area of TCRE treatment group (0.25 ± 0.06 cm2) was significantly smaller than that of the control rats (2.71 ± 0.20 cm2, p < 0.01). The histological results indicate that the inflammatory cells disappeared and were replaced by new granulation tissue in the group treated with 100 mg·mL-1 TCRE by day 14. Compared with SSD group rats, the inflammatory cells and fibroblast and granulation tissues of burnt rats treated with 100 mg·mL-1 TCRE were same as those of rats that had no burns. The antibacterial results revealed that the MIC of TCRE on Staphyloccocus aureus, Pseudomonas aeruginosa and Escherichia coli was 3.13, 12.5 and 6.25 mg·mL-1, respectively. Conclusion: Terminalia chebula Retz. has potentials to be developed as an effective medicinal herb for the treatment of second-degree burns.

Effect of Arisaema erubescens (Wall) Schott rhizome extract on rheumatoid arthritis

Purpose: To investigate the anti-arthritic activity of the water extract of Rhizoma Arisaematis (WERA) using a collagen II -induced arthritis (CIA) rat model. Methods: CIA was induced in male Sprague-Dawley rats by intradermal injection of bovine collagen II in Complete Freund’s Adjuvant. The rats were treated with daily oral doses of WERA (100, 200, and 400 mg/kg) for 21 consecutive days. Methotrexate (MTX, 3 mg/kg), used as a positive control, was administered orally 2 times/week for 3 weeks. The severity of arthritis was evaluated using indices of paw swelling, arthritic score, body weight, thymus index, and spleen index. In addition, the serum levels of IL-1β, IL-6, IL-10, and TNF-α were measured. Results: All doses of WERA significantly inhibited paw edema (p < 0.01), decreased arthritis scores (p < 0.01) and spleen index (p < 0.05), and alleviated the weight loss associated with CIA in rats. Furthermore, TNF-α, IL-1β, and IL-6 serum levels were significantly decreased (p < 0.05) by all doses of WERA. By contrast, IL-10 serum levels were markedly increased (p < 0.05). Conclusion: WERA exerts therapeutic effects in CIA in rats by decreasing the serum levels of TNF-α, IL-1β, IL-6 and IL-10, suggesting WERA may be an effective candidate drug for treating human rheumatoid arthritis.

Ziziphus mauritiana leaf extract emulsion for skin rejuvenation

Purpose: To formulate stable water in oil (W/O) emulsion containing hydroalcoholic crude extract of Ziziphus mauritiana leaves for skin rejuvenation. Methods: Placebo (base) without any plant extract and formulation with 4 % Ziziphus mauritiana extract were prepared by mixing. Samples of the emulsions were subjected to varying storage conditions, i.e., 8, 25, 40 oC and 40 oC + 75 % relative humidity for a period of 4 weeks to predict their stability. During this period, stability parameters, including liquefaction, phase separation, color, electrical conductivity, centrifugation and pH were monitored at specified time intervals. Skin rejuvenation was evaluated using 13 healthy human volunteers over a period of 8 weeks. During this period, various skin parameters such as erythema, melanin level, moisture content, elasticity and sebum content of the skin were evaluated at specified intervals. Results: Both the active formulation and placebo were stable in terms of liquifaction, phase separation and color at all the storage conditions of temperature and humidity. Active formulation showed statistically significant (p < 0.05) improvement in skin melanin as well as in skin moisture and sebum levels, whereas these properties were reduced or even absent in the placebo formulation (p > 0.05). Both active and placebo formulations changed skin elasticity and erythema significantly (p < 0.05). Conclusion: It is evident from the findings that the leaf extract of Ziziphus mauritiana possesses antiaging properties as well as exert skin lightening, moisturizing and viscoelastic effects on human skin.

Anti-anxiety effect of methanol extract of Pericarpium zanthoxyli using a strychnine-sensitive glycine receptor model

Purpose: To determine if the methanol extract of Pericarpium zanthoxyli exerts anti-anxiety effects and also to explore any probable anti-anxiety mechanism in vivo. Methods: The staircase test, elevated plus maze test, rota-rod treadmill test and convulsions induced by strychnine and picrotoxin on mice were tested to identify potential mechanism of anti-anxiety activity of the plant extract. Results: The plant extract (10 mg/kg, p.o.) significantly reduced rearing numbers in the staircase test while it increased the time spent in the open arms as well as the number of entries to the open arms in the elevated plus maze test, suggesting that it has significant anti-anxiety activity. Furthermore, the extract inhibited strychnine-induced convulsion. However, it had little effect on picrotoxin-induced convulsion, suggesting that its anti-anxiety activity may be linked to strychnine-sensitive glycine receptor and not GABA receptor. Conclusion: These results suggest that the Pericarpium zanthoxyli extract may be beneficial for the control of anxiety.

Chromatographic-mass spectrometric analysis of ethanol extract of Maesa perlaria var formosana

Purpose: This study analyzes the chemical composition of ethanol root extracts of Maesa perlaria var formosana by gas chromatography-mass spectrometry (GC-MS). Methods: The dried root of Maesa perlaria var formosana was extracted with 95 % ethanol for composition analysis under the following optimum GC-MS conditions: 250 °C inlet temperature, 250 °C MSD detector temperature, and GC oven temperature programmed as follows: initial temperature held at 70 °C for 15 min, then increased at a rate of 2.5 °C/min and held at 170 °C for 15 min; then raised at a rate of 2 °C/min and kept at 180 °C for 20 min; then raised at 2 °C/min and kept at 250 °C for 20 min. Finally, it was raised at 3 °C/min and kept at 280 °C for 15 min. Results: A total of 59 chemical compounds were identified, representing 88.82 % of the composition of the ethanol extracts. The three major components, include 2,4-di-tert-butylphenol (16.76 %), stigmasterol (15.86 %) and campesterol (7.33 %). Conclusion: The results show that a total of 59 components were identified in the ethanol extract of Maesa perlaria var. formosana. The major component, 2,4-Di-tert-butylphenol, exhibits various biological activities.