Ajuga Bracteosa Wall Research Articles

Characterization and comparative evaluation of wound healing potential of Ajugarin I and Ajuga bracteosa Wall. ex Benth.

Ajuga bracteosa (family: Lamiaceae), commonly known as kauri booti, is an important ethnomedicinal plant. The current research was conducted to appraise and compare the in vitro antioxidant and antibacterial profiles as well as in vivo wound healing potentials of Ajugarin I and A. bracteosa extract. Ajugarin I and polyphenols in A. bracteosa were enumerated by reversed-phase high-performance liquid chromatography analysis that confirmed significant amounts of Ajugarin I (2.2 ± 0.02μg/mg DW) and other phenolic compounds (14 out of 17 standards). A. bracteosa (374.4 ± 0.20µg AAE/mg of DW, 201.9 ± 0.20µg AAE/mg of DW, 87 ± 0.30%) showed a higher antioxidant profile compared to Ajugarin I (221.8 ± 0.50µg AAE/mg of DW, 51.8 ± 0.40µg AAE/mg of DW, 27.65 ± 0.80%) with 1.86-, 3.89-, and 3.15-fold greater activity in ferric reducing antioxidant power, total antioxidant capacity, and free radical scavenging assays, respectively. Likewise, A. bracteosa showed antibacterial activity against 3/5 strains (MIC 25-200μg/ml) than Ajugarin I (2/5 strains; MIC 50-200μg/ml). Hemolytic (<2% hemolysis) and dermal toxicity tests rendered both samples non-toxic. Additionally, A. bracteosa (100 ± 2.34% at day 12; 9.33 ± 0.47days) demonstrated 1.11- and 1.24-fold higher percent wound contraction and epithelization time, respectively, than Ajugarin I (95.6 ± 1.52% at day 12; 11.6 ± 0.47days) as assessed by an excision wound model in mice. Histopathological examination further reinforced the better wound healing potential of A. bracteosa with good epithelization, collagen synthesis, fibroblast proliferation, and revascularization. Briefly, we endorse the significant comparative antioxidant, antibacterial, and wound healing activities of A. bracteosa and Ajugarin I and present these as prospective candidates for wound healing drugs.

Nanoscale zeolitic imidazolate framework–8 encapsulates crude extract of Ajuga bracteosa wall ex. Benth and enhanced their antibacterial efficiency

Certain disease-causing bacterial strains invade the body; they multiply and cause disease. The development of nanoparticles and self-assembled nanomaterial-based therapeutics has developed into a flourishing section in nanomedicine. Zeolitic imidazole framework-8 (ZIF-8) comprises zinc ions and 2-methylimidazole showing good biocompatibility, known for their large porosity with high surface area, tunable shapes, large pore sizes, and controllable surface functionality. We report a ZIF-8 based agent with excellent antibacterial effectiveness against gram-positive (Bacillus subtilis) and gram-negative (Pseudomonas aeruginosa) bacteria by agar diffusion method. We are the first to encapsulate Ajuga bracteosa extract (ABE) into the ZIF-8 framework, forming a nanoconjugate (ABE@ZIF-8). The mechanism by which the synthesized ABE@ZIF-8 NPs are inhibiting bacterial growth is likely to be the adsorbance of ABE@ZIF-8 NPs on the bacterial cell membrane because of the electrostatic attraction between positively charged ABE@ZIF-8 NPs and the negatively charged bacterial cell membrane. UV–vis absorption spectra, PXRD, SEM, TEM, BET, TGA, DLS, zeta potential characterized ABE@ZIF-8 NPs. Zone of inhibition (ZOI) and minimum inhibitory concentration (MIC) of ABE@ZIF-8 NPs are studied against the Bacillus subtilis and Pseudomonas aeruginosa compared to gentamicin, ZIF-8, ABE, and methanol. The results reported ABE@ZIF-8 nanoconjugate has better antibacterial efficiency against gram-positive and gram-negative bacteria.

Polyphenol Rich Ajuga bracteosa Transgenic Regenerants Display Better Pharmacological Potential.

Ajuga bracteosa Wall. ex Benth. is an endangered medicinal herb traditionally used against different ailments. The present study aimed to create new insight into the fundamental mechanisms of genetic transformation and the biological activities of this plant. We transformed the A. bracteosa plant with rol genes of Agrobacterium rhizogenes and raised the regenerants from the hairy roots. These transgenic regenerants were screened for in vitro antioxidant activities, a range of in vivo assays, elemental analysis, polyphenol content, and different phytochemicals found through HPLC. Among 18 polyphenolic standards, kaempferol was most abundant in all transgenic lines. Furthermore, transgenic line 3 (ABRL3) showed maximum phenolics and flavonoids content among all tested plant extracts. ABRL3 also demonstrated the highest total antioxidant capacity (8.16 ± 1 μg AAE/mg), total reducing power, (6.60 ± 1.17 μg AAE/mg), DPPH activity (IC50 = 59.5 ± 0.8 μg/mL), hydroxyl ion scavenging (IC50 = 122.5 ± 0.90 μg/mL), and iron-chelating power (IC50 = 154.8 ± 2 μg/mL). Moreover, transformed plant extracts produced significant analgesic, anti-inflammatory, anticoagulant, and antidepressant activities in BALB/c mice models. In conclusion, transgenic regenerants of A. bracteosa pose better antioxidant and pharmacological properties under the effect of rol genes as compared to wild-type plants.

In Vitro Antioxidant Activity, Total Phenolic, Flavonoid and Tannin Contents in the Ajuga Bracteosa Wall. Ex Benth, Grown at Middle Hill Climatic Condition of Western Himalayas

The antioxidant activity of aqueous and alcoholic extracts of different plant parts viz, leaves, flower, stem and root of Ajuga bracteosa was investigated against various in-vitro antioxidant assays. The total phenolic, flavonoid and tannin contents also estimated. The results revealed the significant antioxidant potential and variation in the IC50, EC50 and phytochemical contents among all the plant parts. The aqueous extract of leaves exhibited significantly (P&lt;0.05) highest antioxidant activity on 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acixd), 2,2-diphenyl-1-picryl hydrazyl, and potassium ferricyanide reducing power assay, with IC50 values of 0.2707±0.0008, 0.4409±0.0020, and EC50 value 0.3413±0.0030 mg/mL, respectively, followed by the other parts of the plant. The leaves extract also possess the highest total phenolics, flavonoid and tannin contents among all the parts. Similarly, the aqueous extract is better than the alcoholic extract of different parts as far as phytochemical contents and antioxidant activity concerned. The present study revealed that the aqueous extract of leaves had the highest antioxidant potential, which correlated with the high level of total phenolic fl flavonoid, and tannin contents. Therefore, higher the phytochemical contents, higher will be the antioxidant potential.

Enhanced stress tolerance in transformed Ajuga bracteosa Wall. ex Benth. regenerants by upregulated gene expression of metabolic pathways

The rol oncogenes of Agrobacterium rhizogenes enhance the production of medicinally important compounds in plants and provide a first barrier against the overproduction of reactive oxygen species during biotic and abiotic stress. 'this study was designed to evaluate the expression of genes involved in biosynthetic pathways and their impact on metabolic contents and environmental stress tolerance in regenerated Ajuga bracteosa Wall. ex Benth. After successful transformation, real-time quantitative PCR confirmed the increased expression (1.94-6.59-fold) of HMGR, HDS, FDS, PAL, and TTG1 genes in transgenic lines. Furthermore, GC-MS coupled with principal component analysis revealed diverse concentrations of 97 metabolites in A. bracteosa. Transgenic lines showed greater survival under multiple stresses. This was revealed by significant chlorophyll content (8.13-21 mu moles/m(2)), higher quantum efficiency of PSII (F-v/F-m), and the performance index (PIabs) value. Similarly, catalase and peroxidase enzyme activities were enhanced during extreme drought (300-400 mM mannitol) and salinity (150-200 mM NaCl) conditions, compared to untransformed control. Wild type control plant leaves were completely necrotized by Aspergillus fumigatus (FCBP 66) and Fusarium solani (FCBP 0291), whereas transformed leaves had improved antifungal resistance. In conclusion, our data suggest that rolABC genes have a significant impact on the synthesis of metabolites involved in enhancing multistress tolerance in A. bracteosa.

A review on phytochemical and ethnopharmacological studies of Ajuga Bracteosa Wall. Ex Benth.

Herbal medicines as the major remedy in traditional system of medicine have been used in medical practices since antiquity. The plants of genus Ajuga are evergreen, clump-forming rhizomatous perennial or annual herbaceous flowering species, with Ajuga being one of the 266 genera of the family Lamiaceae. There are at least 301 species of the genus Ajuga with many variations. Ajugabracteosa Wall. ex Benth (A. bracteosa) is an important medicinal plant of Himalaya regions. Medicinal potential is due to presence of of various pharmacologically active compounds such as neo-clerodane diterpenoids, flavonol glycosides, iridoid glycosides, ergosterol-5,8- endoperoxide and phytoecdysones. The aim of this review article was to gather information about A. bracteosa which is currently scattered in form of various publications. This review article tried to attract the attention from people for therapeutic potential of A. bracteosa. The present review comprises upto date information of,traditional uses, botanical aspects, active ingredients and pharmacological activities such as antitumor, antimicrobial, antimalarial, anti-inflammatory, cardiotonic activity, antiarthritic activity, antioxidant activity . A large variety of compounds have so far been isolated from Ajuga bracteosa.&#x0D; Keywords: Ajuga bracteosa, Herbal medicines, pharmacological activities.

Antimicrobial Potential and Determination of Total Phenolic and Flavonoid Content of Aerial Part Extracts of Ajuga bracteosa Wall ex. Benth

Antimicrobial Potential and Determination of Total Phenolic and Flavonoid Content of Aerial Part Extracts of Ajuga bracteosa Wall ex. Benth

Antimycotic activity of ethanolic and aqueous leaf extracts of Ajuga bracteosa Wall. ex Benth. (Lamiale: Lamiaceae) and Iris kashmiriana Baker (Asparagales: Iridaceae) against some vegetable rot fungi

Antifungal activities of different solvent extracts of Ajuga bracteosa Wall. ex Benth. (Lamiale: Lamiaceae) and Iris kashmiriana Baker (Asparagales: Iridaceae) were carried out through agar well diffusion assay at three concentrations (25 uL, 50 uL and 75 uL) against seven rot causing fungi, viz. Penicillium expansum, Aspergillus niger, Mucor plumbeus, Alternaria alternata, Penicillium chrysogenum, Trichothecium roseum and Rhizoctonia solani. All the concentration of plant extracts showed antimycotic activity against tested pathogenic fungi. Antimycotic activity increased with the increased concentrations of plant extracts. However, higher concentrations proved more effective than lower concentrations. It was revealed from the present study that the ethanolic extract of Ajuga bracteosa showed maximum antimycotic activity against Mucor plumbeus and Rhizoctonia solani and least activity against Penicillium chrysogenum. However, the aqueous extract of Ajuga bracteosa showed maximum antifungal activity against Rhizoctonia solani and Penicillium expansum and least activity against Trichothecium roseum. It was further revealed from the present study that the ethanolic extract of Iris kashmiriana showed maximum antimycotic activity against Aspergillus niger and least activity against Rhizoctonia solani. Whereas the aqueous extract of Iris kashmiriana showed maximum antimycotic activity against Penicillium expansum and Rhizoctonia solani and least activity against Aspergillus niger.

Antimutagenic activity of compounds isolated from Ajuga bracteosa Wall ex. Benth against EMS induced mutagenicity in mice.

Ajuga bracteosa Wall ex. Benth. (Lamiaceae) has been reported to possess many biological activities including antibacterial, antifungal, antispasmodic and antioxidant activity but there is no report as such on its mutagenic and/or anti-mutagenic activity. The aim of the present study was to isolate compounds from the methanol extract of the aerial parts of Ajuga bracteosa and determine their anti-mutagenic activity against the mutagen, EMS in animal model mice. The study was undertaken in order to corroborate the traditional use of the plant Ajuga bracteosa. The compounds were isolated from the methanol extract of the aerial parts of Ajuga bracteosa using silica gel column chromatography. Structural elucidation of the isolated compounds was done using spectral data analysis and comparison with literature. High performance liquid chromatography (HPLC) was used for the qualitative and quantitative determination of the isolated compounds in the crude methanol extract. The isolated compounds and standard drug were evaluated in vivo for antimutagenic activity against EMS induced mutagenicity taking mice as model organism by micronucleus and chromosomal aberration tests. Four major compounds were identified as 1) 14, 15-dihydroajugapitin 2) β- Sitosterol 3) Stigmasterol and 4) 8-O-acetylharpagide. A quick and sensitive HPLC method was developed for qualitative and quantitative determination of three isolated marker compounds from Ajuga bracteosa. 14, 15-dihydroajugapitin reduced the micronuclei by 85.10%, followed by β- Sitosterol (72.3%) while as 8-O-acetylharpagide reduced the micronuclei by 46%. It is therefore evident from the present study that the plant contains rich source of anticancer and antimutagenic drugs.

Polarity based characterization of biologically active extracts of Ajuga bracteosa Wall. ex Benth. and RP-HPLC analysis

BackgroundThe concept of botanical therapeutics has revitalized due to wide importance of plant derived pharmaceuticals. Therefore, the ameliorative characteristics of Ajuga bracteosa were studied.MethodsTotal phenolic content, flavonoid content, antioxidant capacity, reducing power and free-radical scavenging activity were determined colorimetrically. Specific polyphenols were quantified by RP-HPLC analysis. Preliminary cytotoxicity was tested using brine shrimp lethality assay while antiproliferative activity against THP-1 and Hep-G2 cell lines was determined by MTT and SRB protocols respectively. Antileishmanial potential was assessed via MTT colorimetric method. To investigate antidiabetic prospect, α-amylase inhibition assay was adopted whereas disc diffusion method was used to detect likely protein kinase inhibitory, antibacterial and antifungal activities.ResultsAmong fifteen different extracts, maximum total phenolic content (10.75 ± 0.70 μg GAE/mg DW), total reducing power (23.90 ± 0.70 μg AAE/mg DW) and total antioxidant capacity (11.30 ± 0.80 μg AAE/mg DW) were exhibited by methanol extract with superlative percent extract recovery (17.50 ± 0.80% w/w). Chloroform-methanol extract demonstrated maximum flavonoid content (4.10 ± 0.40 μg QE/mg DW) and ethanol extract exhibited greatest radical scavenging activity (IC50 14.40 ± 0.20 μg/ml). RP-HPLC based quantification confirmed polyphenols such as pyrocatechol, gallic acid, resorcinol, catechin, chlorogenic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, vanillic acid, coumarin, sinapinic acid, trans-cinnamic acid, rutin, quercetin and kaempferol. The brine shrimp lethality assay ranked 78.60% extracts as cytotoxic (LC50 ≤ 250 μg/ml) whereas significant THP-1 inhibition was shown by methanol-acetone extract (IC50 4.70 ± 0.43 μg/ml). The antiproliferative activity against Hep-G2 hepatoma cancer cell line was demonstrated by n-hexane, ethylacetate and methanol-distilled water (IC50 8.65–8.95 μg/ml) extracts. Methanol extract displayed prominent protein kinase inhibitory activity (MIC 12.5 μg/disc) while n-hexane extract revealed remarkable antileishmanial activity (IC50 4.69 ± 0.01 μg/ml). The antidiabetic potential was confirmed by n-hexane extract (44.70 ± 0.30% α-amylase inhibition at 200 μg/ml concentration) while a moderate antibacterial and antifungal activities were unveiled.ConclusionThe variation in biological spectrum resulted due to use of multiple solvent systems for extraction. We also deduce that the valuable information gathered can be utilized for discovery of anticancer, antileishmanial, antioxidant and antidiabetic bioactive lead candidates.

Antibacterial and antibiofilm potential of leaves extracts of Mirabilis jalapa L. and Ajuga bracteosa wall. against Pseudomonas aeruginosa

In the Present study the antibacterial and antibiofilm potential of Mirabilis jalapa and Ajuga bracteosa crude methanolic extracts of leaves against P1, P2 and P3 clinical strains of Pseudomonas aeruginosa was explored. Antibacterial activity was investigated by agar well diffusion method and the antibiofilm potential was determined by Anti-Pellicle assay and exopolysaccharide inhibition (EI) by Congo red (CR) assay. In antibacterial assay, M. jalapa extract showed good antibacterial activity against P. aeruginosa with maximum 13.5 mm inhibition zone (ZI) against P2 strain and minimum 12.8 mm ZI against P3 strain of P. aeruginosa as compared to A. bracteosa which showed a maximum 13.4 mm ZI and a minimum of 12.7 mm against both P2 and P3 strains of P. aeruginosa, respectively. Both M. Jalapa and A. bracteosa extracts showed moderate (+++) and weak (++++) effect on pellicle inhibition against the tested strains of P. aeruginosa. In CR assay, a moderate antibiofilm effect was recorded on EI for both plants. Based on the results it was concluded that methanol extracts of M. jalapa and A. bracteosa possessed good antibacterial activity and moderate antibiofilm potential against P. aeruginosa. Keywords: Agar well diffusion assay; Ajuga bracteosa; Mirabilis jalapa; Natural products; Opportunistic pathogen http://dx.doi.org/10.19045/bspab.2017.60062

Evaluation of anticonvulsant potential of aqueous methanolic extract and various fractions of Ajuga bracteosa wall

We aim to assess the anticonvulsant potential of Ajuga bracteosa Wall. The aqueous methanol extract (30:70) of Ajuga bracteosa and its n-hexane, chloroform and ethyl acetate fractions were prepared. Anticonvulsant effect was evaluated using different animal models of epilepsy. Crude extract along with its fractions at 500 and 1000 mg/kg doses, administered orally to albino mice were appraised against pentylenetetrazole-induced, strychnine-induced and picrotoxin-induced convulsions. Moreover, effect of test substances on brain GABA level and phenobarbitone induced hypnosis was also studied at dose level of 350 and 400 mg/kg. The plant extract and its fractions produced potent dose dependent anticonvulsant activity in all models of epilepsy. In addition, plant extract and fractions increased brain GABA level and potentiated phenobarbitone-induced sleep duration. From the results it can be deduced that Ajuga bracteosa possess potent anticonvulsant activity, supporting its folkloric use in the treatment of epilepsy.

Metabolic signatures altered by in vitro temperature stress in Ajuga bracteosa Wall. ex. Benth.

To elucidate how biosynthesis of plant metabolites is affected by temperature, metabolite profiles from in vitro regenerated plants raised under different temperature regimes of 10, 15 °C, 20 °C, 25 °C and 30 °C were obtained using electrospray ionization mass spectrometry (ESI-MS), and principal component analysis (PCA) was carried out to identify key metabolites. Several bin masses were detected by PCA loading scatter plots which separated the samples. In-house bin program selectively manifested the putative known metabolites depending on % total ions count and intensity of selected bins in the plant samples. Total phenolic and flavonoid content were harvested to highest levels (12.9 mg GAE/g DW and 9.3 mg QE/g DW), respectively, at 15 °C. Besides, pinoresinol (lignan), some of the vital amino acids such as serine, methionine, histidine and glutamine were found to be at higher amount in plants raised at 15 °C. Significant phenylpropanoids like cinnamic acid, caffeic acid and quercitol were detected at a higher concentration in plants raised at 15 °C as compared to other treatments. However, phosphoenolpyruvate, and oxalosuccinate (intermediates of the pentose phosphate pathway) were accumulated the most in plants raised at 30 °C and they were detected with lowest values at 10 °C. Glucose and deoxy-xylose 5 phosphate (intermediates of TCA cycle) were found in higher amounts at temperature treatments of 15 and 25 °C, respectively. We conclude that a low-temperature treatment (15 °C) results in a stress-induced accumulation of a variety of pharmacologically important secondary metabolites.

Impacts of methyl jasmonate and phenyl acetic acid on biomass accumulation and antioxidant potential in adventitious roots of Ajuga bracteosa Wall ex Benth., a high valued endangered medicinal plant.

Ajuga bracteosa is a medicinally important plant globally used in the folk medicine against many serious ailments. In the present study, effects of two significant elicitors, methyl jasmonate (Me-J) and phenyl acetic acid (PAA) were studied on growth parameters, secondary metabolites production, and antioxidant potential in adventitious root suspension cultures of A. bracteosa. The results showed a substantial increase in biomass accumulation, exhibiting longer log phases of cultures growth in response to elicitor treatments, in comparison to control. Maximum dry biomass formation (8.88DWg/L) was recorded on 32nd day in log phase of culture when 0.6mg/L Me-Jwas applied; however, PAA at 1.2mg/L produced maximum biomass (8.24DWg/L) on day 40 of culture. Furthermore, we observed the elicitors-induced enhancement in phenolic content (total phenolic content), flavonoid content (total flavonoid content) and antioxidant activity (free radical scavenging activity) in root suspension cultures of A. bracteosa. Application of 0.6mg/L and 1.2mg/L of Me-J, root cultures accumulated higher TPC levels(3.6 mg GAE/g DW) and (3.7 mg GAE/g DW) in the log phase and stationary phase, respectively, while 2.5mg/L Me-J produced lower levels (1.4 mg GAE/g DW)in stationary phase of growth stages. Moreover, TFC and FRSA values were found in correspondence to TPC values in the respective growth phases at the similar elicitor treatment. Thus, a feasible protocol for establishment of adventitious roots in A. bracteosa was developed and enhancement in biomass and metabolite content in adventitious root was promoted through elicitation.

Antibacterial activity of 14, 15-dihydroajugapitin and 8-o-acetylharpagide isolated from Ajuga bracteosa Wall ex. Benth against human pathogenic bacteria.

Ajuga bracteosa Wall ex. Benth. (Lamiaceae) commonly known as Bungle Weed has been in use since ancient times and is mentioned Ayurvedic literature. The upper ground parts of the plant are used for treatment of various diseases. The weed is credited with astringent, febrifugal, stimulant, aperient, tonic, diuretic and depurative properties and is used for the treatment of gout and rheumatism, palsy and amenorrhoea. Two compounds 1) 14, 15-dihydroajugapitin and 2) 8-o-acetylharpagide were isolated from the aerial parts of the plant and tested for antibacterial activity against various pathogenic bacteria by agar well diffusion method. Compound 1 and 2 showed maximum antibacterial activity against Escherichia coli with zone of inhibitions of 25. 0±1.4mm and 22.6±0.9mm respectively. The MIC value of compound 1 and 2 ranged between 500 and 1000μg/ml. It could be concluded that both compounds isolated from the aerial parts of Ajuga bracteosa possess antibacterial activity against pathogens.

Comprehensive screening of influential factors in the Agrobacterium tumefaciens-mediated transformation of the Himalayan elixir: Ajuga bracteosa Wall. ex. Benth

Ajuga bracteosa is a valuable medicinal plant producing many important compounds, including withanolides, phytoecdysteroids, neo-clerodane-di and triterpenes, iridoid glycosides, flavonoids, etc. These compounds possess a broad spectrum of biological, pharmacological and medicinal properties, but their yield is very low in wild-type plants, and chemical synthesis is not viable. Metabolic engineering strategies offer a promising solution for the bulk production of these natural products. However, these strategies usually require the establishment of an efficient genetic transformation method, which to date has not been reported for A. bracteosa. Therefore, the current study was conducted by employing Agrobacterium tumefaciens C58C1 harboring the binary plasmid p35SGUSint with GUS as the reporter gene and the NTPII gene as the selectable marker. The influential factors, e.g. explant type, optical density of agrobacterial culture, inoculation period, co-cultivation duration and acetosyringone concentration, were systematically investigated and the optimal transformation conditions were established by 72 independent transformation experiments. Putative transformants were selected on kanamycin 100mg/L. MS medium supplemented with 3.6mg/L BA resulted in maximum regeneration frequency of the transformed explants. The expression of the GUS gene was confirmed by histochemistry and polymerase chain reaction. We established that nodal explants of A. bracteosa precultured for 3days, inoculated with a culture of A. tumefaciens with an OD of 1.0 for 20min and co-cultivated for 2days in MS medium with 200μM acetosyringone at media pH 5.8 showed 100% transformation induction. This is the first report of a successful A. tumefaciens-mediated transformation of A. bracteosa.

GC-MS Analysis and Evaluation of Mutagenic and Antimutagenic Activity of Ethyl Acetate Extract of Ajuga bracteosa Wall ex. Benth: An Endemic Medicinal Plant of Kashmir Himalaya, India

Herbal medicines as the major remedy in traditional system of medicine have been used in medical practices since antiquity. The natural products remain an important source of new drugs, new drug leads and new chemical entities. The ethyl acetate extract of Ajuga bracteosa was evaluated for mutagenic and antimutagenic assay against mice pre-treated with 1/4th LD50 (117.5 mg/kg bw) of ethyl methane sulphonate by micronucleus and chromosomal aberration assay. Mice were treated with ethyl acetate extract of Ajuga bracteosa (Ab-EAE) (100, 200 300 & 400 mg/kg bw) for 30 days. Without the doses of EMS, no mutagenic effects were observed in blood and bone marrow samples of the mice. But Ab-EAE showed antimutagenic effects on EMS induced mutagenicity in mice. It was observed that high doses of Ab-EAE showed protective effects. The reduction profiles in the EMS induction MN at concentration of ethyl acetate extract of Ajuga bracteosa (100, 200, 300 and 400 mg/kg bw) were estimated as 2%, 4.9%, 16.4% and 20.7% respectively. It can be concluded from the study that ethyl acetate extract of Ajuga bracteosa exhibited no mutagenic effects but only possessing antimutagenic effects. This antimutagenic activity is an induction of medicinal relevance.

Callus induction and multiple shoot regeneration in Ajuga bracteosa Wall ex. Benth.-An important medicinal plant growing in Kashmir Himalaya

Ajuga bracteosa Wall ex. Benth is an important medicinal plant growing in Kashmir Himalaya. During the present study an efficient and rapid in vitro protocol has been established viz; Callus induction and multiple shoot regeneration. Callus induction has been achieved from different explants viz., leaf, petiole and internodal cuttings. Maximum callus production was obtained when leaf explants were inoculated on Murashige and Skoog (1962) medium supplemented with BAP (benzylaminopurine 5.0 mg/l) after 19 days of inoculation. In the petiole explant callus was induced on MS (Murashige and Skoog ) medium supplemented with BAP (2.0 mg/l) + IAA ( Indole-3-acetic acid 3.0 mg/l) after 51 days of inoculation and from internodal explant callus was induced on MS medium supplemented with BAP (2.0 mg/l) + NAA(α-Naphthalene acetic acid 5.0 mg/l) after 35 days of inoculation. Callus derived from leaf explants differentiated into multiple shoots on MS medium supplemented with different concentrations of auxins (IAA, NAA) and cytokinins (BAP, Kinetin). Maximum multiple shoots regenerated on MS medium supplemented with BAP (5.0 mg/l) after 28 days of inoculation.

Seasonal and geographical impact on the morphology and 20-hydroxyecdysone content in different tissue types of wild Ajuga bracteosa Wall. ex Benth.

Ajuga bracteosa is an endangered medicinal herb which contains several natural products of therapeutic importance like 20-hydroxyecdysone (20-HE). As geography and habitat play a crucial role in the metabolism and morphology of a plant, the present study was aimed at evaluating the impact of phytogeography, season and tissue type on morphology and 20-HE content of A. bracteosa. The results revealed large morphological variations in various ecotypes of A. bracteosa. However, plants from the same altitude, regardless of their phytogeography, represented similar morphology. Effect of habitat on 20-HE content remained non-significant except for Karot (1608μg/g) and Kahuta (728μg/g). Effect of tissue types was significant (p value <0.016) for 20-HE content and followed ascending order: root<stem<leaf<flower, representing the tender aerial tissues’ hormonal supremacy. Seasons showed a significant impact (p value <0.001) on 20-HE content with the pattern: winter (1902μg/g)>spring (1071μg/g)>summer (617μg/g). The aerial tissue types contained more 20-HE content in all seasons; especially during winter its amount radically rose in flowers (μ=2814μg/g). The aerial portion of Karot ecotype harvested in winter offers a valuable source of 20-HE. To confirm the effect of low temperature on 20-HE content, profiling of A. bracteosa raised in vitro at different temperature regime was carried out. On the basis of these results we hypothesize that chilling cold hampers vegetative growth and triggers stress induced 20-HE accumulation as a defense response.

In-vitro cytotoxicity study of methanolic fraction from Ajuga Bracteosa wall ex. benth on MCF-7 breast adenocarcinoma and hep-2 larynx carcinoma cell lines

Objective:Ajuga bracteosa Wall ex Benth (Labiatae) is popularly known in India as Neelkanthi. A decoction of the leaves, flowers, and barks is used in India for the treatment of cancer including diabetes, malaria, and inflammation etc. The main objective of this study is to investigate the cytotoxic potential of Ajuga bracteosa.Materials and Methods:Successive solvent extraction of Ajuga bracteosa in petroleum ether, methanol, and water extracts was done. These extracts were tested against human breast adenocarcinoma (MCF-7) and larynx carcinoma (Hep-2) tumor cell lines, using the thiazolyl blue test (MTT) assay.Results:The methanolic fraction of Ajuga bracteosa had shown the significant results against MCF-7 and Hep-2 tumor cell lines. The methanolic, petroleum ether and aqueous extract from Ajuga bracteosa, presented an IC50 value at 24 h of 10, 65, 70 μg/ml and 5, 30, 15 μg/ml on MCF-7 and Hep-2 cells, respectively. Steroids compounds namely β-sitosterol and unknown constituents were identified in the most active methanol extract of Ajuga bracteosa wall ex Benth. These known and unknown compounds exhibited cytotoxic potential against MCF-7 and Hep-2 cancer cells.Conclusion:Among all the tested extracts, methanolic extract can be considered as potential sources of anti-cancer compounds. Further studies are necessary for more extensive biological evaluations.