Bioassay guided fractionation and identification of active anti-inflammatory constituent from Delonix elata flowers using RAW 264.7 cells.

CONTEXT: Delonix elata (L.) Gamble (Fabaceae) has been used in the Indian traditional medicine system to treat rheumatism and inflammation. AIM: To assess the anti-inflammatory effect of Delonix elata flowers and to isolate the active principle. MATERIALS AND METHODS: The prompt anti-inflammatory constituent was isolated from Delonix elata flower extracts using bioassay guided fractionation in liposaccharide (LPS) stimulated RAW 264.7 macrophage cell line. The anti-inflammatory activity of extracts/fractions/sub-fractions/compounds (10, 25, and 50 µg/ml) was evaluated by estimating the levels of nitric oxide (NO), TNF-α, and IL-1ß after 24 h of LPS induction (1 µg/ml). The isolated active compound was subjected to NMR, IR, and UV analyses for structure determination. RESULTS: In an attempt to search for anti-inflammatory constituents, the active pure principle was isolated and crystallized as a white compound from Delonix elata flowers methanol extract. This active compound (50 µg/ml) decreased the release of inflammatory mediators levels such as NO (0.263 ± 0.03 µM), TNFα (160.20 ± 17.57 pg/ml), and IL-1ß (285.79 ± 15.16 pg/ml) significantly (p < 0.05); when compared to the levels of NO (0.774 ± 0.08 µM), TNFα (501.71 ± 25.14 pg/ml), and IL-1ß (712.68 ± 52.25 pg/ml) from LPS-stimulated macrophage cells. The active compound was confirmed as hesperidin with NMR, IR, and UV spectroscopy data. This is the first report of this compound from Delonix elata flowers. CONCLUSION: The findings of the study support the traditional use of Delonix elata flowers to treat inflammation.

Traditionally practiced medicinal plant extracts inhibit the ergosterol biosynthesis of clinically isolated dermatophytic pathogens.

OBJECTIVE: Dermatophytes are resistant to some available antibiotics. Development of new plant drugs to control drug resistant microbes is urgently needed. This study evaluates the antidermatophytic potential of 18 selected medicinal plants used by traditional healers in Theni and Virudhunagar Districts of Tamil Nadu, India. MATERIALS AND METHODS: Selected plant parts were collected, shade dried and powdered. Plant powders were extracted with ethanol and their antifungal potency was investigated against and clinical dermatophytes. The antioxidant effect of the extracts was screened using DPPH assay. Minimal Inhibitory Concentration (MIC) and Minimal Fungicidal Concentration (MFC) were estimated for the extracts. Ten plant extracts showed maximum MFC and they were selected to study their efficacy in interfering with ergosterol biosynthesis. Fluconazole-35µg/mL known fungicide was used as control. The most active extracts were taken for biocompatibility studies using 3T3-L1 fibroblast cell lines. RESULTS: The ethanol extract of Phyllanthus reticulates leaves showed good antifungal activity compared to other plant extracts. The MIC and MFC for Phyllanthus reticulatus were 62.5 and 250µg/mL against M. pachydermatitis and T. rubrum respectively. The ethanol extracts of Phyllanthus reticulatus, Coldenia procumbens, Thespesia populnea and Senna alata significantly lowered the release of ergosterol by 16.37, 19.53, 24.79, and 21.44%, respectively. The ethanol extract of Phyllanthus reticulatus leaves was more biocompatible to host cells than other active extracts. CONCLUSION: Our study indicated that the ethanol extract of Phyllanthus reticulates leaves showed promising activity against dermatophytes. It could be a potential material for future development of antidermatophytic agents.

Swertiamarin, a natural steroid, prevent bone erosion by modulating RANKL/RANK/OPG signaling.

Bone erosion is a central feature of rheumatoid arthritis (RA) that is characterized by the infiltration of the synovial lining by osteoclasts and lymphocytes. In the present study, swertiamarin a major secoiridoid glycoside was evaluated for anti-osteoclastogenic property to prevent bone erosion in Freund's complete adjuvant (FCA) induced in-vivo model, in-vitro osteoblast and osteoclasts as well as in co-culture system and in-silico molecular docking analysis. The swertiamarin treatment decreased the expression of TRAP, RANKL, and RANK levels and increased the levels of OPG levels significantly in both in vitro and in vivo models. In in vitro, the compound treatment significantly increased the cell proliferation and ALP levels in osteoblast cells; the high proliferation (153.8600±5.23%) and ALP release (165.6033±4.13%) were observed at 50µg/ml concentration of swertiamarin treatment. At the same time the treatment decreased the TRAP positive cells in osteoclast cells; the high reductions of TRAP positive cells (39.32±3.19%) were observed at 50µg/ml of swertiamarin treatment. The treatment modulated the levels of pro-inflammatory cytokines, MMPs and NF-κB levels in osteoblast and osteoclast co-culture system. In in silico analysis swertiamarin had affinity towards the proteins RANK, RANKL and OPG residues with low binding energy -4.5, -3.92 and -5.77kcal/mol respectively. Thus, the results of this study revealed the anti-osteoclastogenic activity of swertiamarin on the prevention of bone destruction.

Consensus analysis of sastric formulations used by non-institutionally trained siddha medical practitioners of Virudhunagar and Tirunelveli districts of Tamil Nadu, India.

ETHNOPHARMACOLOGICAL RELEVANCE: Siddha system of traditional medicine has been practiced in Tamil Nadu. This system of medicine has a high number of non-institutionally trained practitioners but studies on their traditional medicinal knowledge are not adequate. The present study is aimed to document and analyze the sastric (traditional) formulations used by the non-institutionally trained siddha medical practitioners in Virudhunagar and Tirunelveli districts of Tamil Nadu, India. METHODS: After obtaining prior informed consent, interviews were conducted with 115 non-institutionally trained siddha medical practitioners about the sastric formulations used by them for the treatment. Successive free listing method was adopted to collect the data and the data were analyzed by calculating Informant Consensus Factor (Fic) and Informant Agreement Ratio (IAR). RESULTS: The study documented data regarding 194 sastric formulations and they were classified into plant, mineral and animal based formulations. Quantitative analysis showed that 62.5% of the formulations were plant based, while the mineral based formulations had a high mean number of citations and versatile uses. Gastrointestinal (12.0%), kapha (11.3%) and dermatological (10.8%) ailments had a high percentage of citations. Jaundice had a high Fic value (0.750) followed by the dermatological ailments. The illness categories with high Fic values under each type of formulation were as follows: jaundice, aphrodisiac and urinary ailments (plant based); jaundice, cuts & wounds and dermatological ailments (mineral based); and hemorrhoids, kapha ailments and heart ailments (animal based formulations). The scientific studies conducted with important formulations under each illness category are discussed. CONCLUSION: The present study indicated the importance of some illnesses over the others and inclusion of new illnesses under each formulation. The ingredients used to prepare these formulations have shown varying degrees of scientific evidence; generally limited studies were available on the efficacy of them as formulations. Further in-depth studies on the formulations with high IAR value and Fic value of illness categories will be helpful to improve health status of the people.

Swertiamarin attenuates inflammation mediators via modulating NF-κB/I κB and JAK2/STAT3 transcription factors in adjuvant induced arthritis.

Rheumatoid arthritis (RA) is an autoimmune and chronic inflammatory disease that leads to pannus formation followed by severe joint destruction, characterized by synovial hyperplasia, inflammation and angiogenesis. Swertiamarin is a secoiridoid glycoside that is used as an anti-inflammatory compound, mainly found in Enicostema axillare (Lam) A. Raynal, a medicinal plant used in Indian system of traditional medicine. In the present study, the effect of swertiamarin was evlauated in experimental adjuvant arthritis animal model by the estimation of biochemical (paw thickness, lysosomal enzymes, and urinary degradative products) parameters, proinflammatory cytokines and enzymes along with histopathological and radiographic observations. The proteins of phosphorylated NF-κB/IκB and JAK2/STAT3 transcription factors were also quantified from experimental animals as well as LPS induced RAW 264.7 macrophage cells. In in silico analysis, swertiamarin was docked with proinflammatory enzymes to confirm its potential. The administration of swertiamarin (2, 5, 10mg/kg bw) significantly (P⩽0.05) inhibited the levels of paw thickness, lysosomal enzymes and increased the body weight of experimental animals in a dose dependent manner. In molecular analysis, the treatment decreased the release of proinflammatory cytokines (IL1, TNF, IL-6) and proangiogenic enzymes (MMPs, iNOS, PGE2, PPARγ and COX-2); and also significantly (P⩽0.05) increased the levels of antiinflammatory proteins (IL-10, IL-4) when compared to the disease groups. The swertiamarin treatment significantly (P⩽0.05) inhibited the release of NF-κB p65, p-IκBα, p-JAK2 and p-STAT3 signaling proteins levels on both experimental animals and LPS induced cells. Histopathological and radiological analysis evidenced the curative effect of swertiamarin on bone destruction. The docking studies of swertiamarin on proinflammatory enzymes supported the results from the in vivo experiments. Thus the swertiamarin inhibited the development of arthritis by modulating NF-κB/IκB and JAK2/STAT3 signaling. These findings suggested that swertiamarin acted as an anti-rheumatic agent.

Documentation and quantitative analysis of local ethnozoological knowledge among traditional healers of Theni district, Tamil Nadu, India.

ETHNOPHARMACOLOGICAL RELEVANCE: This study investigated the use of animals among the traditional healers in Theni district of Tamil Nadu, India. The data regarding the medicinal animals/animal products were documented and their usages were analyzed quantitatively. MATERIALS AND METHODS: Based on free list interviews with the traditional healers, we documented the medicinal usage of animals/animal products and calculated the indices such as informant consensus factor (Fic) to determine the consensus over the species for an illness category, as well as the Index Agreement on Remedies (IAR) to determine the extent of potential utilization of each species. RESULTS: In this study, 69 medicinal animals/animal products were documented with the help of standardized questionnaires among the local healers. The results were tabulated and Fic value for each illness category was calculated. Three illness categories viz., jaundice (milk of Capra aegagrus hircus), orthopedics (egg white and meat of Gallus gallus domesticus) and pediatrics (milk of Equus africanus asinus) had got high Fic values. Fifteen illness categories had moderate Fic values. Highly cited animals in these illness categories were: Rusa unicolor (antiemetic), Reticulitermes spp. (diabetes), flesh of Varanus benghalensis (oral ailments), milk (eye ailments, fever) and urine (antidote) of Homo sepians, meat of Trachypithecus johnii (respiratory ailments), various parts of C. aegagrus hircus (blood ailments, coolants, diarrhea, pulmonary and urinary ailments), flesh of Chamaeleon zeyalnica (neural ailments), meat of Passer domesticus (aphrodisiac), curd and dung of Bos primigenius taurus (dermatological ailments), meat of G. domesticus (musculo-skeletal disorders, analgesic), meat of Lissemys punctata (hemorrhoids), and Pherthima posthuma (psychological ailments). Six illness categories had low Fic values. CONCLUSION: This study indicated that the animals are still being used by the local healers of Theni district, to treat various illnesses. Cross-disciplinary approaches to explore the full potential of animal-derived medicines will help to improve the health of local people.

Ameliorative effect of Drynaria quercifolia (L.) J. Sm., an ethnomedicinal plant, in arthritic animals.

Drynaria quercifolia (L.) J. Sm., is an ethnomedicinal plant used widely in Tamil Nadu to treat arthritis. The present study was aimed to evaluate the traditional claim of D. quercifolia rhizome water extract in adjuvant induced arthritic animals. Anti-arthritic effect was studied by assessing the levels of lysosomal enzymes, protein bound carbohydrates, urinary degradative collagen and serum cytokines on control and adjuvant induced arthritis. The paw swelling and body weight were also analyzed. The levels of ROS and lysosomal enzymes in neutrophils of control and adjuvant induced animals were also estimated. D. quercifolia rhizome water extract at doses of 100 and 200mg/kg reduced the paw thickness and elevated the mean body weight of arthritic rats. The treatment with extract showed a significant reduction in the levels of plasma and liver lysosomal enzymes as well as protein bound carbohydrates and urinary degradative collagen levels. The treatment reduced the levels of ROS and lysosomal enzymes in neutrophils significantly. The significant reduction in the levels of serum pro-inflammatory cytokines (TNF-α and IL-1ß) and the increment in the levels of anti-inflammatory cytokine (IL-10) were also observed by the treatment. The present study supports the traditional claim of using D. quercifolia to treat rheumatism.

Quantitative ethnobotany of traditional Siddha medical practitioners from Radhapuram taluk of Tirunelveli District, Tamil Nadu, India.

ETHNOBOTANICAL RELEVANCE: The aim of the present study was to document the knowledge of traditional Siddha medical practitioners from Radhapuram taluk of Tirunelveli district in Tamil Nadu, India, and to quantitatively analyze the data to identify some useful leads for further studies. MATERIALS AND METHODS: Successive free listing was the method adopted for the interview. In this study, 84 traditional Siddha medical practitioners were included and their knowledge on medicinal plants was gathered. The data were assessed with the help of two indices viz., Informant Consensus Factor (F(ic)) and Informant Agreement on Remedies (IAR). RESULTS: The present survey is in accordance with some of the aspects of our previous surveys. Regarding the demography of the informants, it exhibited unevenness in male-female ratio and majority of the informants were poorly educated. Practicing this system of medicine as part time job by majority of the informants might indicate the reduced social status of this medicinal system. The present study had recorded the usage of 217 species which were used to prepare 448 formulations, which in turn yielded 812 use reports. Conversion of use reports had yielded a total of 625 claims and 84.16% of the claims were 'singletons'. The illness category 'adjuvants' holds a high F(ic) value. Among the other illness categories, kapha ailments and dermatological ailments have a high percentage of use-reports. Ailments of blood, jaundice and fever were the other illness categories with high F(ic) values. Some of the claims viz., Mukia madarasepatana (kapha ailments), Mollugo nudicaulis (febrifuge), Indigofera asphalathoides (dermatological ailments), Aerva lanata (urinary ailments), Abutilon indicum (hemorrhoids) and Hybanthus enneaspermus (aphrodisiac), which have relatively high consensus can be taken up for further biomedical studies, since no substantial studies have been conducted on them. CONCLUSION: One of the major aims of National Rural Health Mission is to implement traditional Indian system of medicines into the main stream. In such scenario, scientific validation of community-based local health traditions becomes necessary for their rational implementation. Through this study we have highlighted some claims which are at high use in the study area but having little scientific support. Studies on such claims will provide scientific base which in turn will be useful to improve the health of indigenous people.

Documentation and quantitative analysis of the local knowledge on medicinal plants among traditional Siddha healers in Virudhunagar district of Tamil Nadu, India.

ETHNOPHARMACOLOGICAL RELEVANCE: India has a population with high degree of medical pluralism. Siddha system of Indian traditional medicine is practiced dominantly by the people in Tamil Nadu. The traditionally trained Siddha healers still play an important role in the rural health care. Their knowledge is comparatively more vulnerable than the documented traditional knowledge. Thus, the present study was aimed to document and quantitatively analyze the local knowledge of the traditional Siddha healers in Virudhunagar district of Tamil Nadu, India. MATERIALS AND METHODS: The results presented in this paper are the outcome of series of interviews conducted between January and August, 2010 consisting of 196 field days. After getting prior informed consent, interviews were conducted and successive free-listing was used in the interviews in order to make informants cite the medicinal plants that they have used. By this way 96 healers were interviewed and their data were quantitatively analyzed using various indices such as Informant Consensus Factor (F(ic)), Fidelity Level (FL), Informant Agreement on Remedies (IAR) and Cultural Importance Index (CII). RESULTS: This study recorded the ethno-medicinal usage of 227 species which were used to prepare 611 formulations for the treatment of 36 illness categories. The knowledge holders had the experience of minimum 20 years. There was unevenness in male-female ratio. Regarding the medicinal plants, easily available plants were holding significantly high number of citations, IAR and CII values. Nine illness categories had a high F(ic) value, compared to others. Species with high citations in these groups were Moringa oleifera (aphrodisiacs), Acalypha indica (dermatological ailments), Dodonaea viscosa (musculo-skeletal disorders), Solanum trilobatum (pulmonary ailments), Phyllanthus amarus (jaundice), Piper nigrum (adjuvant) Allium cepa (hemorrhoids), Azadirachta indica (antiseptic) and Tribulus terrestris (urinary ailments). CONCLUSION: Quantitative analysis of the data had revealed that the easily available species hold a high consensus and cultural importance. Future biomedical studies using the medicinal plants enumerated in this study, particularly those with high number of citations and high F(ic) values might yield some novel prototypes. Such studies will also be useful to assess the efficacy and safety of these herbal treatments to take decisions on the health care of rural India.

Consensus of local knowledge on medicinal plants among traditional healers in Mayiladumparai block of Theni District, Tamil Nadu, India.

AIM OF THE STUDY: The role of ethnobotany in drug discovery is huge but there are criticisms over such studies due to their qualitative nature. The present study is aimed at quantitatively abstracting the medicinal plant knowledge of the healers trained in traditional ways, in Mayiladumparai block of Theni District, Tamil Nadu, India. MATERIALS AND METHODS: The interviews and field observations were carried out in all the 18 village panchayaths from January to June 2010, consisting of 148 field days. The interviews were conducted with 80 traditional healers, after obtaining prior informed consent. Successive free listing was used to interview the informants. The informant consensus factor (F(ic)) was calculated to estimate the use variability of medicinal plants. Fidelity index and Cultural importance index were also calculated to analyze the data. RESULTS: This study recorded the ethno-medicinal usage of 142 ethno-species belonging to 62 families that were used to prepare 504 formulations. Jaundice had the highest F(ic) value than all the illness categories studied. Phyllanthus spp. was the highly cited medicinal plant to treat jaundice and had high fidelity index value. This was followed by Senna angustifolia and Terminalia chebula as laxatives. The highly cited medicinal plants in each group with high F(ic) value were Pongamia pinnata (antiseptic), Aerva lanata (antidote and snakebite), Blepharis maderaspatensis (cuts and wounds), Abutilon indicum (hemorrhoids), Ruta graveolens (spiritual medicine), Ocimum tenuiflorum (cough), and Solanum trilobatum (pulmonary ailments). Phyllanthus spp., was the most culturally significant species according to this index, followed by Borassus flebellifer. CONCLUSION: The process of drug discovery has become highly expensive and post-approval and post-marketing withdrawal of drugs is continuing. In such scenario, reverse pharmacology is considered an attractive option. The medicinal plants enumerated in this study with high number of citations and high F(ic) values for illness categories might give some useful leads for further biomedical research.