In vitro growth inhibitory effect of selected 18ß-glycyrrhetinic acid esters on Theileriaannulata.

Glycyrrhetinic acid (GA) is one of the important Pentacyclic Triterpenoids (PT) found in the roots of licorice. This study aimed to evaluate the in vitro growth inhibitory effect of 18ß-GA (18ß-Glycyrrhetinic acid) and C-30 esters against Theileria annulata, the causative agent of Tropical Bovine Theileriosis. C-30 esters of 18ß-GA were synthesized and their structures were elucidated using spectroscopy. The pharmacodynamic properties of 18ß-GA and its C-30 esters were predicted using DataWarrior and Swiss ADME tools. Cattle isolates of T. annulata schizont-infected bovine lymphoblastoid cells were cultured using standard conditions and the growth inhibitory effect of GA and its esters were evaluated using MTT assay. The isopropyl ester of 18ß-GA (GI50- 1.638 µM; R2- 0.818) showed improved anti-theileriosis efficacy than other 18ß-GA derivatives. The propyl (GI50 - 5.549 µM), ethyl (GI50 - 5.638 µM), and benzyl (GI50 - 7.431 µM) esters also showed considerable inhibitory effect. The GI50 value for 18ß-GA was recorded as 6.829 µM. This study throws light on the usefulness of 18ß-GA and its esters for the treatment of Tropical Bovine Theileriosis.

Quantitative analysis of medicinal plants used to treat musculoskeletal ailments by non-institutionally trained siddha practitioners of Virudhunagar district, Tamil Nadu, India.

BACKGROUND: Siddha is one of the traditional medical systems of India; previous ethnobotanical survey in Virudhunagar district indicated a high consensus for treating musculoskeletal ailments. OBJECTIVES: This study was aimed to quantitatively document the medicinal plants used to treat musculoskeletal ailments by non-institutionally trained siddha practitioners of Virudhunagar district, Tamil Nadu, India. MATERIALS AND METHODS: This work was the outcome of free-list interviews conducted among 45 informants between April 2016 and January 2017. Sampling sufficiency was assessed by plotting Shannon-Wiener's index and cumulative number of UR. Informant consensus was assessed using Informant Consensus Factor (Fic) and Informant Agreement Ratio (IAR). RESULTS: This study recorded the data regarding 116 plant species which were used to prepare 129 formulations; analysis of the data yielded 490 UR. Among these, 65.3% of UR dealt with internal applications and 34.6% dealt with external applications. In the case of internally used formulations, pain and general musculoskeletal ailments had high Fic values. In the case of externally used formulations, headache and pain had high Fic values. CONCLUSION: Important species prescribed by the informants to treat various musculoskeletal ailments were: Azima tetracantha, Ricinus communis, Sesamum indicum, Moringa oleifera, Cardiospermum halicacabum (internal application), Calophyllum inophyllum, Justicia adhatoda, Curcuma longa, Calotropis gigantea, Zingiber officinale, Withania somnifera, Strychnos nux-vomica, Dodonaea viscosa (external application), Azadirachta indica, Clerodendrum phlomidis,Delonix elata, Pergularia daemia and Vitex negundo (internal and external applications). Robust studies on these local claims will help to improve the community healthcare and will yield some novel agents to treat musculoskeletal ailments.

Antihyperlipidemic effect of iridoid glycoside deacetylasperulosidic acid isolated from the seeds of Spermacoce hispida L. - A traditional antiobesity herb.

ETHNOBOTANICAL RELEVANCE: The interest on herbal health supplements for obesity is increasing globally. Our previous ethnobotanical survey in Tiruvallur district, Tamil Nadu, India indicated the use of Spermacoce hispida L. seeds for the treatment of obesity. AIM OF THE STUDY: This study was aimed to validate the traditional claim and to identify the antihyperlipidemic principle in the seeds of Spermacoce hispida using bioassay guided fractionation method. METHODS: Bioassay monitored fractionation of the aqueous extract from Spermacoce hispida seeds was carried out using triton WR 1339 induced hyperlipidemic animals. It yielded deacetylasperulosidic acid (DAA) as the active ingredient. Pharmacokinetic properties of DAA were predicted using DataWarrior and SwissADME tools. In vitro antiobesity and antihyperlipidemic effects of DAA were evaluated in 3T3L1 preadipocytes and HepG2 cells, respectively. The chronic antihyperlipidemic efficacy of DAA was evaluated in high fat diet fed rats. RESULTS: DAA did not show any mutagenic and tumorigenic properties. It bound with PPARα with comparable ligand efficiency as fenofibrate. The treatment with DAA significantly lowered the proliferation of matured adipocytes, but not preadipocytes. The treatment of steatotic HepG2 cells with DAA significantly decreased the LDH leakage by 43.03% (P < 0.05) at 50 µM concentration. In triton WR 1339 induced hyperlipidemic animals, the treatment with 50 mg/kg dose significantly lowered the TC, TG and LDL-c levels by 40.27, 46.00 and 63.65% respectively. In HFD fed animals, the treatment at 10 mg/kg decreased BMI and AC/TC ratio without altering SRBG. It also improved serum lipid, transaminases and phosphatases levels of HFD fed animals. The treatment lowered adipocyte hypertrophy and steatosis of hepatocytes. CONCLUSION: This preliminary report supported the traditional use of Spermacoce hispida for the treatment of obesity. Further detailed investigations on the long term safety, efficacy and molecular mode of action of Spermacoce hispida and DAA will throw more light on their usefulness for the management of obesity.

Hepatoprotective effect of bisbenzylisoquinoline alkaloid tiliamosine from Tiliacora racemosa in high-fat diet/diethylnitrosamine-induced non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis (NASH) is one of the aggressive forms of non-alcoholic fatty liver disease (NAFLD) and is a potential risk factor of HCC. This study reports the curative effect of tiliamosine on NASH. Tiliamosine was isolated from Tiliacora racemosa Colebr. (Menispermaceae) and its structure was confirmed by studying the physical and spectroscopic data. The effects of tiliamsoine on lipid accumulation and lipotoxicity were evaluated using palmitate-oleate induced steatosis in HepG2 cells. The in vivo efficacy of tiliamosine was evaluated using HFD fed, DEN induced non-alcoholic steatohepatitis Wistar rats. In HepG2 cells, tiliamosine did not affect the cell viability up to 100 µM concentration and showed GI25 value of 264.28 µM. The treatment with tiliamsoine significantly lowered the ORO concentration by 44.17% and triglyceride accumulation by 69.32% at 50 µM concentration (P < 0.005). It also reduced the leakage of LDH and transaminases in PO-BSA induced HepG2 cells. The treatment with tiliamsoine significantly decreased the plasma levels of transaminases, phosphatase and LDH (P < 0.05) in HFD-DEN induced steatohepatitis. The histology and the immunohistochemistry of the hepatic sections were in accordance with the biochemical findings. Preliminary molecular analysis indicated that the hepatic FXR expression was upregulated and TNFα expression was downregulated by the treatment with tiliamsoine. This study provided preliminary evidence on the use of tiliamosine for the treatment of NASH.

Curative effect of arjunolic acid from Terminalia arjuna in non-alcoholic fatty liver disease models.

The prevalence of Non Alcoholic Fatty Liver Disease (NAFLD) is increasing globally. Terminalia arjuna W. & Arn. (Combretaceae) is an endemic tree found in India and Sri Lanka and used traditionally for its cardioprotective and hepatoprotective effects. Arjunolic acid (AA) is an oleanane triterpenoid found mainly in the heartwood of T. arjuna. This study was aimed to evaluate the hepatoprotective effect of AA using cellular and rodent models of NAFLD. AA was isolated from the ethyl acetate extract of the heartwood of T. arjuna. The structure of AA was confirmed by physical and spectroscopic data. Steatosis was induced in HepG2 cells using palmitate-oleate mixture and the effects of AA on triglyceride accumulation and lipotoxicity were assessed. In vivo effect of AA on NAFLD was assessed using HFD fed rats. The treatment with AA did not affect the cell viability upto 100 µM and showed GI25 value of 379.9 µM in HepG2 cells. The treatment with AA significantly lowered the ORO concentration by 35.98% and triglyceride accumulation by 66.36% at 50 µM concentration (P < 0.005) compared to the vehicle treated group. The treatment with AA also reduced the leakage of ALT and AST by 61.11 and 48.29% in a significant manner (P < 0.005). The in vivo findings clearly demonstrated that the animals treated with AA at 25 and 50 mg/kg concentrations showed a significant decrease in the levels of transaminases, phosphatase and GGT (P < 0.005). In the liver, the expression of PPARα and FXRα expressions were upregulated, while PPARγ expression was downregulated by the treatment with AA. The liver histology of the animals showed reduction in steatosis and MNC infiltration. These preliminary evidences suggested that AA might be a promising lead to treat NAFLD. Future robust scientific studies on AA will lead to tailoring it for the treatment of NAFLD.

Hepatoprotective effect of lawsone on rifampicin-isoniazid induced hepatotoxicity in in vitro and in vivo models.

The Drug-induced liver injury is one of the common unfavourable impacts, which seriously affects any drug therapy. This study documented the hepatoprotective efficacy of lawsone, the major bioactive naphthoquinone present in Lawsonia inermis L. (Lythraceae) using in vitro and in vivo models. Lawsone was isolated from the leaves of L. inermis and its structure was confirmed using spectroscopic data. In-vitro antioxidant effect of lawsone was evaluated using ABTS assay. Hepatoprotective effect of lawsone was determined with RIF-INH treated HepG2 cells and Wistar rats. Administration of RIF-INH reduced the viability of the HepG2 cells and the treatment with lawsone significantly restored the viability of the cells even at lower concentration (7.5 µM). The other parameters such as the leakage of transaminases and MDA levels were also significantly reduced by the treatment with lawsone. Oral administration of lawsone to the animals did not show any toxicity up to 2 g/kg b.w. concentration. Treatment with lawsone to the RIF-INH administered animals significantly lowered the serum transaminases levels. The ratio of albumin to globulin was improved and the level of bilirubin was lowered. This study indicated the hepatoprotective effect of lawsone; detailed investigations will give deeper understanding of the application of lawsone for hepatoprotection.

A study on food-medicine continuum among the non-institutionally trained siddha practitioners of Tiruvallur district, Tamil Nadu, India.

BACKGROUND: Medicinal properties of the food species are one of the poorly documented and important areas of ethnopharmacology. The present survey quantitatively documented the medicinal foods prescribed by the non-institutionally trained siddha practitioners of Tiruvallur district of Tamil Nadu. METHODS: Field work was carried out between December 2014 and April 2017 using a questionnaire. The illnesses mentioned by the informants were grouped as illness categories on the basis of emic perceptions. Sufficiency of sampling of this survey was assessed by plotting the cumulative number of UR and Shannon-Wiener's index. The indices such as informant consensus factor (FIC), Index of Agreement on Remedies (IAR), and Cultural Food Significance Index (CFSI) were calculated. RESULTS: This study documented 165 medicinal foods used by 82 non-institutionally trained siddha practitioners of Tiruvallur district, and 73.93% of these preparations were plant based. Among the animal taxa, 82.05% were represented by fish taxa. The illness category gastrointestinal ailments is the majorly cited illness category treated with plant-based formulations. The illness categories viz., gastrointestinal ailments, hemorrhoids, and neural ailments had high consensus under the group of plant-based medicinal foods. In animal-based medicinal foods, kapha ailments had gained 23.07% of UR. The illness categories such as bone fractures, male reproductive ailments, blood ailments, and anabolic had high FIC values. CONCLUSIONS: Deeper studies on different dietary cultures of India may help to derive better interpretations on food-medicine continuum. This study identified some important claims such as the use of citron, pomegranate and Solanum americanum (gastrointestinal ailments), Abutilon indicum, onions and elephant foot yam (hemorrhoids), Boerhavia diffusa (urinary ailments), Moringa oleifera (anemia), Aloe vera (gynecological ailments), Eclipta prostrata (liver ailments), ivy gourd (diabetes), citron (hypertension), Centella asiatica (psychological ailments), spade nose shark (lactogogue), reticulate whipray (wheezing and bronchitis), Katelysia opima (impotence), Indian squid (anemia), and Indian oil sardine (anabolic). More studies on these claims will help identify novel functional foods to add to the field of medical nutrition therapy, with traditional brand identity. Robust studies on the documentation of the traditional knowledge on marine resources will yield a good database for various stakeholders and policy makers.

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.

Effect of two andrographolide derivatives on cellular and rodent models of non-alcoholic fatty liver disease.

The prevalence of Non-Alcoholic Fatty Liver Disease (NAFLD) is increasing and there is an increasing interest in natural products to treat NAFLD. This study aimed to evaluate the hepatoprotective effect of andrographolide and two of its derivatives; in one the OH group at C-14 was removed and in the other OH groups at C-3 and C-19 were protected. Andrographolide (AN) was isolated from the aerial parts of Andrographis paniculata Wall. Isoandrographolide (IAN) and 3,19-acetonylidene andrographolide (ANA) were derivatized from AN. Drug likeness of the compounds was studied using DataWarrior. The effect of the compounds in ameliorating hepatic steatosis and lipotoxicity was assessed using palmitate-oleate induced steatotic HepG2 cell lines. In vivo efficacy of the compounds was assessed by using HFD fed rats. IAN showed comparatively high drug score and low irritability than AN. MTT assay indicated that the treatment with IAN had comparatively less toxicity than AN and ANA to HepG2 cells. The treatment with IAN significantly reduced the lipid accumulation and the leakage of LDH and transaminases, while the treatments with AN and ANA did not prohibit the leakage. In the in vivo experiment, the treatment with IAN showed comparatively better hepatoprotection by reducing the serum lipid, transaminases and ALP levels than with AN and ANA. Our results showed that IAN could be a promising lead to treat NAFLD with comparatively low toxicity and improved efficacy.

Antihyperlipidemic and hepatoprotective effects of Gardenin A in cellular and high fat diet fed rodent models.

The gum of Gardenia resinifera Roth., is one of the important drugs used in the Indian system of medicine and a source of unique polymethoxylated flavones. This study was aimed to evaluate the antihyperlipidemic and anti-NAFLD effects of Gardenin A (Gar-A) from G. resinifera gum using in vitro and in vivo models. Gar-A was isolated from G. resinifera gum and was identified on the basis of the physical and spectral data. Toxicity of Gar-A to HepG2 cells was evaluated using MTT assay. The ability of Gar-A to reduce steatosis was assessed using oleate-palmitate induced HepG2 cell lines by estimating the lipid levels by ORO staining and by estimating the intracellular triglyceride content. Effect of Gar-A on amelioration of lipotoxicity was measured by estimating the LDH levels. The doses for in vivo experiments were fixed by Irwin test, between 50 and 100 mg/kg concentrations, through oral route. The acute antihyperlipidemic effect of Gar-A was assessed in Triton WR-1339 induced hyperlipidemic animals. The chronic antihyperlipidemic and anti-NAFLD effects of Gar-A were evaluated in HFD fed rats. In vitro experiments with HepG2 cell line indicated that the cells treated with Gar-A did not show any significant reduction in the viability up to 70 µg/mL concentration. Steatotic HepG2 cells treated with Gar-A showed a significant reduction in lipid accumulation at 2.5-10 µg/mL concentrations. In triton induced hyperlipidemic rats, the treatment significantly reduced the lipid levels at the synthesis phase. The treatment with Gar-A to the HFD fed animals significantly lowered the steatosis and transaminase levels. The other biochemical parameters such as TC, TG, LDL-c, ALP and ACP were also decreased significantly. Treatment with Gar-A significantly lowered the hyperlipidemia and fat accumulation in the liver; detailed molecular investigations are necessary to establish the antihyperlipidemic and hepatoprotective potentials of Gar-A.

Quantitative ethnomedicinal survey of medicinal plants given for cardiometabolic diseases by the non-institutionally trained siddha practitioners of Tiruvallur district, Tamil Nadu, India.

ETHNOBOTANICAL RELEVANCE: The burden of cardiometabolic diseases such as dyslipidemia, hyperglycemia, hypertension, visceral obesity and atherosclerotic cardiovascular diseases and the use of traditional medicine for the management of such diseases are high in India; hence there is a need to document and analyze such therapies. AIM OF THE STUDY: This study documented and analyzed the medicinal plants prescribed for cardiometabolic diseases by the non-institutionally trained siddha practitioners of Tiruvallur district of Tamil Nadu, India. METHODOLOGY: The field survey was conducted between December 2014 to November 2015. Successive free listing assisted with field-walks was used to interview the informants. After assessing the sampling sufficiency using rarefaction curve analysis, indices such as Informant Consensus Factor (Fic) and Index of Agreement on Remedies (IAR) were calculated for the data. The indicators of informant's medicinal plant knowledge such as Shannon's index, equitability index, etc., were regressed with the demographic profile of the informants. RESULTS: For this study 70 non-institutionally trained Siddha medical practitioners were approached; the data from 36 practitioners who were treating cardiometabolic diseases were documented. This study recorded the use of 188 species which were used to prepare 368 formulations to treat illnesses categorized under cardiometabolic diseases. In this, 53.04% claims were singletons. Regression analysis showed that single species dominance was reduced and the diversity of medicinal plants was increased with the increase in the age and experience. Increase in the years of formal education increased the equitability in the uses. The plants such as Nelumbo nucifera Gaertn. (cardiovascular diseases), Allium sativum L. (dyslipidemia), Cuminum cyminum L. (hypertension), Macrotyloma uniflorum Verdc. (obesity) and Azadirachta indica A. Juss. (type 2 diabetes) were the highly cited medicinal plants. CONCLUSION: This survey has identified the plants most commonly used by Siddha practitioners of Tiruvallur district, Tamil Nadu, India for cardiometabolic diseases. The prevalence of chronic, non-communicable metabolic illnesses such as type 2 diabetes, hypertension and obesity are increasing worldwide due to the rapid changes in the lifestyle. These ailments require a life-long care and in such instances, people tend to use complementary therapies in most cases, alongside with conventional therapies. In view of the high use of traditional therapies for treating cardiometabolic illnesses, this study supports the need for more research to evaluate the potential benefits of the treatments and to identify any safety concern.

Effect of active sub-fraction of Ichnocarpus frutescens (L.) R.Br. in the management of obesity.

AIM OF THE STUDY: Ichnocarpus frutescens (L.) R.Br. (Apocynaceae) is used by the traditional healers in India to treat diabetes and hyperlipidemia.This work aims to study the antiobesity effect of the active sub-fraction obtained from the leaves of this plant using in vitro and in vivo models. METHODOLOGY: Active sub-fraction (Sfr3) isolated by column chromatography was analyzed using Gas chromatography and mass spectrometry. Adipogenesis and apoptosis were detected by oil red O and Hoechst-33342 staining, respectively, in 3T3-L1 adipocytes. Expression of specific marker genes involved in lipid metabolism was detected by Reverse Transcriptase Polymerase Chain Reaction in 3T3-L1 adipocytes and adipose tissues of C57 BL/6J mice, fed with high fat diet (HFD) and treated with Sfr3 and fenofibrate for 45 days. RESULTS: The treatment with Sfr3 decreased lipid accumulation and caused apoptosis in adipocytes. In the HFD induced obese animals, serum and liver lipid levels were decreased and a significant decrease in body weight gain was observed dose dependently in 45 days treatment. Histopathological examination showed decrease in adipose mass and liver lipid accumulation. The effect of Sfr3 on the marker genes of adipocytes was significantly regulated at the treated doses both in vitro and in vivo. CONCLUSION: Sfr3 of Ichnocarpus frutescens is effective in the management of obesity through adipocyte apoptosis.

In vivo and in vitro immunomodulatory potential of swertiamarin isolated from Enicostema axillare (Lam.) A. Raynal that acts as an anti-inflammatory agent.

Swertiamarin is a secoiridoid glycoside found in Enicostema axillare (Lam) A. Raynal, a medicinal plant used as a depurative in the Indian system of traditional medicine. The present study evaluated the immunomodulatory activity of isolated swertiamarin. In vivo immunomodulatory activity of swertiamarin (2, 5, and 10 mg/kg b.w.) was evaluated in a model of sheep red blood cells (SRBC) by assessing its effect on organ weight, hemagglutinating antibody titer (HA), plaque-forming cells (PFC), quantitative hemolysis of SRBC, and delayed type hypersensitivity (DTH). In vitro immunomodulatory potential was studied on isolated splenocytes, neutrophils, and peritoneal macrophages. In silico immunomodulatory effects were evaluated by docking of swertiamarin on proinflammatory cytokines to confirm its potential. In in vivo studies, the animals treated with swertiamarin showed a significant (P ≤ 0.05) increase in antibody titer, plaque-forming cells, and also in weight of the thymus and spleen. A decreased response to DTH reaction was recorded with the treatment of swertiamarin. In in vitro studies, treatment with swertiamarin modulated the messenger RNA (mRNA) and protein expression of IFN-γ, IL-10, and IL-4 significantly (P ≤ 0.05) and also favored Th2-mediated response on concanavalin A (Con A)-induced splenocytes. The compound inhibited the release of free radicals significantly (P ≤ 0.05) in phytohemagglutinin (PHA)-induced neutrophils and also ameliorated the mRNA and protein expression of proinflammatory cytokines (TNF-α, IL-1ß, and IL-6) in lipopolysaccharide (LPS)-induced macrophages. In in silico, the best docked pose of swertiamarin with the target proteins (TNF-α, IL-1ß, and IL-6) was confirmed that swertiamarin acted as an anti-inflammatory mediator.

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.

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.

Anti-inflammatory and anti-arthritic effects of 3-hydroxy, 2-methoxy sodium butanoate from the leaves of Clerodendrum phlomidis L.f.

INTRODUCTION: The leaves of Clerodendrum phlomidis L.f. have been used in the Indian traditional system of medicine to treat several inflammatory diseases and arthritis. The aim of the present study was to assess the anti-inflammatory and anti-arthritic activities of the leaves of C. phlomidis and to isolate the active principle by bioactivity guided fractionation. MATERIALS AND METHODS: To find the anti-inflammatory constituents from this plant, fractionations were performed with concurrent bioassays. Carrageenan-induced inflammation and Freund complete adjuvant (FCA)-induced arthritic rat models were used. The anti-inflammatory and anti-arthritic activities of the isolated compound were studied by assessing the histology of the joints, levels of lysosomal enzymes, protein-bound carbohydrates, acute phase protein, etc., in plasma, as well as by estimating the levels and expression of pro-inflammatory cytokines in the joints. RESULTS: Repeated fractionations and bioassays yielded a novel bioactive compound: 3-hydroxy, 2-methoxy-sodium butanoate. Treatment with this compound reduced the paw edema induced by carrageenan and FCA dose dependently. The levels of lysosomal enzymes and protein-bound carbohydrates decreased significantly upon treatment with the compound. The level of plasma acute phase protein was also decreased compared with control animals. Protein levels and mRNA expression of pro-inflammatory cytokines TNF, IL-1 and IL-6 in the joints were decreased significantly in a dose-dependent manner and the histopathological data also added evidence of the anti-arthritic property of the compound. CONCLUSION: The 3-hydroxy,2-methoxy sodium butanoate isolated from plant leaves displays considerable potency in anti-inflammatory action and has a prominent anti-arthritic effect. This is the first report of this natural compound with bioactivity.

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.

Immunomodulatory potential of Enicostema axillare (Lam.) A. Raynal, a traditional medicinal plant.

ETHNOPHARMACOLOGICAL RELEVANCE: Enicostema axillare (Lam.) A. Raynal., (Gentianaceae) has been used in traditional Indian system of medicine as depurative and for the treatment of psoriasis, intermittent fever and cancer. Ethnobotanical survey conducted in Theni District indicated a high consensus for this plant as blood purifier and to treat dermatopathy and venereal infections. The present study evaluated the immunomodulatory activity of the methanol extract of Enicostema axillare. MATERIALS AND METHODS: In vivo immunomodulatory activity of Enicostema axillare methanol extract (100 and 200mg/kg b.w) was evaluated by assessing its effect on the total and differential leukocyte count, organ weight, hemaggultinating antibody titer, plaque forming cells, quantitative hemolysis of SRBC and delayed type hypersensitivity. Sheep red blood cells (SRBC, 5×10(8)cells/0.1mL) were used to immunize the animals. In vitro immunomodulatory potential of the extract was studied using peritoneal macrophages by evaluating its effects on NBT reduction, NO production and cytokine release. RESULTS: The animals treated with Enicostema axillare methanol extract showed a significant (P≤0.05) increase in weight of the thymus and spleen. The total leukocyte and lymphocyte count was increased significantly (P<0.005) by the treatment. There was no significant alteration in neutrophil count. A dose dependent increase in antibody titer value was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen on treatment with the extracts at 10 and 25µg/mL. The extract inhibited the release of pro-inflammatory cytokines and production of NO significantly in a dose dependent manner. CONCLUSION: These findings suggested that the methanol extract of Enicostema axillare acted on both humoral and cell mediated immune functions and decreased the release of pro-inflammatory cytokines in the peritoneal macrophages.

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.