Reinwardtia indica: phytochemical screening and evaluation of wound healing activity of the extracts in experimental model rats.

Reinwardtia indica is traditionally used for wound healing. The main aim of this study was to evaluate the wound healing activity of leaves extracts of R. indica using the excision wound model in rats. The leaves of R. indica were collected from Gondrang, Chitwan, Nepal.  Leaves were shade dried, extracted by double maceration and subjected to phytochemical screening. Then, the fusion method was used for the formulation of ointment and evaluated. Rats (n=24) were divided into four groups with 6 in each. Excision wound model was used, 2 cm diameter (314 mm2), 2 mm depth wound was created. The treatment was given daily topically to all groups and the % mean wound contraction rate was calculated on days 4, 8, 12 and 16.  The result was analyzed statistically using Graph pad prism version 5. Phytochemical test revealed the presence of alkaloid, flavonoid, tannin, phenol, terpenoid, carbohydrate, etc.  All the evaluation parameters showed satisfactory results. The extract of R. indica ointment (2% w/w and 5% w/w) increased the wound contraction rate day by day. The % means wound contraction rate, on day 12, (80% and 88%), and on day 16, (97% and 100%) and statistically significant difference was at p<0.0001. The R. indica extract ointment showed an increased wound contraction rate. So, in further R. indica could be used for commercial production of wound healing ointment.

Neuroprotective effect of Reinwardtia indica against scopolamine induced memory-impairment in rat by attenuating oxidative stress.

Reinwardtia indica belongs to Linaceae family and used as a folk medicine in Asian countries. Traditionally, it has been used in the treatment of paralysis and anti-microbial in wound healing, etc. The current study was undertaken in order to investigate the antioxidant and memory protective effect of the alcoholic (99.90%) (AERI) and hydro-alcoholic (70:30) leaves extract (HAERI) of Reinwardtia indica, against scopolamine-induced memory impairment in animals and also tried to determine the possible mechanism of action. In addition, phytochemical profiling of alcoholic leaves extract was also conducted through gas chromatography-mass spectrometry (GC-MS/MS). Rats were pretreated with AERI, HAERI (dose 250 and 500 mg/kg) and Donepezil (standard drug) along with scopolamine (1 mg/kg) for a period of 14 days followed by different test like elevated plus maze, passive avoidance, and Morris water maze to assess learning and memory ability. Acetylcholine levels, acetylcholinesterase (AChE), antioxidant enzymes (SOD, CAT & GSH), histopathology of the brain and biochemical test were also performed at the end of the treatment period. The scopolamine treatment resulted in learning and memory deficits which were partially and significantly ameliorated by the AERI at higher dose among other doses of extracts. The AERI at higher dose also counteracted the scopolamine-induced decrease in acetylcholine levels, increase in AChE activity, and decrease in antioxidant enzymes activities. No significant changes observed in the biochemical estimation of all dose of extracts. Histology of brain tissue showed the marked cellular changes in only scopolamine treated group while the standard, AERI and HAERI treated group were showing less damage at hippocampus region of the brain. The phytochemicals found after chemical profiling through GC-MS also supported the activity because of the presence of chemicals already reported for the neuroprotective, memory-enhancing and antioxidant activity, etc. The results demonstrated that the ability of the AERI at higher dose among all doses of extracts has more potential to revert the scopolamine-induced learning and memory deficits in rats by attenuating the decreased level of acetylcholine and antioxidant enzymes.

Evaluation of antioxidant and antimicrobial potential of a novel Himalayan plant Reinwardtia indica dumort: Scientifically unexplored.

Reinwardtia indica (Lineceae) is a medicinal plant cultivated in the Himalayan region. It is effectively used in folk medicines for the treatment of various health complications. In the present study, the shade dried leaves and stem were extracted in three different solvents such as ethyl acetate, ethanol, and hydro-alcoholic. The antioxidant efficacy of these extracts was confirmed by using different in vitro assays: DPPH free radical scavenging, superoxide radical scavenging, lipid peroxidation, metal ion chelating capability and reducing power determination. Total phenol content was maximum in hydro alcoholic extract of leaf (540.37 mg per g of gallic acid equivalents) and stem (330.51 mg per g of gallic acid equivalents) while flavonoid content was maximum in ethanolic extract of leaf (305 mg per gram of rutin equivalents) and ethyl acetate extract of stem (170.6 mg per gram of rutin equivalents). The antioxidant activity of these extracts was positively correlated with their total phenol and flavonoid content. Among all tested extracts, ethanolic extract of leaf exhibit maximum zone of inhibition against all tested clinical isolates of bacterial (E. coli 11.00 ±â€¯1.73 mm, P. aeurogenosa 11.67 ±â€¯0.58 mm and S. aureus 10.33 ±â€¯1.53 mm) and fungal (C. albicans 11.33 ±â€¯1.10 mm) pathogens, while ethyl acetate extracts of the leaf and stem showed minimum inhibitory concentration against all tested microorganisms. Thus, R. indica extracts can be used as potent natural antioxidant having antifungal and antibacterial action.

Antioxidant, antimicrobial and cytotoxic potential of silver nanoparticles synthesized using flavonoid rich alcoholic leaves extract of Reinwardtia indica.

The present work discusses the establishment of a green route for the rapid synthesis of silver nanoparticles (AgNPs) using an alcoholic extract of Reinwardtia indica (AERI) leaves which act as a reducing as well as a capping agent. The change in color from yellowish green to dark brown confirmed the synthesis of AgNPs. A characteristic surface plasmon resonance (SPR) band at 436 nm advocated the presence of AgNPs. The synthesis process was optimized using one factor at a time approach where 1.0 mM AgNO3 concentration, 5 mL 0.4% (v/v) of AER inoculum dose and 30 min of sunlight exposure were found to be the optimum conditions. The synthesized AgNPs was characterized by several characterizing techniques such as HR- TEM, SAED, HR-SEM, EDX, XRD, FTIR and AFM analysis. For evaluation and comparison of AgNPs with AERI used human pathogen E. coli, P. aeurogenosa, S. aeurus and C. albicans for antimicrobial, for cytotoxicity study SiHa cell line at concentration of (10, 50, 100, 250 and 500 µg mL-1) and for enzymatic assay superoxide dismutase, catalase, malondialdehyde and glutathione peroxidase method were used. The size of nanoparticle in the range of 3-15 nm was confirmed TEM, spherical shape by SEM and crystal lattice nature by XRD. AFM results revealed the 2 D and 3 D pattern of particle scatter nature on the surface. This protocol as simple, rapid, one step, eco-friendly, nontoxic and AgNPs showed strong antimicrobial activity as well as cytotoxic potential in comparison to AERI.

The diterpenes ovoideal A-G from Tirpitzia ovoidea.

Seven new diterpenes, named ovoideal A (1), B (2), C (3), D (4), E (5), F (6) and G (7), have been isolated along with eleven known diterpenes 8-18 from the petroleum ether soluble fraction of an ethanol extract of the aerial parts of Tirpitzia ovoidea. The structures of the new compounds were elucidated primarily by 1D and 2D NMR spectroscopy, as well as by the HR-ESI-MS spectrometry. All compounds were isolated from the Linaceae family for the first time. The in vitro cytotoxic activity of compounds 1, 3-5, 8-18 was evaluated against the Hela, HepG2 and K562 cell lines. Among them, compounds 3, 9, 11, 12, 13, 14, 15, 17, 18 showed moderate inhibitory activities.

Antifungal rosane diterpenes and other constituents of Hugonia castaneifolia.

The rosane diterpenoids hugorosenone [3beta-hydroxyrosa-1(10),15-dien-2-one], 18-hydroxyhugorosenone and 18-hydroxy-3-deoxyhugorosenone, and 12-hydroxy-13-methylpodocarpa-8,11,13-trien-3-one were isolated as antifungal constituents of H. castaneifolia Engl. root bark, together with the previously reported di-podocarpanoids hugonone A and hugonone B that were weakly active, and 1(10),15-rosadiene-2beta,3beta-diol (hugorosenol), 4alpha-methoxyhimachal-10-en-5beta-ol (hugonianene B) and 2-hydroxyhenpentacont-2-enal, and the known compounds tetracosyl-(E)-ferrulate and caryophyllene oxide, all of which were inactive. Hugorosenone also exhibited activity against Anopheles gambiae mosquito larvae. Structural determination was achieved based on spectroscopic data.

A new cytotoxic and larvicidal himachalenoid, rosanoids and other constituents of Hugonia busseana.

A new cytotoxic himachalene sesquiterpenoid 4alpha-methoxy-5,9-oxahimachal-9-ene (hugonianene A) which exhibited moderate activity against Anopheles gambiae mosquito larvae after 24 h at a concentration of 0.237 mg mL(-1), and at 48 h and 72 h contact time causing complete larval mortality up to a concentration as low as 0.01369 mg mL(-1), was isolated as the major constituent of the cytotoxic root bark extract of Hugonia busseana. Hugonianene A was obtained together with the hitherto unreported rosane diterpenoid 18-hydroxyrosane, the known rosane diterpenoid hugorosediol, an inseparable mixture of 12-methoxy-13-methylpodocarpa-8,11,13-trien-3,7-dione and 12-methoxy-13-methylpodocarpa-1,8,11,13-tetraen-3,7-dione, and the di-podocarpanoid hugonone B that was previously obtained from H. castaneifolia.

Lignans and diterpenes isolated from Tirpitzia ovoidea and their biological activities.

A new lignan, tirpitzin A (17) together with 20 known compounds (1-16, and 18-21) were isolated from the ethyl acetate soluble fraction of ethanol extract of the aerial parts of Tirpitzia ovoidea. The structure of new compound was elucidated by means of spectroscopic analysis. Of the known compounds, 7-21 were isolated from Linaceae family for the first time. The pharmacological activity of the crude extracts was tested using a mouse inflammation model induced by dimethyl benzene. The results demonstrated that the ethyl acetate soluble fraction had anti-inflammatory activity. Moreover, the cytotoxic and anti-inflammatory activities of some compounds were studied. The new compound 17 showed moderate cytotoxic effect against BxPC-3 cell line (IC50 = 19.51µmol·L-1) and Compound 10 showed significant cytotoxicity against HepG2, HL-60, U87 and BxPC-3 cell lines with IC50 values in the range 4.2-8.3µmol·L-1. Additionally, Compounds 2, 10, 11, and 13 exhibited potent inhibitory effects on LPS-induced nitric oxide production in RAW 264.7 macrophages at the concentration of 50µmol·L-1.