Concentrations Of Asiaticoside Research Articles

Application of machine learning models for green andsimultaneous determination of asiaticoside andmadecassoside in Centella asiatica

The artificial neuron network (ANN), which is a subfield of machine learning, has been widely applied in analytical chemistry for classification/pattern recognition, prediction, and modeling. ANN combined with UV spectrometry can be used to tackle the problem of overlapping spectra of a complicated matrix of herbal medicine. In the present study, ANN has been used for simultaneous determination of asiaticoside andmadecassoside in Centella asiatica collected from various provinces in Viet Nam based on UV spectra of standard reference and spiked samples. The absorption spectra of 108 C. asiatica samples were recorded in (190-250) nm of wavelength with an interval of 1 nm (61 variables) were used for data acquisition. An ANN model using traincgb function with 40 neural hidden layers was trained. The correlation coefficients were all higher than 0.9999. The concentration of asiaticoside and madecassoside in all tested samples revealed a good recovery, as compared to the referenced values. TheANN model can be considered as effective, time-saving and environment-friendly quantitative analysis tool for herbal medicine ® 2024 Journal of Science and Technology - NTTU

Examining the Effects of Asiaticoside on Dental Pulp Stem Cell Viability and Proliferation: A Promising Approach to Root Canal Treatment.

This study aims to evaluate the impact of asiaticoside (AC) on the viability and proliferation of dental pulp stem cells (DPSCs), considering the known negative effects of routinely used intracanal medicaments. This evaluation will be compared with the outcomes from using traditional intracanal medicaments, specifically triple antibiotic paste (TAP) and calcium hydroxide [Ca(OH)2]. The DPSCs were obtained from the third molars of an adult donor. The application of flow cytometry was employed to do a phenotypic analysis on DPSCs using CD90, CD73, CD105, CD34, CD14, and CD45 antibodies. The methylthiazol tetrazolium (MTT) assay was employed to assess cellular viability. The cells were treated with different concentrations of TAP and Ca(OH)2 (5, 2.5, 1, 0.5, and 0.25 mg/mL), along with AC (100, 50, 25, 12.5, and 6.25 µM). A cell proliferation rate was performed at 3, 5, and 7 days. The characterization of DPSCs was conducted by flow cytometry analysis, which verified the presence of mesenchymal cell surface antigen molecules (CD105, CD73, and CD90) and demonstrated the absence of hematopoietic markers (CD34, CD45, and CD14). Cells treated with concentrations over 0.5 mg/mL of TAP and Ca(OH)2 showed a notable reduction in cell viability in comparison to the untreated cells (p < 0.05). Additionally, the cells treated with different concentrations of AC 12.5, 6.25, 25, and 50 µM did not differ significantly from the untreated cells (p > 0.05). Nevertheless, cells treated with concentrations of 100 µM showed a significant reduction in viability compared to the untreated cells (p < 0.05). After a period of 7 days, it was noted that cells exposed to three different concentrations of AC (50, 25, and 12.5 µM) had a notable rise in cell density in comparison to TAP and Ca(OH)2 (p < 0.05). Furthermore, cells that were exposed to a concentration of 12.5 µM exhibited the highest cell density. The cellular viability of the AC-treated cells was superior to that of the TAP and Ca(OH)2-treated cells. Moreover, the AC with a concentration of 12.5 µM had the highest degree of proliferation. This study underscores the importance of evaluating alternative root canal medicaments and their effects on DPSCs' growth and vitality. The findings on AC, particularly its influence on the survival and proliferation of DPSCs, offer valuable insights for its probable use as an intracanal medication. This research contributes to the ongoing efforts to identify safer and more effective intracanal treatments, which are crucial for enhancing patient outcomes in endodontic procedures. How to cite this article: Alazemi MJ, Badawi MF, Elbeltagy MG, et al. Examining the Effects of Asiaticoside on Dental Pulp Stem Cell Viability and Proliferation: A Promising Approach to Root Canal Treatment. J Contemp Dent Pract 2024;25(2):118-127.

Heterogeneous ensemble machine learning to predict the asiaticoside concentration in centella asiatica urban

This study proposes a novel heterogeneous ensemble machine learning methodology to predict the concentration of asiaticoside in Centella asiatica (CA-CA) in the context of the lack of an effective prediction method capable of accurately estimating its quantity based on various growing environmental factors. The accurate prediction of the asi-aticoside concentration in CA-CA holds great significance in optimizing cultivation practices and improving the efficacy of the derived medicinal products. The presented approach aims to address this crucial need by employing a diverse ensemble of machine learning techniques. The proposed model integrates several machine learning tech-niques, including the standard long short-term memory (LSTM), gated recurrent unit (GRU), convolutional long short-term memory (ConvLSTM), and attention-based LSTM, by utilizing a differential evolution algorithm to optimize the ensemble model's weights. The developed model is called the heterogeneous ensemble machine learning model (He-ML). Experimental results demonstrate that the He-ML achieves an im-pressive root-mean-square error (RMSE) value of 4.76, which is up to 12.48 % lower than the RMSE. The findings highlight the advantages of employing an ensemble model over a single model, as the ensemble model achieves an RMSE value that is 14.67 % lower than that of the individual machine learning model. The utilization of differential evolution as the decision fusion strategy provides a notable improvement over the unweighted average approach. As a result, the RMSE value achieved is 8.46 % lower than that obtained with the unweighted average (UWA) technique.

Natural drug asiaticoside inhibits osteoclast differentiation and promotes repair of steroid-induced osteonecrosis of femoral head via RANKL pathway

Asiaticoside is one of the most classic traditional Chinese medicines. The interaction between Receptor activator of nuclear factor κB (RANK) and Receptor Activator of Nuclear Factor-κ B Ligand (RANKL) activates a series of intracellular signaling pathways. Our research explored the mechanism of asiaticoside inhibiting osteoclast differentiation and repair of steroid-induced osteonecrosis of femoral head (SIONFH) through RANKL pathway. Asiaticoside’s effect on osteoclasts was determined by Microneedle Therapy System (MTS) method, and the number of Tartrate resistant acid phosphatase (TRAcP) multinucleated cells (3 nuclei) was quantitatively analyzed. Under different dose treatments, the changes of bone function were detected by eroded area and TRAcP staining on hydroxyapatite-coated plate, and the expression of different osteocyte formation and osteogenic differentiation were detected. Asiaticoside weakened the osteoclast function induced by RANKL. When the concentration of asiaticoside was 10 μmol/L, osteoclast formation regulatory genes (NFATc1 and Acp5) and osteoclast cell function-related genes (AP-1 and P65) were significantly downregulated in the presence of RANKL. Asiaticoside (10 μmol/L) significantly decreased calcium oscillations induced by RANKL via preventing RANKL-mediated NF-κB activation and Ca2+ oscillation.

&lt;i&gt;In Vitro&lt;/i&gt; Wound Healing Effect of Asiaticoside Extracted from &lt;i&gt;Centella asiatica&lt;/i&gt; (‘Pegaga’) on Human Gingival Fibroblast Cell Line

Asiaticoside is a bioactive compound found in the traditional plant Centella asiatica (Asiatic pennywort or ‘Pegaga’) generally used for wound healing applications. Numerous studies have discussed the potential benefits of asiaticoside on different human cells such as keratinocytes and dermal fibroblast cells in healing of wounds. However only very few studies have been conducted to investigate its healing effect on cells originated from human oral cavity. The present study aimed to determine the potential of asiaticoside on human gingival fibroblast cells. Cytotoxic activities of the compounds were assessed by MTT assay. The wound healing was examined by scratch assay. The effect of asiaticoside on Col1A1 gene expression was also analyzed using qRT-PCR. Col1A1 is known to play a crucial role in wound healing. The MTT assay result showed that the maximum tolerable concentration of asiaticoside was 0.25 mg/ml. The scratch assay revealed that asiaticoside significantly accelerated the wound healing compared to the negative control (P&lt;0.05). Moreover, the qRT-PCR demonstrated that asiaticoside markedly increased Col1A1 mRNA expression. These results proved asiaticoside as a potential candidate for wound healing agent in dentistry.

Chemotaxonomic Studies on Centella asiatica (L.) Urb. from Varied Phytogeographical Conditions of India for Its Industrial Prospection

Centella asiatica (L.) Urb. (Apiaceae) with high commercial value is an important herb in traditional Indian medicine. In the present study, comparative chemoprofiling was performed on various samples of C. asiatica (CA) collected from different phytogeographical zones of India, using high-performance thin-layer chromatography. Three biologically active phytochemicals, viz. asiaticosides, madeccasosides and asiatic acid, were investigated in 35 samples of CA. Variation in concentrations of asiaticoside, madeccasoside and asiatic acid was observed as 0.03–0.66%, 0.007–2.688% and 0.006–0.44%, respectively. The mobile phase which was used for asiaticoside and madecassoside was butanol:ethyl acetate:water, and for asiatic acid, it was toluene:ethyl acetate:formic acid. Among all samples, significant variation was observed in asiaticoside and madeccasoside from Northern Himalayas, while very small variation was found in asiatic acid from Deccan region. The results of the present study are of industrial importance in selection of phytogeographical regions of India for collection and to get maximum contents of active targeted phytochemicals.

Asiaticoside induces osteogenic differentiation of human periodontal ligament cells through the Wnt pathway.

Asiaticoside is a compound isolated from Herb Centella asiatica, which has been shown to promote osteogenic differentiation of human periodontal ligament (hPDL) cells. This study investigated the molecular mechanism underlying the asiaticoside-induced osteogenic differentiation of hPDL cells. hPDL cells were incubated with various concentrations of asiaticoside to test cell viability by MTT assay. The mRNA expression levels were analyzed by using quantitative real-time polymerase chain reaction (PCR). Osteogenic differentiation was determined by alkaline phosphatase activity assay and alizarin red staining. The subcellular localization of β-catenin was analyzed by both immunofluorescence and western blot. The results showed that asiaticoside had no effect on the cell viability at any of the tested concentrations. Real-time PCR revealed that osterix (OSX) and dentin matrix protein1 (DMP1) mRNA were significantly enhanced by asiaticoside treatment. Alkaline phosphatase activity and in vitro mineralization were also significantly induced. Interestingly, asiaticoside dose-dependently increased WNT3A mRNA expression, but not WNT5A and WNT10B. The activation of Wnt signaling was shown to result in nuclear accumulation of β-catenin as evaluated by immunofluorescence staining and western blot analysis. Pre-treatment with recombinant human Dickkopf1 (rhDKK1) inhibited asiaticoside-induced β-catenin nuclear translocation and osteoblast marker gene expression. Moreover, rhDKK1 attenuated asiaticoside-induced DMP1 protein expression. The data demonstrate that asiaticoside induces osteogenic differentiation of hPDL cells by activating the Wnt/β-catenin signaling pathway. The findings suggest that asiaticoside could be used as a novel therapeutic drug for periodontal tissue regeneration.

Asiaticoside increases aquaporin-3 protein expression in the cytoplasm of normal human epidermal keratinocytes

&lt;p&gt;Introduction&lt;br /&gt;Skin hydration decreases with aging. Aquaporin-3 (AQP3) is a major protein that plays a role in skin hydration, therefore it is a novel target for skin moisturizing treatment. Retinoic acid (RA) as a well-known active agent in antiaging treatment increases AQP3 expression, but frequently causes harmful side effects. Asiaticoside, a saponin compound isolated from Centella asiatica (CA) is also known as an antiaging cosmetic and plays a role in wound healing. The aim of this study was to evaluate and compare the effect of asiaticoside isolated from CA and the effect of RA on the AQP3 expression in normal human epidermal keratinocytes (NHEKs).&lt;/p&gt;&lt;p&gt;Methods &lt;br /&gt;An experimental laboratory study was performed using primary NHEKs that were derived from the foreskin of a boy. AQP3 expression in NHEKs was examined in vitro after the cells were incubated for 24 hours with asiaticoside or with RA at several concentrations. The AQP3 expression was evaluated by immunocytochemistry and quantitatively analyzed by Image-J software. Independent t-test and one-way ANOVA were used to analyze the data, followed by post-hoc Tukey test.&lt;/p&gt;&lt;p&gt;Results&lt;br /&gt;There was an increasing trend of AQP3 expression upon exposure to asiaticoside at all concentrations compared to the control group. However, RA exposure seemed to induce a higher level of AQP3 expression. Asiaticoside effected a lower increase in AQP3 expression in NHEKs than did RA (p=0.042). Optimal results were achieved at 1 mg/mL concentration of asiaticoside.&lt;/p&gt;&lt;p&gt;Conclusions&lt;br /&gt;Asiaticoside isolated from CA can enhance the AQP3 expression in NHEKs. Therefore it can be used as an active ingredient in cosmetic moisturizer formulation for dry skin treatment.&lt;/p&gt;

Combination of three laboratory data as predictor of severe dengue in adults : a retrospective cohort study

&lt;p&gt;Introduction&lt;br /&gt;Skin hydration decreases with aging. Aquaporin-3 (AQP3) is a major protein that plays a role in skin hydration, therefore it is a novel target for skin moisturizing treatment. Retinoic acid (RA) as a well-known active agent in antiaging treatment increases AQP3 expression, but frequently causes harmful side effects. Asiaticoside, a saponin compound isolated from Centella asiatica (CA) is also known as an antiaging cosmetic and plays a role in wound healing. The aim of this study was to evaluate and compare the effect of asiaticoside isolated from CA and the effect of RA on the AQP3 expression in normal human epidermal keratinocytes (NHEKs).&lt;/p&gt;&lt;p&gt;Methods &lt;br /&gt;An experimental laboratory study was performed using primary NHEKs that were derived from the foreskin of a boy. AQP3 expression in NHEKs was examined in vitro after the cells were incubated for 24 hours with asiaticoside or with RA at several concentrations. The AQP3 expression was evaluated by immunocytochemistry and quantitatively analyzed by Image-J software. Independent t-test and one-way ANOVA were used to analyze the data, followed by post-hoc Tukey test.&lt;/p&gt;&lt;p&gt;Results&lt;br /&gt;There was an increasing trend of AQP3 expression upon exposure to asiaticoside at all concentrations compared to the control group. However, RA exposure seemed to induce a higher level of AQP3 expression. Asiaticoside effected a lower increase in AQP3 expression in NHEKs than did RA (p=0.042). Optimal results were achieved at 1 mg/ml concentration of asiaticoside.&lt;/p&gt;&lt;p&gt;Conclusions&lt;br /&gt;Asiaticoside isolated from CA can enhance the AQP3 expression in NHEKs. Therefore it can be used as an active ingredient in cosmetic moisturizer formulation for dry skin treatment.&lt;/p&gt;

Multiple Oral Dosing Pharmacokinetics of Standardized Extract of Centella asiatica ECa 233 and Its Inductive Effect on Efflux Transporters in Rats

AbstractECa 233 is a standardized extract of Centella asiatica, characterized as a white powder containing triterpenoid glycosides not less than 80% with a ratio of madecassoside to asiaticoside of 1.5±0.5:1. Although pharmacological and toxicological profiles of ECa 233 have been successively reported, the pharmacokinetic data needed for further therapeutic development are not fully elucidated. This study aimed to investigate the pharmacokinetics of multiple oral dosing of ECa 233 at 100 mg/kg/day for 7 days in rats. Plasma, tissues, urine, and feces were collected from 0 to 24 h after dosing on days 1 and 7. The concentrations of asiaticoside, madecassoside, asiatic acid, and madecassic acid were simultaneously analyzed by liquid chromatography-tandem mass spectrometry. No significant change was observed in physical and blood biochemical parameters of the animals treated with ECa 233 for 7 days. The maximum plasma concentration and area under the curve at day 7 of madecassoside and asiaticoside decreased by 70–80% from day 1. However, both triterpenoid glycosides were extensively distributed and accumulated, resulting in significantly higher concentrations at pharmacologically relevant organs. Madecasssic acid and asiatic acid are major metabolites mainly found in and excreted via feces. Moreover, multiple dosing of ECa 233 increased mRNA expression of Abcb1a and Abcc2 in the small intestine by approximately 2- to 3-fold. This is the first study to identify an inductive effect of a standardized extract of C. asiatica after multiple oral dosing in rats. Potential drug-herb interactions when ECa 233 is coadministered with Abcb1a and Abcc2 substrates calls for further investigations.

In vitro effects of asiaticoside on the proliferation of and connective tissue growth factor expression by keloid-derived fibroblasts

[Objective] To investigate the m vitro effect of asiaticoside on the proliferation of and connective tissue growth factor (CTGF) expression by keloid-derived fibroblasts.[Methods] Tissue samples from patients with keloid were obtained for primary culture of fibroblasts.After 3 to 7 passages,the fibroblasts were incubated with different concentrations (100,10,1,0.1,0.01 mg/L) of asiaticoside or dimethyl sulfoxide (DMSO) for 24,48 and 72 hours followed by the determination of cell viability by methyl thiazolyl tetrazolium (MTT) assay.Immunohistochemistry and Western blot were carried out to quantify the expression of CTGF in the fibroblasts at 48 hours after treatment with different concentrations of asiaticoside.The morphology of fibroblasts was observed before and after the treatment with asiatieoside.[Results] As morphological observation showed,different concentrations of asiaticoside induced an obvious apoptosis and growth inhibition in fibroblasts.The growth of fibroblasts was suppressed by asiaticoside of 1-100 mg/L in a dose-dependent manner (r =0.95,0.90,0.92 for 24-,48- and 72-hour treatment respectively,all P ＜ 0.01 ),and one-factor analysis of variance revealed statistical differences in the growth inhibition rate in fibroblasts between different treatment durations for each tested concentration of asiaticoside (all P ＜ 0.01 ).There was a strong expression of CTGF in untreated fibroblasts,which was weakened by the treatment with asiaticoside for 48 hours.The number of CTGF-positive fibroblasts per 100 cells was 73 in untreated fibroblasts,significantly higher than that in fibroblasts treated with asiaticoside at 1 mg/L (54,t =4.34,P ＜ 0.01 ) and 10 mg/L (46,t =6.26,P ＜ 0.01 ),and statistical differences were observed between the fibroblasts treated with asiaticoside at 1 mg/L and 10 mg/L (t =1.95,P ＜ 0.05).Western blot also showed that the expression of CTGF was inhibited by 48-hour treatment with asiaticoside,and the inhibitive effect displayed a trend to increase with the rise in the concentration of asiaticoside.[Conclusion] Asiaticoside can effectively inhibit the oroliferation of and CTGF expression by fibroblasts in vitro. Key words: Keloid; Fibroblasts; Asiaticoside; Connective tissue growth factor

Identification of Major Active Ingredients Responsible for Burn Wound Healing ofCentella asiaticaHerbs

Centella asiatica herbs have been prescribed as a traditional medicine for wound healing in China and Southeast Asia for a long time. They contain many kinds of triterpenoid compounds, mainly including glycosides (asiaticoside and madecassoside) and corresponding aglycones (asiatic acid and madecassic acid). To identify which is the major active constituent, a comprehensive and comparative study of these compounds was performed. In vitro, primary human skin fibroblasts, originating from healthy human foreskin samples, were treated with various concentrations of asiaticoside, madecassoside, asiatic acid, and madecassic acid, respectively. Cell proliferation, collagen synthesis, MMP-1/TIMP-1 balance, and TGF-β/Smad signaling pathway were investigated. In vivo, mice were orally administered with the four compounds mentioned above for two weeks after burn injury. The speed and quality of wound healing, as well as TGF-β 1 levels in skin tissues, were examined. Interestingly, in contrast to prevalent postulations, asiaticoside and madecassoside themselves, rather than their corresponding metabolites asiatic acid and madecassic acid, are recognized as the main active constituents of C. asiatica herbs responsible for burn wound healing. Furthermore, madecassoside is more effective than asiaticoside (P = 0.0446 for procollagen type III synthesis in vitro, P = 0.0057 for wound healing speed, and P = 0.0491 for wound healing pattern in vivo, correspondingly).

Effect of Asiaticoside on 99mTc-Tetrofosmin and 99mTc-Sestamibi Uptake in MCF-7 Cells

This study was done to examine the effect of asiaticoside on MCF-7 cell uptake of (99m)Tc-tetrofosmin ((99m)Tc-Tfos) and (99m)Tc-sestamibi ((99m)Tc-MIBI). The 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the effect of a 50% inhibitory concentration of asiaticoside on MCF-7 cell proliferation. MCF-7 cells were treated with 10, 20, 30, 40, and 50 μM asiaticoside for 48 h and then incubated with 59.2 MBq of either (99m)Tc-Tfos or (99m)Tc-MIBI tracer for 60 min. The uptake of the tracers was measured with a dose calibrator. The 50% inhibitory concentration of asiaticoside for MCF-7 cells was determined with the MTT assay to be 40 μM. The uptake results were expressed as the mean ± SE radioactivity in MBq/mg of protein, and P values were also calculated (P values of 0.03 indicated significant differences). In the control (no asiaticoside) and at 10, 20, 30, 40, and 50 μM asiaticoside, the mean levels of (99m)Tc-Tfos uptake were 0.79 (SE, 0.059) (P = 0.14), 0.84 (SE, 0.057) (P = 0.60), 0.47 (SE, 0.034) (P = 0.03), 0.40 (SE, 0.050) (P = 0.03), 0.37 (SE, 0.050) (P = 0.03), and 0.15 (SE, 0.023) (P = 0.03), respectively; the mean levels of (99m)Tc-MIBI uptake were 0.95 (SE, 0.007) (P = 0.14), 0.81 (SE, 0.009) (P = 0.60), 0.79 (SE, 0.019) (P = 0.03), 0.63 (SE, 0.004) (P = 0.03), 0.13 (SE, 0.006) (P = 0.03), and 0.07 (SE, 0.008) (P = 0.03), respectively. Asiaticoside concentrations of 10, 20, 30, 40, and 50 μM revealed the uptake kinetics for both (99m)Tc-Tfos and (99m)Tc-MIBI in MCF-7 cells. (99m)Tc-Tfos and (99m)Tc-MIBI showed similar trends; the radioactivity uptake was dose dependent, and asiaticoside inhibited 16% and 47% of (99m)Tc-Tfos uptake and (99m)Tc-MIBI uptake in MCF-7 cells, respectively. This study showed that asiaticoside, acting as a biochemical modulator, may induce apoptosis and enhance antitumor activity in MCF-7 cells, as determined by (99m)Tc-Tfos and (99m)Tc-MIBI uptake. These findings are promising for cancer chemotherapy. Future studies should be performed to confirm our findings and to further delineate the clinical role of asiaticoside.

Measurement and Correlation of Solubilities of Asiaticoside in Water, Methanol, Ethanol,n-Propanol,n-Butanol, and a Methanol + Water Mixture from (278.15 to 343.15) K

The solubilities of asiaticoside in water, methanol, ethanol, n-propanol, n-butanol, and a methanol + water mixture were determined over the temperature range from (278.15 to 343.15) K by a static analytical method. The concentrations of asiaticoside in the saturated solutions were analyzed by reverse-phase high-performance liquid chromatography. An “N”-type curve was found for the solubilities of asiaticoside in the methanol + water mixture. Two empirical equations were proposed to correlate the experimental data, which fit the data well.

The cellular and molecular mechanism of inhibitory effect of asiaticoside on capsular contracture following breast augmentation

To explore the cellular and molecular mechanism of the inhibitory effect of asiaticoside on capsular contracture following breast augmentation. Contracture capsule derived fibroblasts were cultured in medium with different concentration of asiaticoside. The cell proliferation, collage synthesis and alpha-SMA expression were detected by means of 3H-thymidine incorporation, 3H-proline incorporation, and Western-blot. The results were analyzed by SPSS 11.0 with t test. DNA and collagen synthesis of fibroblasts were dramatically inhibited when the asiaticoside reached the concentration of 50 mg/L. The inhibitory rate was 34.7% and 30.1% respectively, showing a significant difference from that in control group (P < 0.05). The inhibitory effect increased with the rise of the asiaticoside concentration in a dose-dependent manner. When the concentration of asiaticoside reached 25 mg/L, the expression of alpha-SMA was down-regulated with an activation index of 1.673, showing a significant difference when compared with that in control group (P < 0.05). asiaticoside can effectively inhibit the DNA and collagen synthesis of capsule-derived fibroblasts. The trans-differentiation of fibroblast to myo-fibroblasts is also prevented by it.

Effects of asiaticoside on the expression of Smad protein by normal skin fibroblasts and hypertrophic scar fibroblasts

The precise mechanism of asiaticoside in molecular and gene expression levels of Smad protein and mRNA still remains unknown. We hypothesised that asiaticoside might inhibit the formation of hypertrophic scarring by affecting the expression of Smad protein and interfering with the Smad signalling pathway. To investigate the effects of asiaticoside on the expression of Smad protein by hypertrophic scar fibroblasts (HSFs) and to clarify the mechanism of asiaticoside in scar treatment. Normal skin fibroblasts and HSFs were exposed to various concentrations of asiaticoside in serum-free medium for 72 h, then immunocytochemistry was used to examine the localization and expression of phosphorylated Smad2 and Smad7. The expression of Smad protein was studied both at the transcriptional and post-transcriptional levels, using conventional reverse transcription PCR and Western blotting. Asiaticoside markedly enhanced the expression of inhibitory Smad7, but it had no effect on the expression of Smad2. Further study revealed that asiaticoside could induce Smad7 to enter the cytoplasm from the nucleus. Asiaticoside inhibits scarring probably by enhancing the expression of inhibitory Smad7, and is a potential treatment for scarring.

Standardization of pegagan extract (Centella asiatica (L.) Urban) based on siaticoside content using TLCdensitometricmethod

In assuring people consuming herbal products of high standard and reliable quality, a standardization process was performed on an Indonesian herb namely pegagan (Centella asiatica (L.) Urban) from Kaliurang, Boyolali, and Tawangmangu regions emphasizing on asiaticoside contents as its active constituent. Process of standardization was initiated by searching the best solvent for extraction. Water, ethanol 30%, 50%, and 70% were used for maceration of the herb. Asiaticoside contents were then determined by TLC-densitometry. The result of research showed that ethanol 70 % could dissolve highest concentration of asiaticoside from pegagan herb so it was then used for larger scale extraction of three different plant materials. TLC-densitometric data showed that asiaticoside contents of pegagan herb originally from Kaliurang were 0.98 ± 0.13.%, Boyolali were 1.34 ± 0.20.%, and Tawangmangu were 0.21 ± 0.3 %. Key words: Centella asiatica (L.) Urban, asiaticoside, TLC-densitometry

Inhibitory Effects ofCentella asiaticaWater Extract and Asiaticoside on Inducible Nitric Oxide Synthase During Gastric Ulcer Healing in Rats

In this study, the effects of Centella asiatica water extract (CE) and its active constituent, asiaticoside (AC), on the expression and activity of inducible nitric oxide synthase (iNOS) during gastric ulcer healing in rats were investigated. CE was prepared from Centella asiatica dry plant and the concentration of AC in CE was quantitatively determined with the use of high performance liquid chromatography analysis. Different concentrations of CE (0.10 g/kg and 0.25 g/kg) and AC (5 mg/kg and 10 mg/kg) were orally administered to rats with acetic acid-induced gastric ulcers. They were found to reduce the size of the ulcers at days 1, 3 and 7 after ulcer induction in a dose-dependent manner, with a concomitant attenuation of iNOS activity and protein expression at the ulcer tissues. The levels of nitrite and nitrate (NO(X)-), the stable end-products of nitric oxide (NO), in the gastric ulcer tissues were also decreased. N-[3-(aminomethyl)benzyl]acetamidine (1400W), a highly selective inhibitor of iNOS, was found to produce similar but more potent inhibition on iNOS activity at a dose of 0.1 mg/kg. These findings indicate that CE and AC have an anti-inflammatory property that is brought about by inhibition of NO synthesis and thus facilitates ulcer healing.

The healing effects of Centella extract and asiaticoside on acetic acid induced gastric ulcers in rats

In this study, the healing effects of Centella asiatica water extract (CE) and asiaticoside (AC), an active constituent of CE, on acetic acid induced gastric ulcers (kissing ulcers) in rats were examined. CE was prepared from Centella asiatica dry plant and the concentration of AC in CE was quantitatively determined with the use of high performance liquid chromatography analysis. Different concentrations of CE and AC were orally administered to rats with kissing ulcers. They were found to reduce the size of the ulcers at day 3 and 7 in a dose-dependent manner, with a concomitant attenuation of myeloperoxidase activity at the ulcer tissues. Epithelial cell proliferation and angiogenesis were on the other hand promoted. The expression of basic fibroblast growth factor, an important angiogenic factor, was also upregulated in the ulcer tissues in rats treated with CE or AC. These results further suggest the potential use of Centella asiatica and its active ingredient as anti-gastric ulcers drugs.

The morphological and pharmacological effects of asiaticoside upon skin in vitro and in vivo

The toxicity of asiaticoside to skin in vitro was investigated by a microrespiration technique and by histology. Compared with other therapeutic agents the toxicity of asiaticoside was not excessive. The effect of asiaticoside on the biosynthesis of skin mucopolysaccharides and collagen was also investigated in vitro. Toxic effects to both processes were evaluated; these were comparable to the effect of asiaticoside on skin respiration. Histology showed that certain concentrations of asiaticoside affected the epidermis, possibly increasing keratinisation. There was no evidence that asiaticoside stimulated either respiration or biosynthesis of ground substance. Asiaticoside failed to influence the mortality and healing of experimental burns in mice.