Titrated Extract Of Centella Asiatica Research Articles

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

PurposeIn 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.MethodsAn 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).ResultsPA-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.ConclusionsOur results demonstrate that the combination of TECA and AST could be a promising therapeutic agent for AD by inhibiting NF-κB signaling.

Titrated extract of Centella asiatica increases hair inductive property through inhibition of STAT signaling pathway in three-dimensional spheroid cultured human dermal papilla cells.

Dermal papilla (DP) is a pivotal part of hair follicle, and the smaller size of the DP is related with the hair loss. In this study, we investigated the effect of titrated extract of Centella asiatica (TECA) on hair growth inductive property on 3D spheroid cultured human DP cells (HDP cells). Significantly increased effect of TECA on cell viability was only shown in 3D sphered HPD cells, not in 2D cultured HDP cells. Also, TECA treatment increased the sphere size of HDP cells. The luciferase activity of STAT reporter genes and the expression of STAT-targeted genes, SOCS1 and SOCS3, were significantly decreased. Also, TECA treatment increased the expression of the hair growth-related signature genes in 3D sphered HDP cells. Furthermore, TECA led to downregulation of the level of phosphorylated STAT proteins in 3D sphered HDP cells. Overall, TECA activates the potential of hair inductive capacity in HDP cells.

Anti-Inflammatory Effect of Titrated Extract of Centella asiatica in Phthalic Anhydride-Induced Allergic Dermatitis Animal Model.

Centella asiatica has potent antioxidant and anti-inflammatory properties. However, its anti-dermatitic effect has not yet been reported. In this study, we investigated the anti-dermatitic effects of titrated extract of Centella asiatica (TECA) in a phthalic anhydride (PA)-induced atopic dermatitis (AD) animal model as well as in vitro model. An AD-like lesion was induced by the topical application of five percent PA to the dorsal skin or ear of Hos:HR-1 mouse. After AD induction, 100 μL of 0.2% and 0.4% of TECA (40 μg or 80 μg/cm2) was spread on the dorsum of the ear or back skin three times a week for four weeks. We evaluated dermatitis severity, histopathological changes and changes in protein expression by Western blotting for inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and NF-κB activity, which were determined by electromobility shift assay (EMSA). We also measured TNF-α, IL-1β, IL-6, and IgE concentration in the blood of AD mice by enzyme-linked immunosorbent assay (ELISA). TECA treatment attenuated the development of PA-induced atopic dermatitis. Histological analysis showed that TECA inhibited hyperkeratosis, mast cells and infiltration of inflammatory cells. TECA treatment inhibited expression of iNOS and COX-2, and NF-κB activity as well as the release of TNF-α, IL-1β, IL-6, and IgE. In addition, TECA (1, 2, 5 μg/mL) potently inhibited Lipopolysaccharide (LPS) (1 μg/mL)-induced NO production, expression of iNOS and COX-2, and NF-κB DNA binding activities in RAW264.7 macrophage cells. Our data demonstrated that TECA could be a promising agent for AD by inhibition of NF-κB signaling.

Asiatic acid inhibits pulmonary inflammation induced by cigarette smoke

Asiatic acid (AA) is one of the major components of Titrated extract of Centella asiatica (TECA), which has been reported to possess antioxidant and anti-inflammatory activities. The purpose of this study was to investigate the protective effect of AA on pulmonary inflammation induced by cigarette smoke (CS). AA significantly attenuated the infiltration of inflammatory cells in bronchoalveolar lavage fluid (BALF) of CS exposure mice. AA also decreased ROS production and NE activity, and inhibited the release of proinflammatory cytokines in BALF. AA reduced the recruitment of inflammatory cells and MCP-1 expression in lung tissue of CS exposure mice. AA also attenuated mucus overproduction, and decreased the activation of MAPKs and NF-kB in lung tissue. Furthermore, AA increased HO-1 expression and inhibited the reduced expression of SOD3 in lung tissue. These findings indicate that AA effectively inhibits pulmonary inflammatory response, which is an important process in the development of chronic obstructive pulmonary disease (COPD) via suppression of inflammatory mediators and induction of HO-1. Therefore, we suggest that AA has the potential to treat inflammatory disease such as COPD.

센텔라 아시아티카 정량추출물을 함유한 Water in Oil in Water (W<sub>1</sub>/O/W<sub>2</sub>) 에멀젼 제조에 관한 연구

Titrated extract of Centella asiatica (TECA), which is poorly soluble in water is well known for wound healing and anti wrinkle agent. This study was conducted to find the optimum condition for the preparation of water in oil in water (W1/O/W2) emulsion containing TECA. Solubility of TECA were measured by UV spectrophotometer. 2.55 g of TECA was dissolved in solution composed of dipropylene glycol (40.0 g), ethanol (20.0 g), and water (10.0 g). Factors affecting stability of the emulsions (W1/O, W1/O/W2) was investigated. The optimum conditions for the preparation of W1/O emulsion was composed of dipropylene glycol : ethanol : water : TECA in a weight ratio of 40.0 : 20.0 : 10.0 : 2.5 for water phase and squalane : cetyl PEG/PPG-10/1 dimethicone : cetearyl alcohol in a weight ratio of 22.5 : 4.0 : 2.5 for oil phase. The optimum conditions for the preparation of W1/O/W2 multiple emulsion was composed of water : W1/O emulsion : polysorbate 80 : carbomer : triethanolamine in a weight ratio of 55.8 : 40.0 : 4.0 : 0.1 : 0.1.

Asiaticoside, a component of Centella asiatica, inhibits melanogenesis in B16F10 mouse melanoma.

Melanogenesis is the process of generating pigmentation via melanin synthesis and delivery. Three key enzymes, tyrosinase, tyrosinase-related protein 1 (TRP1) and TRP2, metabolize melanin from L-tyrosine. Melanin synthesizing enzymes are regulated by microphthalmia-associated transcription factor (MITF). The titrated extract of Centella asiatica (TECA) contains the major components asiatic acid, asiaticoside and madecassic acid. The present study revealed that TECA reduces the melanin content in melanocytes. Moreover, the asiaticoside contained in TECA modulated melanogenesis by inhibiting tyrosinase mRNA expression. The decrease in tyrosinase mRNA levels was mediated through MITF. Uniquely, asiaticoside inhibited MITF by decreasing its DNA binding affinity. In conclusion, the results of the present study indicate that asiaticoside treatment may have beneficial effects in hyperpigmentation diseases or for skin whitening.

Titrated extract of Centella asiatica provides a UVB protective effect by altering microRNA expression profiles in human dermal fibroblasts

The titrated extract of Centella asiatica (TECA) is a reconstituted mixture comprising of asiatic acid, madecassic acid, asiaticoside and madecassoside, and is used as a therapeutic agent in wound healing and also as an anti-microbial, anticancer and anti-aging agent. Although these properties and the associated cell signaling pathways have been elucidated, the cellular mechanism of anti-photoaging upon ultraviolet (UV) exposure in normal human dermal fibroblasts (NHDFs) remains unknown. In this study, we investigated the photoprotective role of TECA via microRNA (miRNA) expression profiling analysis. Low dose of TECA did not exhibit toxicity and showed a protective effect against UVB irradiation in NDHFs. miRNA microarray experiments revealed that specific miRNAs were altered by TECA stimulation in UVB-irradiated NHDFs. Functional bioinformatic analysis showed that the putative target genes of the altered miRNAs were associated with the positive regulation of cell proliferation, anti-apoptosis, small GTPase- and Ras-mediated signal transduction and activation of MAPKK. Therefore, these results suggest that TECA may serve as a potential natural chemoprotective agent against UVB-mediated damage in NHDFs through changes in the expression of specific miRNAs.

Evaluation of the wound healing potentials of two subspecies of Hypericum perforatum on cultured NIH3T3 fibroblasts

For centuries, Hypericum perforatum has been used in folk medicine to treat wounds. In the present study, the wound healing activities of extracts of H. perforatum ssp. perforatum (HPP) and H. perforatum ssp. veronense (HPV) were evaluated by comparing with a titrated extract of Centella asiatica (TECA) on NIH3T3 fibroblasts. The cells were incubated with the extracts. Using microscopical methods by staining cells, mitotic ability, morphological changes and collagen production in the fibroblasts were evaluated as parameters to approach possible wound repair mechanism(s). The wound healing activity caused an increase in the percentage of polygonal fibroblasts and in collagen granule numbers in fibroblasts of HPP and HPV. At 1 µg/mL, HPP caused a greater increase in mitotic cell numbers than HPV. The most increases in polygonal cell numbers were observed with HPV at 1 and 10 µg/mL. The number of collagen granules was increased with 1 µg/mL HPV being higher than HPP. As an indication of cytotoxic effects, round cells in response to HPV and HPP extracts, were found to be increased in concentrations higher than 10-50 µg/mL. The results indicated that the two subspecies of H. perforatum have different wound healing profiles as a result of the fibroblast migration and stimulation of collagen synthesis.

Wound-healing activity of St. John's Wort ( Hypericum perforatum L.) on chicken embryonic fibroblasts

Wound-healing effect of St. John's Wort ( Hypericum perforatum L.) extract was evaluated by comparing with dexpanthenol and titrated extract of Centella asiatica (TECA) on cultured chicken embryonic fibroblasts. Chicken embryonic fibroblasts from fertilized eggs were incubated with the plant extract, dexpanthenol and TECA. Using microscopical methods by staining cells, mitotic ability, morphologic changes and collagen production in the cultured fibroblasts were evaluated as parameters to approach its mechanism of action in wound repair. Findings obtained in the present study indicated that Hypericum perforatum extract exhibited a wound-healing activity whose mechanism of action is similar to that of TECA. Wound-healing activity of Hypericum perforatum extract seems to be mainly due to the increase in the stimulation of fibroblast collagen production and the activation of fibroblast cells in polygonal shape, which plays a role in wound repair by closing damaged area. The findings demonstrated the wound-healing activity of Hypericum perforatum, which has previously been based on ethnomedical data.

Advanced formulation and pharmacological activity of hydrogel of the titrated extract ofC. asiatics

Titrated extract of Centella asiatica (TECA) contains three principal ingredients, asiaticoside (AS), asiatic acid (AA), and madecassic acid (MA). These components are known to be clinically effective on systemic scleroderma, abnormal scar formation, and keloids. However, one problem associated with administration of TECA is its low solubility in aqueous as well as oil medium. In this study, various nonionic surfactants and bile salts as anionic surfactant were tested and screened for solubilizing TECA with a view to developing topical hydrogel type of ointment which is stable physicochemically, and has better pharmacological effects. When TECA was incorporated into various nonionic surfactant systems, labrasol had the most potent capacity for solubilizing TECA. In cases of bile salt systems, Na-deoxycholate (Na-DOC) had foremost solubilizing capacity, even more than labrasol. In differential scanning calorimetric study, the peaks of AA, MA, AS and Na-DOC disappeared at the coprecipitate of 1% TECA and 1% Na-DOC, suggesting the optimum condition of Na-DOC for solubilizing TECA. When the physicochemical stability of hydrogel containing this mixture was assessed, it was stable at room temperature for at least one month. Pharmacologically it significantly decreased the size of wound area at the 9th day when applied to the wound area of rat dorsal skin. Taken together, solubility of TECA was dramatically improved by using nonionic and anionic surfactant systems, and Na-DOC was found to be the most effective solubilizer of TECA in formulating a TECA-containing hydrogel typed ointment. Moreover this gel was considered to be applicable to clinical use for wound healing effect.

니오좀을 이용한 병풀 추출물 외용제의 제조 및 평가

For preventing and curing the stretching mark, TECA Niosome/W/O system creams were formulated using Titrated Extract of Centella Asiatica (TECA) which is well known for its excellent wound healing effect. The lipid-water partition coefficients and the stabilities of TECA were evaluated and TECA Niosome/W/O system (TECA N/W/O) creams were prepared with different concentrations of cetyl alcohol and ceramide. TECA N/W/O cream was evaluated with respect to their rheological properties, permeation through excised skin of hairless mouse and in vitro and in vivo accumulation in the skin of hairless mouse. In addition, dermal thicknesses of hairless mouse skins were determined following the in vivo application of TECA N/W/O cream and control cream. TECA N/W/O creams showed pseudoplastic flow and hysteresis loop. The permeation of TECA from formulations through excised skin of hairless mouse did not observed. Amount of accumulated drug in the excised skin of hairless mouse was deσeased with an increase in the concentration of cetyl alcohol and showed no relationship with concentration of ceramide. Amount of accumulated drug in formulation A-3 was higher than in niosome suspension and other formulations. In in vivo experiment, amount of accumulated drug in formulation A-2 and A-3 was much higher than that of niosome suspension. Being treated with the N/W/O cream for 8 weeks, the dermal thickness of hairless mouse skin was increased 3.2 times than that of 16 weeks-control group.

Development of a novel dosage form for intramuscular injection of titrated extract of Centella asiatica in a mixed micellar system

Titrated extract of Centella asiatica (TECA), a drug used in treating systemic scleroderma, is poorly water-soluble. A conventional dosage form for the intramuscular injection of TECA, propylene glycol (PG)-based TECA solution, causes severe pain after intramuscular injection. To improve the solubility of TECA and reduce pain after injection, mixed micellar systems composed of 10% surfactant mixture (Tween 20 and Tween 85) and 90% phosphate-buffered saline, pH 7.0 (PBS) were prepared. As the ratio of Tween 20 to Tween 85 increased from 0:10 to 10:0, the solubility of TECA in the mixed micellar systems increased from 7- to 26-fold compared to that in PBS (pH 7.0). The droplet size of micelles gradually decreased with the increasing ratio of Tween 20 to Tween 85 from 0:10 to 4:6, followed by an abrupt decrease in size above the ratio of 6:4. Furthermore, the micellar systems prepared with Tween 20 and Tween 85 at the ratio of 6:4, 8:2 or 10:0 could solubilize TECA more than 10 mg/ml and the resultant droplet sizes were less than 2 μm. No significant changes were observed in the droplet sizes and asiaticoside contents in these micellar formulations during storage, indicating these systems are stable for at least 60 days. Their osmotic pressures were remarkably lower than those of PG-based TECA solution and similar to that of saline solution, irrespective of dilution ratios. Most importantly, they markedly reduced the number of writhes compared with PG-based TECA solution after injection to mice. All of these results suggest that these three TECA micellar formulations prepared with Tween 20 and Tween 85 improved the solubility of TECA and reduced pain following injection, possibly due to the decrease in osmotic pressure. Thus, these micellar formulations composed of optimum ratios of Tween 20 and Tween 85 may have a potential as dosage forms for the intramuscular injection of a poorly water-soluble TECA.

Preparation and evaluation of a titrated extract of Centella asiatica injection in the form of an extemporaneous micellar solution

The micellar solution of titrated extract of Centella asiatica (TECA) was formulated by solubilizing TECA into a mixture of propylene glycol and ethoxylated hydrogenated castor oil. An extemporaneous TECA micelle was then successfully prepared by diluting this original micellar formulation with water, saline or isotonic glucose solution. A mixture of propylene glycol and ethoxylated hydrogenated castor oil achieved an acceptable solubilization of TECA up to 180 mg/ml via a formulation of micelle. The drug content of the original micellar formulation stored at 55°C over 60 days was almost constant. The extemporaneous TECA micelle prepared by diluting the original micellar formulation was physically stable for up to 23 h with shaking and 55 h without shaking depending on dilution ratio and medium. The estimated distribution of mean particle size was between 15.9 and 32.6 nm. The osmotic pressure and erythrocytic hemolysis were measured for the formulation with various media. The osmotic pressure of the formulation was found to be much lower than that in a commercially formulated propylene glycol-based injection. The erythrocytic hemolysis by the micellar solution was much more reduced compared with that by the propylene glycol-based preparation. Pain score after the peritoneal injection of the micellar solution was assessed by counting the number of writhe in ICR mice and compared with that in the conventional injection. The injection of extemporaneous TECA micellar solution did not show any single writhe in mice as like as saline. This indicates that pain is reduced dramatically. These results indicated that a micellar solubilization, followed by an extemporaneous dilution, might be a choice of formulation for TECA injection with reducing the pain arising from the intramuscular injection.