Synergistic-antagonistic interaction of vegetable extracts, Acalypha indica, Centella asiatica, and Sesbania grandiflora: Wound healing, antioxidant, protectivity, and antimicrobial properties

Aims@#Acalypha indica (AI), Centella asiatica (CA), and Sesbania grandiflora (SG) are vegetables commonly used in traditional medicine in Asian countries to treat skin problems. In this study, we investigated their pharmacological activities relevant to wound healing and synergistic actions to provide an insight into a promising vegetable combination as a candidate treatment for wounds. @*Methodology and results@#The stimulatory, antioxidant, and antibacterial activities of aqueous (A) and methanol (M) extracts of all the three vegetables were assessed alone and in combination in normal human dermal fibroblast (NHDF) cells in vitro. CA-A (89.52%) and the combination of AI-A+CA-A (90.76%) produced the highest percentage of wound closure. AI-A exhibited the highest total phenolic content (TPC) (82.94 mg GAE/g) and moderate reducing activity (61.63 mM Fe (II)/mg) when assessed by ferric reducing antioxidant power (FRAP) assay. Free radical scavenging activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), and the combination of AI-A+CA-A exhibited scavenging activity at IC50 = 379.75 µg/mL and IC50 = 578.7 µg/mL, respectively. Pre-treatment of NHDF cells with CA-M at 100 µg/mL offered the highest protection against hydrogen peroxide. All single and combined vegetable extracts showed poor antibacterial properties against Gram negative and Gram positive bacterial species implicated in wound infection. Only AI-A+CA-A executed synergism in fibroblast migration when assessed via the combination index (CI). Furthermore, screening and identification of AI-A, CA-A, and CA-M via UHPLC (LC-MS/MS) system revealed that the major components responsible for all the tested bioactivities were phenolic groups such as simple polyphenols, flavonoids, polysaccharides, and triterpenes (asiaticoside and madecassosides). @*Conclusion, significance and impact of study@#The vegetable extracts of A. indica, C. asiatica, and S. grandiflora exhibited good bioactivities independently. However, only AI-A+CA-A showed synergism in combination to accelerate the migration of fibroblast and increase antioxidant activities. These findings demonstrate the potential formulation of combined vegetable extracts from the two species of A. indica and C. asiatica for optimum wound healing properties.

Synergistic-antagonistic interaction of traditional herbs Acalypha indica, Centella asiatica, and Sesbania grandiflora: In antifungal-demelanising activities and nitric oxide immunomodulatory responses

Aims@#The oriental-based herbs Acalypha indica (AI), Centella asiatica (CA), and Sesbania grandiflora (SG) possess a broad range of undisclosed therapeutic activities which are edible and easily available throughout the year. To convert the herb extracts into a potential drug form, aqueous (A) and methanol (M) extracts of herbs were assessed alone and in combination for their antifungal-demelanising activity and nitric oxide (NO) immunomodulatory responses. A new bioactive synergistic and antagonistic assessments approach was made on these herbs to identify which extract combination qualifies as a natural drug candidate.@*Methodology and results@#Via micro-dilution technique, methanol extract of A. indica (AI-M) showed the strongest antifungal activity against Aspergillus niger, with a minimum inhibitory concentration (MIC) of 50 mg/mL and a minimum fungicidal concentration (MFC) of 100 mg/mL. Sublethal (50 mg/mL) and subinhibitory (25 mg/mL) doses of AI-M produced the optimal black pigmentation reduction to demelanise A. niger. The combinations AI-M+CA-M, AI-M+SG-M, and CA-M+SG-M showed similar antifungal activities (MIC = 100 mg/mL). At 500 µg/mL, CA-A and the combination CAA+SG-A successfully induced RAW264.7 cells to produce NO at 17.85 µM and 40.84 µM, respectively. The combination of herbs extract showed synergistic interaction towards stimulation of NO production. In contrast, they demonstrated antagonism towards antifungal-demelanising properties. Compound identification of AI-M, SG-M, and SG-A were performed using a UHPLC-QTrap-MS/MS system, which detected phenolic compounds from various groups (cinnamic acids, benzoic acids, and flavonoids).@*Conclusion, significance and impact of study@#The combination of herb extracts showed better stimulation of NO production while the single herb extracts demonstrated good antifungal-demelanising activity. These findings help in the selection of herbs combination for potential natural drug discovery. A good combination of herbs demonstrated synergism to execute better bioactivities compared to individual herb extracts.

An Experimental Evaluation Of Haritamanjari (acalypha Indica Linn) For Mutrala Karma W.s.r To Diuretic Activity

Diuretics are widely employed drugs used to treat extra cellular fluid volume expansion caused due to renal, cardiac, liver disorders etc. These diuretics are effective but have side effects. Acalypha Indica Linn is a herb of Euphorbiaceae family, found throughout India as a weed. Many studies have been carried out but sufficient data is not available to establish its diuretic activity. Hence the study is focused on the assessment of Harita Manjari for its diuretic action in animal model. Methods: The trial drugs are administered for 7 days prior to the day of evaluation of diuretic action; the rats were individually placed in the metabolic cages to collect urine. The volume of the urine collected in graduated vials was measured after 6 hours and expressed in terms of ml/100gm of body weight. Results: The invivo study demonstrated that crude suspension of Harita Manjari in a dose of 450mg/kg has significant diuretic activity. The statistical analysis has been carried out and results on continuous measurements are presented on Mean ± SEM were calculated. Urine output, pH and urine electrolytes were compared with control groups by one-way ANOVA followed by Dunnett’s multiple comparison test. The P Value <0.05 were considered as statistically significant. Conclusion: Order of diuresis: Crude suspension >Alcoholic extract >Aqueous extract.

Determination of Some Minerals and Trace Elements in Medicinal Plants - Acalypha indica (L.), Datura metel (L.) and Tylophora indica Used in the Treatment of Ashtma

In the current era, medicinal plants are the major resources of indigenous medicines in the traditional medicine system and are playing an important role. Over 80% of the world population mainly depend only on traditional/folk medicinal plants for plant-based medicines to control many of the diseases and their extracts for health care. In the present study, the medicinal plants commonly used by the local people and traditional practitioners of Kappathgudda area, Gadag District, North Karnataka region for treatment of Asthma viz. Acalypha indica L., Datura metel L. and Tylophora indica belongs to Euphorbiaceous, Solanaceae and Asclepiadaceae families were selected for elemental analysis due to its wider application in the traditional medicinal system. Analytical Atomic Absorption Spectrophotometer technique was adopted for the determination of various elements and indicated that Fe, Ca, K, Mg, Ti, Mn, V, Zn, Cu and Cd were present in all samples. The obtained results also revealed that the mineral and trace elemental contents were well within the permissible range and hence the medicinal plants are safe to be utilized in the drug formulations.

Acalypha indica root extract increases post-hypoxic rat hippocampal tissue culture cell viability via phospholipase A2 inhibition.

Background: Phospholipase A2 (PLA2) is involved in inflammation and cell death following stroke, and inhibition of its activity may promote neuroregeneration. This study aimed to observe the influence of Acalypha indica Linn root extract towards relative cell viability and PLA2 enzyme level in post-hypoxic hippocampal tissue culture. Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from Sprague Dawley rat exposed to hypoxia with 5% O2 / 5% CO2 / N2 balanced gas for 24 hours. Post-hypoxia, Acalypha indica Linn root extract was added at doses of 10, 15, and 20 mg/mL to three treatment groups. No treatment was given to the control group. Each group consists of six samples. After 72 hours of incubation, relative cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and phospholipase A2 enzyme level was determined using ELISA. Results: PLA2 enzyme level of rat hippocampal tissue culture treated with Acalypha indica Linn root extract at 10, 15, and 20 mg/mL were significantly lower than that of control (5.55 ng/mL, 6.85 ng/mL, and 7.42 ng/mL vs 7.96 ng/mL, p < 0.05). Conclusion: Acalypha indica Linn root extract increases the relative cell viability and decreases the PLA2 enzyme level of post-hypoxic mouse hippocampal tissue with the optimal dose of the extract at 10 mg/mL.

Acalypha indica Linn root extract improved hippocampal cell viability and increased brain-derived neurotrophic factor (BDNF) in hypoxic condition.

Background: This study was done to determine the effect of root extract of Acalypha indica Linn (akar kucing) in protecting neuron viability of the rat hippocampus on tissue culture in hypoxic condition. Methods: This is an experimental study of in vitro primary cell culture of hippocampus of Sprague Dowley adult rat. The cultures were group into control (C) and exposure to root extract of Acalypha indica Linn with dose of 10 mg/mL, 15 mg/mL, and 20 mg/mL for 72 hours. The cultures were then exposed to hypoxic gas (5% oxygen, 5% carbondioxide, nitrogen balance) for 24 hours. After that, relative cell viability was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), cell proliferation by 5-bromo2’-deoxy-uridine (BrdU), and Brain-derived Neurotrophic Factor (BDNF) levels by BDNF ELISA kit. Results: The result showed MTT viability (C: 99.7%, A indica L 10: 326.3%, A indica L 15: 411.7%, A indica L 20: 445.9%), BrdU absorbance (C: 0.07, A indica L 10: 0.10, A indica L 15: 0.12, A indica L 20: 0.13) of the exposured hippocampal cell were significantly higher than the control group (p < 0.01) accompany by increased level of BDNF (C: 11.3 pg/mL, A indica L 10: 12.5 pg/mL, A indica L 15: 23.1 pg/mL, A indica L 20: 18.1 pg/mL). Conclusion: The root extract of Acalypha indica Linn is able to improve rat hippocampal cell viability and endogenous BDNF levels in hypoxic condition.

Phytochemical screening of acalypha Indica l. leaf extracts.

Medicinal plants have been used for centuries as remedies for human diseases because they contain components of therapeutic value. Acalypha indica L. belonging to the family Euphorbiaceae is a popular plant in traditional medicine used for treating various ailments. Phytochemical screening of the leaf extracts of A.indica was carried out following the standard procedures. Aqueous and ethanolic extracts of A.indica leaves indicated the occurence of saponins, flavonoids, terpenoids and cardiac glycosides. The ethanolic extract of A.indica leaves also showed the presence of tannins and steroids. The medical significance of various phytochemical constituents identified in the leaf extracts of A.indica and their potential antimicrobial and therapeutic applications are discussed.

In vitro antiplasmodial effect of ethanolic extracts of coastal medicinal plants along Palk Strait against Plasmodium falciparum

Objective: To identify the possible antiplasmodial compounds from Achyranthes aspera (A. aspera), Acalypha indica (A. indica), Jatropha glandulifera (J. glandulifera) and Phyllanthusamarus (P. amarus). Methods: The A. aspera, A. indica, J. glandulifera and P. amarus were collected along Palk Strait and the extraction was carried out in ethanol. The filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125 μg/mL) of leaf, stem, root and flower extracts of A. aspera, A. indica, J. glandulifera and P. amarus were tested for antiplasmodial activity against Plasmodiumfalciparum. The potential extracts were also tested for their phytochemical constituents. Results:Of the selected plants species parts, the stem extract of A. indica showed excellent antiplasmodial activity (IC50= 43.81μg/mL) followed by stem extract of J. glandulifera (IC50= 49.14μg/mL). The stem extract of A. aspera, leaf and root extracts of A. indica, leaf, root and seed extracts of J.glandulifera and leaf and stem extracts of P. amarus showed IC 50 values between 50 and 100 μg/mL. Statistical analysis revealed that, significant antiplasmodial activity (P<0.01) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out and it showed that there were no morphological changes in erythrocytes by the ethanolic extract of all the tested plant extracts. The in vitro antiplasmodial activity might be due to the presence of alkaloids, glycosides, flavonoids, phenols, saponins, triterpenoids, proteins, and tannins in the ethanolic extracts of tested plants. Conclusions: The ethanolic stem extracts of P. amarus and J. glandulifera possess lead compounds for the development of antiplasmodial drugs.

Increased cell viability and proliferation in post-hypoxic hippocampal tissue culture treated with Acalypha indica root extract.

Background: This research was done to study the influence of Acalypha indica Linn root extract towards relative cell viability and proliferation as parameters of neurogenesis in post-hypoxic hippocampal tissue culture. Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from adult Sprague Dawley rat exposed to hypoxia with 5% O2/5% CO2/N2 balance gas for 24 hours. Post-hypoxia, Acalypha indica Linn root extract was added at doses of 10, 15, and 20 mg/mL to 3 treatment groups. No treatment was given to the control group. Each group consists of 6 samples. After 90 hours of incubation, relative cell viability was measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and cell proliferation was measured by using 5-bromo2’-deoxy-uridine (BrdU) for cell proliferation. Data was analyzed using one way ANOVA parametric tests, then further analyzed with post-hoc analysis. Results: The relative cell viability of rat hippocampal tissue culture treated with Acalypha indica Linn root extract with dose of 10, 15, and 20 mg/mL was significantly higher than control (176.95%, 220.62%, and 386.02% vs. 100%). Cell proliferation of rat hippocampal tissue culture treated with Acalypha indica Linn root extract with dose of 10, 15, and 20 mg/mL was significantly higher than control (0.132, 0.117, 0.114 vs 0.096). Conclusion: Acalypha indica Linn root extract with doses of 10, 15, and 20 mg/mL can increase relative cell viability and proliferation in post-hypoxic hippocampal tissue culture.

Anti-tuberculosis activity of selected medicinal plants against multi-drug resistant Mycobacterium tuberculosis isolates.

Background & objectives: Emergence of multi-drug resistant (MDR) and extensively-drug resistant (XDR) strains of Mycobacterium tuberculosis has further complicated the problem of tuberculosis (TB) control. Medicinal plants offer a hope for developing alternate medicines for the treatment of TB. The present study was done to evaluate in vitro anti-tubercular activity of five medicinal plants viz., Acalypha indica, Adhatoda vasica, Allium cepa, Allium sativum and Aloe vera. Methods: Aqueous extracts of leaves of A. indica, A. vasica, bulbs of A. cepa, cloves of A. sativum and pure gel of A. vera leaves, were tested in vitro for their activity against two MDR isolates (DKU-156 and JAL-1236), reference susceptible strain M. tuberculosis H37Rv as well as rapid grower mycobacterial pathogen M. fortuitum (TMC-1529) using Lowenstein Jensen (L-J) medium and colorimetric BacT/ALERT 3D system. Activity in L-J medium was evaluated by percentage inhibition which was calculated by mean reduction in number of colonies on extract containing as compared to extract free controls. Results: Extracts of all the five plants A. indica, A. vasica, A. cepa, A. sativum and A. vera exhibited anti-tuberculosis activity in L-J medium, the proportion of inhibition of these plants extract in respect mentioned above is 95, 32, 37, 72, 32 per cent, respectively for MDR isolate DKU-156 and 68, 86, 79, 72, 85 per cent, respectively for another MDR isolate JAL-1236, while for sensitive M. tuberculosis H37Rv, inhibition was found to be 68, 70, 35, 63 and 41 per cent, at 4 per cent v/v concentration in L-J medium. There was no inhibition against rapid grower M. fortuitum (TMC-1529). In BacT/ALERT also, extracts of these plants showed significant inhibition against M. tuberculosis. Interpretation & conclusions: Our findings showed that all these plants exhibited activity against MDR isolates of M. tuberculosis. While the anti-TB activity of A. vera, A. vasica and A. sativum against MDR isolates confirm earlier results, activity of the extracts of A. indica and A. cepa is reported for the first time. Further studies aimed at isolation and identification of active substances from the extracts which exhibited promising activities, need to be carried out.

The nerve protection and in vivo therapeutic effect of Acalypha indica extract in frogs.

Aim To demonstrate nerve protection and/or treatment effect of Acalypha indica Linn. extract on nerve paralysis induced by subcutaneus injection of pancuronium bromide on frog’s back. Methods The study was performed on sixty frogs (Bufo melanostictus Schneider) that divided into two groups, i.e. the neuro-protection and neuro-therapy group. Each group was divided further into 6 sub-treatment groups: negative control group treated by water and positive control group treated by piracetam, treatment groups received the extracts 200, 300, 400, 500 mg/kgBW. Pancuronium bromide 0.2% (1 : 20 dilutions) were injected subcutaneously as muscle relaxant. The protective effect was studied by giving the extract orally, 1 hour prior to injection; while the therapeutic effect of the extract was studied by 10 minute treatment after injecting pancuronium bromide solution. The parameters measured were the onset and duration of paralysis (in minutes) and the recovery time (time needed to recover into normal condition). Results The study showed signifi cantly different protective effect of Acalypha indica Linn. root water extract at 400 and 500 mg/Kg.BW compared to negative control group and positive control group (piracetam (p<0.05); while the therapeutic effect was obvious at the dose 200-500 mg/Kg.BW compared to negative control group (p=0.000). There was no signifi cant difference compared to positive control group (piracetam), except at 300 mg/Kg.BW (p=0,012). Conclusion These results have proven that the water extract of Acalypha indica Linn. root has comparable protective and treatment effect on nerves system, as piracetam, but further studies should be performed to provide more evidences particularly pharmacokinetic and pharmacodynamic studies on two animal models that commonly used.