Influence of two natural cross-linkers on microtensile bond strength durability ­ An in vitro study / Influência de dois reticuladores naturais na durabilidade da resistência de união à microtração ­ Um estudo in vitro

Objective: to investigate the effect of two natural cross-linkers on microtensile bond strength (µTBS) and evaluate their influence on the durability of the resin dentin bonds. Material and Methods: the Moringa oleifera and Centella asiatica plant extracts were qualitatively tested with high-performance thin layer chromatography (HPTLC) for the presence of phenols. The phenolic content ranged from 27 to 30 gallic acid equivalents (GAE), µg/mg of dry weight. After etching, two concentrations (5% and 1%) of these two extracts were prepared and used as pretreatment liners on dentin. They were applied for a min. After restoration with resin composite, dentin resin beams were prepared. The study groups were 5% Moringa, 1% Moringa 5% Centella 1% Centella, and control (without cross-linker application). For each group, half of the samples underwent µTBS testing after 24 hours, while the remaining half were immersed in artificial saliva to assess the bond's longevity after 6 months of ageing. Statistical analysis was performed using one-way ANOVA followed by Tukey's post hoc test. Results: both 5% and 1% Moringa showed a significant difference (p<0.05) compared to the other groups at both intervals. However, after ageing, the specimens in the control and 1% Centella groups resulted in a significant decrease in µTBS. Conclusion: overall, both concentrations of Moringa (5% and 1%) were effective in stabilising the bond during both intervals.(AU)

Objetivo: investigar o efeito de dois reticuladores naturais na resistência de união (µTBS) à microtração e avaliar sua influência na durabilidade da adesão da resina à dentina. Material e Métodos: extratos das plantas Moringa oleifera e Centella asiatica foram qualitativamente testados através de cromatografia em camada fina de alta performance (HPTLC) para a presença de fenóis. O conteúdo fenólico alcançou entre 27 a 30 equivalentes de ácido gálico (GAE), µg/mg de peso seco. Após o condicionamento, duas concentrações (5% e 1%) dos extratos foram preparadas e utilizadas como forros de pré-tratamento em dentina. Eles foram aplicados por um minuto. Após a restauração com resina composta, palitos de dentina e resina foram preparados. Os grupos foram 5% Moringa, 1% Moringa, 5% Centella, 1% Centella e controle (sem aplicação de reticulador). Para cada grupo, metade das amostras foram submetidas ao teste µTBS após 24 horas, enquanto a outra metade foi imersa em saliva artificial para avaliar a longevidade adesiva após 6 meses de envelhecimento. Foi realizada análise estatística através de ANOVA 1-fator, seguido do teste post hoc de Tukey. Resultados: ambas as concentrações de 5% e 1% de Moringa demonstraram diferença significativa (p<0.05) comparadas aos outros grupos em ambos os intervalos. No entanto, após o envelhecimento, os espécimes dos geupos controle e 1% de Centella resultaram em uma redução significativa de µTBS. Conclusão: no geral, ambas as concentrações de Moringa (5% e 1%) foram efetivas em estabelecer a adesão em ambos os intervalos (AU)

A standardized extract of Centella asiatica (ECa 233) prevents temporomandibular joint osteoarthritis by modulating the expression of local inflammatory mediators in mice

Abstract Objectives To investigate the effect of a standardized extract of Centella asiatica (ECa 233), which has anti-inflammatory properties, on the local expression of the transient receptor potential vanilloid 1 (TRPV1), the acid-sensing ion channel subunit 3 (ASIC3), and the calcitonin gene-related peptide (CGRP) in the temporomandibular joint (TMJ) structure 21 days after injecting the TMJ with complete Freund's adjuvant (CFA). Methodology A mouse model was induced by analyzing the CFA-injected TMJ on days 7, 14, and 21. We assessed TMJ histology by the osteoarthritis cartilage grade score. Then, we observed the effect of different ECa 233 concentrations (30, 100, and 300 mg/kg) and of 140 mg/kg ibuprofen doses on TRPV1, ASIC3, and CGRP local expression on day 21. Results Osteoarthritis cartilage scores were 1.17±0.37 and 3.83±0.68 on days 14 and 21, respectively, in the CFA group (n=5). On day 21, TRPV1, ASIC3, and CGRP expression significantly increased in the CFA group. In the ibuprofen-treated group, TRPV1 expression significantly decreased, but ASIC3 and CGRP showed no significant difference. All ECa 233 doses reduced TRPV1 expression, but the 100 mg/kg ECa 233 dose significantly decreased ASIC3 expression. Conclusions TRPV1, ASIC3, and CGRP expression increased in mice with TMJ-OA on day 21. All ECa 233 and ibuprofen doses inhibited pathogenesis by modulating the local expression of TRPV1 and ASIC3. Therefore, ECa 233 was more effective than ibuprofen.

The Potential of Centella asiatica (Linn.) Urban as an Anti-Microbial and Immunomodulator Agent: A Review

Centella asiatica (Linn.) Urban (Umbelliferae) which is also known as ‘pegaga’ is highly consumed and eaten raw as ‘ulam’ in Malaysia. C. asiatica is used in traditional medicines to treat various health conditions such as rheumatism, inflammation, syphilis, skin diseases and diarrhoea. Various reports exhibited that the crude extracts and isolated bioactive compounds of C. asiatica possessed a broad range of pharmacological activities such as anti-oxidant, anti-diabetic, anti-tumor, wound healing, anti-microbial, anti-inflammatory, immunomodulatory, hepatoprotective and memory enhancing properties. The pharmacological validation on anti-microbial and immunomodulatory of C. asiatica is very limited and several existence review papers related for this plant had not been focused for both activities. This review therefore attempts to combine the existing literature to offer immense scope for researchers engaged in validation of the traditional claims and bioactivities of this plant related with anti-microbial and immunomodulatory potential.

Combination Effect of Titrated Extract of Centella asiatica and Astaxanthin in a Mouse Model of Phthalic Anhydride-Induced Atopic Dermatitis

PURPOSE: In our previous study, we demonstrated that both titrated extract of Centella asiatica (TECA) and astaxanthin (AST) have anti-inflammatory effects in a 5% phthalic anhydride (PA) mouse model of atopic dermatitis (AD). The increasing prevalence of AD demands new therapeutic approaches for treating the disease. We investigated the therapeutic efficacy of the ointment form of TECA, AST and a TECA + AST combination in a mouse model of AD to see whether a combination of the reduced doses of 2 compounds could have a synergistic effect. METHODS: An AD-like lesion was induced by the topical application of 5% PA to the dorsal ear and back skin of an Hos:HR-1 mouse. After AD induction, TECA (0.5%), AST (0.5%) and the TECA (0.25%) + AST (0.25%) combination ointment (20 μg/cm2) were spread on the dorsum of the ear or back skin 3 times a week for 4 weeks. We evaluated dermatitis severity, histopathological changes and changes in protein expression by Western blotting for inducible nitric oxide synthase (iNOS), cyclocxygenase (COX)-2, and nuclear factor (NF)-κB activity. We also measured the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and immunoglobulin E (IgE) in the blood of AD mice by enzyme-linked immunosorbent assay (ELISA). RESULTS: PA-induced skin morphological changes and ear thickness were significantly reduced by TECA, AST and TECA + AST treatments, but these inhibiting effects were more pronounced in the TECA + AST treatment. TECA, AST and the TECA+AST reatments inhibited the expression of iNOS and COX-2; NF-κB activity; and the release of TNF-α, IL-6 and IgE. However, the TECA+AST treatment showed additive or synergistic effects on AD. CONCLUSIONS: Our results demonstrate that the combination of TECA and AST could be a promising therapeutic agent for AD by inhibiting NF-κB signaling.

Effects of centella asiatica granule on the expression of TGF-β and related down-stream signals in rats with early diabetic nephropathy / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of centella asiatica (CA) granule on the expression of transform growth factor-β(TGF-β) and related down-stream signals in rats with early diabetic nephropathy(DN) and to clarify the molecular mechanisms of CA molecular mechanism of on preventing and curing early diabetic kidney disease DN by studying the effects of centella asiatica on TGF-β expression and related down-stream signals.@*METHODS@#Sixty male SD rats were divided into control group(=10) and DN model group(=50). The model rats were made a right nephrectomy. One week later, diabetic nephropathy was induced by intraperitoneal injection of streptocozin(30 mg/kg) for three consecutive days. High blood glucose level of Tail vein (fasting glucose ≥ 16.7 mmol/L) and high urinary protein level(total protein level in DN group was more than twice higher than the control group) were measured to confirm early DN in rats. In the sham operation group, the right renal capsule was damaged and the corresponding amount of saline was injected. The model rats were administrated by the means of intragastric administration. The DN model group were divided into DN group, DN+fosinopril group(1.6 mg/kg·d), DN+high CA group(16.8 mg/kg·d), DN+medium CA group(11.2 mg/kg·d) and DN+low CA group(5.6 mg/kg·d), and each group was intragastric administration one time every morning last for 16 weeks. The expressions of mRNA and protein of TGF-β, TβR1, TβR2, Smad2/3, Smad7 and the level of Smad2/3 phosphorylation were detected by using real time quantitative polymerase chain reaction and Western blot.@*RESULTS@#The expressions of mRNA and protein of TGF-β, TβR1, TβR2, Smad2/3 and the level of Smad2/3 phosphorylation were significantly increased, the expressions of mRNA and protein of Smad7 were dramatically decreased. The fosinopril and high dosage CA could reverse the effects of DN.@*CONCLUSIONS@#CA plays an important role in preventing and curing DN through regulating the TGF-β/Smad signaling pathways.

Effects of centellaasiatica granule on the expression of Smad 2/3, Smad 7 and collagen Ⅳ in the mesangial cells stably expressed TGF-β1 / 中国应用生理学杂志

OBJECTIVES@#Stably expressed transforming growth factor -beta 1(TGF-β1)MCs were obtained and the effects of centellaasiatica (CA) granule on the expressions of Smad 2/3, Smad 7 and collagen Ⅳ and the level of Smad 2/3 phosphorylation were observed.@*METHODS@#Lipofectin method was used to transfect TGF-β1 vector into MC, and the stably expressed TGF-β1 cell lines were selected by G418. The cells were divided into three groups. Control group:normal MC + RPMI 1640 + 10% normal rat serum; TGF-β1 group:stably expressed TGF-β1 MC + RPMI 1640 + 10% normal rat serum; CA group:stably expressed TGF-β1 MC + RPMI 1640 + 10% rat serum containing high CA. The experiments were repeated for five times. The contents of TGF-β1 and collagen Ⅳ in the culture medium were detected with ELISA, the expressions of mRNA and protein of TGF-β1, Smad 2/3, Smad 7 and the level of Smad 2/3 phosphorylation were detected by using real time quantitative polymerase chain reaction and Western blot.@*RESULTS@#The contents of TGF-β1 and collagen Ⅳ in the culture medium of stably-expressed TGF-β1 MC were increased significantly, and the CA could reverse the effects of TGF-β1. The expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation were increased significantly in TGF-β1 transfected MC, and CA could dramatically reduce the expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation. The high expression of TGF-β1 decreased the expression of Smad 7 mRNA and protein, and the CA could antagonize the effect of mRNA expression.@*CONCLUSIONS@#The MCs stably-expressed TGF-β1 can activate the TGF-β1/Smad signal pathway and increase the expression of collagen Ⅳ. CA can decrease the occurrence of diabetic nephropathy(DN) by reducing the production of collagen Ⅳ through inhibiting the TGF-β1/Smad signal pathway.

Comparative effects of aqueous single-phase and oil-water two-phase mouthrinses containing bamboo salt, magnolia bark and Centella asiatica extracts on reducing gingivitis: a randomized clinical trial / 대한구강보건학회지

OBJECTIVES: The aim of this study was to assess the antiplaque and antigingivitis effectiveness of aqueous single-phase and oil-water two-phase mouthrinses, containing bamboo salt, magnolia bark, and Centella asiatica extracts, in Korean adults. METHODS: In this double-blinded clinical trial, a total of thirty-four participants aged over 19 years were randomly allocated to three experimental groups: 1) control group; 2) aqueous single-phase mouthrinse (ASM) group, and; 3) oil-water two-phase mouthrinse (OTM) group. The experimental mouthrinses all contained sodium fluoride, and the ASM and OTM contained additional ingredients of bamboo salt, magnolia bark, and Centella asiatica extracts. For the OTM, 50% essential oil was added to create an oil-water two-phase mouthrinse. A two-week randomized crossover design with a two-week washout period was applied. Following a complete dental prophylaxis, participants were instructed to use the prescribed mouthrinse twice daily for two weeks as an adjunct to their usual mechanical oral hygiene procedures. Pre- and post-experiment clinical examinations were performed to measure the plaque index (PI) and bleeding on probing (BOP) for the full mouth. Paired t-test was applied to compare the intergroup differences for all clinical variables. RESULTS: Compared to the control group, ASM showed a significantly reduced BOP (P<0.05). However, there was no statistically significant difference in the effects of the three mouthrinses on reducing the PI. CONCLUSIONS: The results of this study indicate that the use of an aqueous, single-phase mouthrinse containing bamboo salt, magnolia bark and Centella asiatica extracts could help alleviate gingivitis.

Recent Updates in Neuroprotective and Neuroregenerative Potential of Centella asiatica

Centella asiatica, locally well known in Malaysia as pegaga, is a traditional herb that has been used widely in Ayurvedic medicine, traditional Chinese medicine, and in the traditional medicine of other Southeast Asian countries including Malaysia. Although consumption of the plant is indicated for various illnesses, its potential neuroprotective properties have been well studied and documented. In addition to past studies, recent studies also discovered and/or reconfirmed that C. asiatica acts as an antioxidant, reducing the effect of oxidative stress in vitro and in vivo. At the in vitro level, C. asiatica promotes dendrite arborisation and elongation, and also protects the neurons from apoptosis. In vivo studies have shown that the whole extract and also individual compounds of C. asiatica have a protective effect against various neurological diseases. Most of the in vivo studies on neuroprotective effects have focused on Alzheimer’s disease, Parkinson’s disease, learning and memory enhancement, neurotoxicity and other mental illnesses such as depression and anxiety, and epilepsy. Recent studies have embarked on finding the molecular mechanism of neuroprotection by C. asiatica extract. However, the capability of C. asiatica in enhancing neuroregeneration has not been studied much and is limited to the regeneration of crushed sciatic nerves and protection from neuronal injury in hypoxia conditions. More studies are still needed to identify the compounds and the mechanism of action of C. asiatica that are particularly involved in neuroprotection and neuroregeneration. Furthermore, the extraction method, biochemical profile and dosage information of the C. asiatica extract need to be standardised to enhance the economic value of this traditional herb and to accelerate the entry of C. asiatica extracts into modern medicine.

Biological Activities and Stability of a Standardized Pentacyclic Triterpene Enriched Centella asiatica Extract

Pentacyclic triterpenes, mainly, asiatic acid, madecassic acid, asiaticoside, and madecassoside are the active constituents of Centella asiatica. A pentacyclic triterpene enriched C. asiatica extract (PRE) was prepared and standardized to contain a total pentacyclic triterpenes not less than 65% w/w. This work was focused on determination of antiinflammatory, antioxidant, and tyrosinase inhibitory activities of PRE and its stability. The PRE exhibited a satisfactory nitric oxide inhibitory effect, with an IC50 value of 64.6 µg/mL. In addition, the PRE inhibited tyrosinase enzyme activity with an IC50 value of 104.8 µg/mL. In contrast, the PRE possessed only weak antioxidant activity. The PRE was stable over a period of four months when stored as a dried powder but only in a well-closed container protected from light at 4 °C. An aqueous alcoholic solution of the PRE was stable at pH values of 5.8 and 7.0, but was not stable at a pH of 8.2. Preparations of the PRE in an aqueous solution should be performed in acidic or neutral conditions.

Lâminas de irrigação e sombreamento na produção de biomassa de Acariçoba (Hydrocotyle umbellata L. ) / Levels of irrigation and shade in the production of the biomass of dollarweed (Hydrocotyle umbellata L. )

RESUMO: A acariçoba é uma planta utilizada pela medicina ayurvédica e em cosméticos por possuir propriedades rejuvenescedoras e revitalizante do sistema nervoso e cérebro. Com objetivo de avaliar os caracteres agronômicos e o teor de flavonóides totais de acariçoba em resposta a aplicação de lâminas distintas de irrigação e sombreamento, foi conduzido um experimento em área localizada nas coordenadas 17º 47' 53'' latitude (S); longitude (W) - 51º 55' 53'', com altitude média de 648m, no período de julho a novembro. O clima é Aw (tropical), relevo suave ondulado (8% de declividade) com solo classificado como Latossolo Vermelho Distroférrico e textura argilosa. As temperaturas médias anuais são 18,1ºC e 30ºC. A umidade relativa do ar média é de 64,7% e a precipitação total anual média é de 1550 mm, com estações do ano seca e chuvosa, bem definidas. O ensaio foi conduzido em blocos casualizados em esquema fatorial 2 x 2, com 5 repetições totalizando vinte unidades experimentais com dois manejos de irrigação, sendo um com aplicação de uma lâmina de água diariamente, e outro, com duas aplicações de lâmina de água diariamente. Os tratamentos de sombreamento foram a pleno sol e com 50% de sombreamento. Foram avaliadas a massa fresca e massa seca das partes aéreas, a altura e o número de plantas, e o teor de flavonóides totais das partes aéreas. A produção de flavonóides foi favorecida pelo sombreamento, enquanto a irrigação não influenciou nem na produção de flavonoides, nem nos caracteres agronômicos avaliados.

ABSTRACT: Dollarweed is a plant that is widely used by the Ayurvedic medicine and in the cosmetics industry because of its rejuvenating and revitalizing properties related to the nervous system and the brain. In order to evaluate the agronomic characteristics and the content of total flavonoids of the dollarweed, in response to the application of different levels of irrigation and shade, an experiment was conducted in an area at the coordinates 17º 47' 53'' latitude (S), 51º 55' 53'' longitude (W), with a mean altitude of 648m, from July to November. The climate is Aw (tropical), gently undulating relief (8% slope) with distroferric red latosol and clayey texture. The average annual temperatures are 18.1 ºC and 30 ºC. The average relative humidity of the air is 64.7% and the average annual rainfall is 1550 mm, with well-defined dry and rainy seasons. The assay was in randomized blocks, with a 2 x 2 factorial design, with 5 repetitions, amounting to twenty experimental units with two irrigation managements, being one with a daily application of water and the other with two daily applications of water. The shading treatments were under full sun and 50% shade. The fresh and dry weights of the aerial parts, the height and number of plants and the total content of flavonoids of the aerial parts were evaluated. The production of flavonoids was favored by the shade; irrigation did not influence the production of flavonoids or the agronomic characters evaluated.

Asiatic acid mitigates hyperglycemia and reduces islet fibrosis in Goto-Kakizaki rat, a spontaneous type 2 diabetic animal model / 中国天然药物

The Goto-Kakizaki (GK) rat is a spontaneous type 2 diabetic animal model, which is characterized by a progressive loss of beta islet cells with fibrosis. In the present study, the hypoglycemic effect of asiatic acid (AA) in GK rats was examined. GK rats receiving AA at a daily dose of 25 mg·kg(-1) for four weeks showed a significant reduction in blood glucose levels. Age-matched normal Wistar rats were given 0.5% sodium carboxymethyl cellulose (CMC-Na) solution for the same periods and used as control. Compared to the normal Wistar rats, GK rats treated with AA showed improvement in insulin resistance partially through decreasing glucose level (P < 0.01) and insulin level (P < 0.05). Furthermore, the results of immunohistochemistry indicate that AA treatment reduced islet fibrosis in GK rats. Fibronectin, a key protein related to islet fibrosis, was over-expressed in GK rats, which was reversed significantly by AA treatment (P < 0.05). These findings suggest that AA has a beneficial effect on lowering blood glucose levels in GK rats and improves fibrosis of islets in diabetes, which may play a role in the prevention of islets dysfunction.

Effect of beta-glucan from Aureobasidium on dermal wound healing in diabetic C57BL/KsJ-db/db mouse model / 동물의과학연구지

The objective of the present study was to examine the effects of beta-glucan originating from Aureobasidium on full-thickness skin wound healing in diabetic C57BL/KsJ-db/ db mouse models. In the diabetic C57BL/KsJ-db/db model, test articles were topically applied twice a day for 20 days starting from 1 day after wounding. The results were compared to that of MadecassolTM ointment (madecassol; 1% Centella asiatica extracts) topically applied at a concentration of 100 mg/kg. Treatment with beta-glucan resulted in significant (p<0.01 or p<0.05) and dose-dependent decreases in wound size compared with that of vehicle control showing increased wound size (WS, %). In addition, 50% contraction time (CT50) was dramatically and dose-dependently reduced, and inflammatory cells in granulation tissues of the wound area were significantly (p<0.01 or p<0.05) and dosedependently reduced compared with that of vehicle control showing increased numbers of micro-vessels and fibroblasts as well as re-epithelialization. In the madecassol group, similar changes in inflammatory cells and fibroblasts with re-epithelialization were also observed, but madecassol did not influence angiogenesis. No meaningful changes in body weight were detected in all tested groups compared with the vehicle control. Therefore, these data suggest that beta-glucan has a beneficial effect on diabetic delayed skin wound healing and may be useful to manage incurable skin wounds in diabetic animals.

Centella asiatica enhances neurogenesis and protects neuronal cells against H2O2-induced oxidative injury / 동물의과학연구지

Traditionally, Centella asiatica leaf extracts are used to treat neurodegenerative diseases in India. Centella asiatica is reportedly used to enhance memory and treat dementia, but its promoting effect on neural stem cell differentiation has not been studied yet. In the present study, we investigated whether or not Centella asiatica leaf extracts act on neuronal precursor cells and neuronal cell lines to induce neuronal differentiation, neurite outgrowth, and neuroprotection. The neurogenesis-promoting potential of Centella asiatica leaf extracts was determined by differentiation assay on neural stem cells isolated from mouse embryos and PC12 cell lines. To understand the contribution of specific neural cell types towards increase after Centella asiatica treatment, neural stem cells were differentiated into various neural subtypes and checked by Western blotting using neural cell lineage-specific antibody markers. Neuroprotective activity of Centella asiatica was analyzed in PC12 cells exposed to 100 microM of H2O2. Cell growth was analyzed by MTT assay while cell death was analyzed by Western blotting detection of apoptosis-related proteins. Cells treated with Centella asiatica had significantly longer primary and secondary neurites as well as a higher number of neurites per cell compared to control cells. Expression levels of TUBBIII, TH, NF, and BDNF increased upon Centella asiatica treatment, suggesting that Centella asiatica has a neurogenesis-promoting effect. Centella asiatica also inhibited oxidative stress-induced neural cell damage through regulation of apoptosis- and cell cycle-related proteins. Thus, leaf extracts of Centella asiatica might promote neurogenesis, neuroregeneration, and neuroprotection in the context of neurodegenerative diseases.

Hypolipidemic and hypoglycemic effects of Centella asiatica (L.) extract in vitro and in vivo.

In vitro study revealed that pancreatic lipase inhibitory activity of C. asiatica extract was significantly higher than rutin but lower than orlistat, an anti-obesity drug. α-Amylase inhibitory activities of C. asiatica extract and rutin were significantly lower than acarbose, an anti-diabetic drug. Inhibition of α-glucosidase activity by C. asiatica extract, rutin, and acarbose was not different. The in vivo study substantiated the in vitro results. C. asiatica extract (1000 and 2000 mg/4 mL/kg), rutin (1000 mg/4 mL/kg), and orlistat (45 mg/4 mL/kg) significantly decreased plasma glucose, triglyceride and total cholesterol levels in lipid emulsion-induced hyperlipidemic rats at 3 h. However, plasma aspartate aminotransferase and alanine aminotransferase levels did not show significant change. The present work further supports that the C. asiatica extract and its bioactive rutin may help managing hypolipidemic and hypoglycemic effects.

A LC-ESI-MS method for the simultaneous determination of madecassoside and its metabolite madecassic acid in rat plasma: comparison pharmacokinetics in normal and collagen-induced arthritic rats / 中国天然药物

To develop a simple and highly sensitive high performance liquid chromatography with electrospray ionization mass spectrometric (LC-ESI-MS) method for the simultaneous determination of madecassoside and its major metabolite madecassic acid in rat plasma, and compare the pharmacokinetics of the two compounds in normal and collagen-induced arthritis (CIA) rats. Glycyrrhetinic acid was used as the internal standard (IS). Chromatographic separation was accomplished on an Inertsil ODS-3 column, using a gradient elution with the mobile phase composed of acetonitrile and water acidified with 0.1% (V/V) formic acid. Detection was achieved by ESI-MS under the negative selected ion monitoring (SIM) mode. In normal and CIA rats, madecassoside (30 mg·kg(-1)) was orally administered for 21 consecutive days from the day of arthritis onset. For madecassoside, the linear range was 10-1 000 ng·mL(-1) with the square regression coefficient (r) of 0.998 9, while for madecassic acid, the linear range was 10-500 ng·mL(-1) with the square regression coefficient (r) of 0.996 1. The lower limit of quantification was 10 ng·mL(-1) for both analytes. The intra- and inter-day precision ranged from 1.78% to 13.42% for madecassoside and 2.30% to 14.90% for madecassic acid, and the accuracy was between -0.95% and 6.30% for madecassoside and between -1.48% and 5.34% for madecassic acid. The average recoveries of madecassoside, madecassic acid and IS from spiked plasma samples were > 81%. The developed method was successfully applied to the pharmacokinetic study of madecassoside and madecassic acid in rats after an oral administration of madecassoside. During initial 7 days of dosing, the cmax and AUC of madecassoside were greatly decreased and Vd/F was markedly increased in CIA rats, and no significant difference was observed on the first day of dosing. In contrast, the T1/2, cmax and AUC of madecassic acid were significantly increased, and Ke of madecassic acid was greatly decreased in CIA rats compared with normal rats. Along with repeated administration of madecassoside, the differences of pharmacokinetic parameters of both madecassoside and madecassic acid between CIA and normal rats gradually subsided. The pharmacokinetic characteristics of both madecassoside and madecassic acid in rats were significantly altered by arthritis status, and the differences of pharmacokinetic parameters between arthritis and normal rats coincide with the severity of arthritis.

Formulation, standardization and pharmacological studies of Saraswataristam: a polyhedral preparation

Ayurvedic preparations achieved paramount importance in contemporary life owing to the safety and efficacy when compared with those of synthetic drugs. But due to lack of proper standardization at each stage from starting to culmination results in inferior quality and less demand. Saraswataristam, a fermented ayurvedic preparation, has been employed for treating central nervous system disorders and dermatological problems. Saraswataristam, containing Centella asiatica [L] urban [Umbelliferae] as the major ingredient is prepared as per the Ayurvedic Pharmacopoeial specifications, characterized and standardized for determining the quality, safety and efficacy of herbs used in it. Saraswataristam was prepared and subjected to phytochemical screening by FTIR analysis and HPTLC fingerprinting, heavy metal determination by AAS, determination of alcohol content, test for E. coli, S. aureus, aerobic bacteria, yeasts and mould, oral toxicity studies and anti-epileptic activity by MES method. The physico-chemical studies showed total ash content as 1.1%, extractive values and some trace elements such as Lead, Mercury, Cadmium and Arsenic with 3.1, 0.047, 0.17 and 0.46 ppm respectively and all values are found within the acceptable limits specified by WHO. FTIR and HPTLC studies showed the presence of asiaticoside in Saraswataristam, resulting in its chemical standardization. The formulation showed signs of dose dependent significant [P<0.001] reduction in various episodes of epileptic seizures in comparison with standard phenytoin, thereby making it biologically standardized. The physico-chemical and pharmacological analysis to standardize Saraswataristam confirmed its use as a safe anti-epileptic ayurvedic formulation.

Simultaneous determination of madecassoside, asiaticoside and their aglycones in Centella asiatica (L.) urban extracts by RP-HPLC / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a HPLC method for simultaneous determination of four major constituents (madecassoside, asiaticoside, madecassic acid and asiatic acid) in Centella asiatica (L.) urban extracts.</p><p><b>METHODS</b>The analysis was performed on an Agilent 1100 HPLC system with a ZORBAX Eclipse XDB-C8 column (4.6 mm×150 mm, 5μm). The four major constituents were separated with gradient mobile phase that consists of 1mmol/L potassium dihydrogen phosphate and acetonitrile at the detection wavelength of 205 nm.</p><p><b>RESULTS</b>The four major constituents all had good linear response in the determination ranges (R(2)≥0.9998). The average recoveries (n=9) were 97.4%, 93.7%, 97.5% and 99.8% with RSDs of 3.4%, 1.4%, 4.7% and 4.4%, respectively.</p><p><b>CONCLUSION</b>The developed method is sensitive and has good reproducibility, which can be used as a reference for quality control of Centella asiatica (L.) urban extracts.</p>

Attenuation of collagen induced arthritis by Centella asiatica methanol fraction via modulation of cytokines and oxidative stress / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the anti-inflammatory, antioxidant and anti-arthritic effects of Centella asiatica methanolfraction (CaME) on collagen-induced arthritis (CIA), an animal model of rheumatoid arthritis.</p><p><b>METHODS</b>Arthritis was induced in female wistar rats by immunization with porcine type II collagen. The CIA rats were treated orally with CaME (50, 150, and 250 mg/kg/day) for 15 d (beginning on day 21 of the experimental period). The clinical, histological, biochemical, and immunological parameters were assessed.</p><p><b>RESULTS</b>CaME treatment (150 and 250 mg/kg) significantly attenuated the severity of CIA and reduced the synovial inflammation, cartilage erosion, and bone erosion as evident from both histological and radiographic data. The escalated plasma levels of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-12 alongwith nitric oxide in CIA rats decreased significantly on CaME treatment. The serum levels of type-II collagen antibody were significantly lower in rats of CaME (150 and 250 mg/kg) treated group than those in the arthritic group. Furthermore, by inhibiting the above mediators, CaME also contributed towards the reversal of the disturbed antioxidant levels and peroxidative damage.</p><p><b>CONCLUSION</b>Our results clearly indicate that oral administration of CaME suppresses joint inflammation, cytokine expression as well as antioxidant imbalance, thereby contributing to an amelioration of arthritis severity in CIA rats.</p>

Anti-diabetic effect of a combination of andrographolide-enriched extract of Andrographis paniculata (Burm f.) Nees and asiaticoside-enriched extract of Centella asiatica L. in high fructose-fat fed rats.

Traditionally, a combination of medicinal plants is commonly used for lowering blood glucose in diabetic patients in order to provide additional benefits of the single drug. A. paniculata and C. asiatica are two traditional medicines form South Asian and Southeast Asain countries consumed by people for treating daibates mellitus and its complications. Hyperglycemia in the rats was stimulated by high fructose-fat diet that consists of 36% fructose, 15% lard, and 5% egg yolks in 0.36 g/200 g body weight for 70 days. The rats were orally administered with the combination of andrographolide-enriched extract of A. paniculata (AEEAP) leaves and asiaticoside-enriched extract of C. asiatica (AEECA) herbs from day 70 for 7 days. Antidiabetic activity was evaluated by estimating mainly the blood glucose levels and other parameters such as HDL, LDL, cholesterol and triglyceride. The results showed that combination at the ratio of 70:30 exhibited a promosing antidiabetic effect in high-fat-fructose-fed rat, and exhibited sinergistic effects on blood cholesterol and HDL levels. It can be concluded that its antidiabetic effect was better than that of single treatment of AEEAP or AEECA. That combination was also potential to develop as a blood glucose-lowering agent for diabetic patients.

Anti-proliferative activities of centella asiatica extracts on human respiratory epithelial cells in vitro / Actividades antiproliferativas del extracto de centella asiatica sobre células epiteliales respiratorias humanas in vitro

Centella asiatica or "pegaga" is well known for its ability in promoting wound healing. This study focused on the effect of Centella asiatica on the proliferation of human respiratory epithelial (RE) cells. RE cells were cultured using co-culture techniques until first passage (P1). Viability cell test by tryphan blue dye exclusion assay showed that there was high percentage of cell viability at both P0 (74 percent) and P1 (91.61 percent). Triplicate MTT assays were carried out with different concentrations of C. asiatica from 15.6 ppm, 31.3 ppm, 62.5 ppm, 125 ppm, 250 ppm, 500 ppm, 1000 ppm, until 2000 ppm. The higher the concentration of C. asiatica, the more inhibitory effect was seen. C. asiatica aqueous extract at concentration 1000 ppm and 2000 ppm demonstrated a significant (p<0.05) inhibitory effect on human RE cells proliferation on day 4 and day 7 after treatment. This provides potential use of C. asiatica extract for the treatment of conditions with respiratory epithelial cells overgrowth.

La Centella asiatica o "pegaga" es conocida por su capacidad para promover la cicatrización de heridas. Este estudio se centró en el efecto de la Centella asiatica sobre la proliferación de las células del epitelio respiratorio (ER) humano. Las células del ER se cultivaron usando técnicas de co-cultivo hasta el primer paso (P1). La prueba la viabilidad celular mediante el ensayo de exclusión del colorante azul de tripano demostró un alto porcentaje de viabilidad celular tanto en P0 (74 por ciento) y P1 (91,61 por ciento). Ensayos de MTT por triplicado se llevaron a cabo con diferentes concentraciones de C. asiatica, desde 15,6 ppm, 31,3 ppm, 62,5 ppm, 125 ppm, 250 ppm, 500 ppm, 1000 ppm, hasta 2000 ppm. Con una mayor concentración de C. asiatica, se observó un mayor efecto inhibitorio. El extracto acuoso de C. asiatica a una concentración de 1000 ppm y 2000 ppm demostró un efecto inhibidor significativo (p<0,05) sobre la proliferación de células del ER humano los días 4 y 7 posterior al tratamiento. Esto proporciona el uso potencial del extracto de C. asiatica para el tratamiento de condiciones con crecimiento excesivo células epiteliales respiratorias.