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Background: Animal models had always served as integral part in introducing newer drugs for epilepsy. India has long tradition of using herbal medicines. Centella asiatica is an ingredient of various ayurvedic preparations to treat brain related disorders like insanity, memory loss and epilepsy. This research is done to find out the usefulness of Centella asiatica in epilepsy. Methods: Ethanolic extract of the plant is prepared using Soxhlet apparatus. Experimental seizure is induced in albino mice by maximal electroshock method. Resulting seizure goes through different phases. Duration of the phase of hind limb extension was the measured data. Clinically used anti epileptic drugs can abolish hind limb extension. Effect is compared to that of standard drug phenytoin. Four different doses of the plant extract; 100mg, 200mg, 500mg and 1000mg per kilogram bodyweight was given orally. Statistical analysis of data was done by one way ANOVA and Dunnett test.Results: Ethanolic extract of Centella asiatica exhibited statistically significant protection from maximal electroshock seizures. All given doses of the extract had p<0.05 when compared to control.Conclusions: Centella asiatica is potential source of anti-epileptic drug. Detailed phytochemical studies and animal experiments are recommended.
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The studies have suggested that advanced glycation end products (AGEs) induce stress oxidative and inflammatorypathway, which results in chronic complication. Centella asiatica (CA) has been shown as a promising candidate forAGE inhibitor due to its ability of reducing AGE production. This study aims to explore the molecular docking ofCA active compound as an inhibitor of AGEs and receptor AGEs (RAGEs). The top three docking structures werepicked for molecular dynamic (MD) simulations. Based on MD simulation in this study, we found that CA activecompound had been proven to interact with AGEs and RAGE. AGEs bound to asiaticoside, madasiatic acid, andmadecassic acid with a binding energy of −11.8253, −10.6724, and −10.1462 kcal/mol, respectively. Nonetheless,Asn106, Asp324, Asp376, Tyr420, and Tyr500 of AGEs made a significant contribution to the complex of asiaticosideAGE, as well as those for the madasiatic acid AGE, which were Asn118 and Tyr500. RAGE bound to asiaticoside,asiatic acid, and isothankunik acid with a binding energy of −10.6125, −9.4469, and −9.1015 kcal/mol, respectively.CA active compounds, specifically asiatic acid, madasiatic acid, and madecassic acid, interacted with AGEs, whereasasiaticoside and isothankunik acid interacted with RAGE based on docking and model studies.
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@#Introduction: Diabetic nephropathy is one of the most common complications in diabetes Mellitus. Hyperglycemia and chronic inflammation cause abnormal oxidative stress deposition that leads to the decrease of glutathione peroxidase-1 (GPx1) and endothelial Nitric Oxide synthetase (eNOS). It was reported that Centella asiatica has an anti-hyperglycemic and anti-inflammatory effect. However little is known about Centella asiatica effect in the kidney of DM. The objective of this study was to know the effect of Centella asiatica extract on Kim-1 (marker of kidney damage), GPx1 and eNOS mRNA expression in the kidney of DM rat model. Methods: Wistar DM rat model was divided into 6 groups namely non-DM group, DM group , DM with captopril and another DM group treated with Centella asiatica with three different dosages (250, 500 and 1000 mg/kg BW). The treatment was given for 8 weeks. The Kim-1, GPx1 and eNOS expression was measured using semi-quantitative PCR. Results: The DM group showed higher Kim-1 kidney mRNA expression but lower GPx1 and eNOS kidney mRNA compare to those on the non-DM group. Administration of Centella asiatica improves the expression of Kim-1, GPx1 and eNOS kidney mRNA expression in DM rat model. Conclusion: Centella asiatica has the potential to prevent kidney damage in DM rat model by improving Kim-1, GPx1 and eNOS kidney mRNA expression.
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@#Introduction: Kidney ischemic-reperfusion injury (IRI) is the main cause of acute kidney injury (AKI) which leads to the inflammation epithelial apoptosis and interstitial fibrosis as the chronic consequenses. Centella asiatica (CeA) has been known to have various pharmacological effects such as, anti-inflammatory, antioxidant, anti-fibrosis, and, anti-apoptosis. We aimed to elucidate the role of CeA in inhibiting kidney injury and infammatory mediators due to kidney IRI. Methods: Kidney IRI were performed with bilateral renal pedicles clamping in Swiss background mice (3 months-old, 30-40 grams) for 30 minutes (IR group, n=6), then terminated at day 7 after operation. At the next day, the mice that have been underwent bilateral kidney IRI were administered per-orally with ethanolic extract of CeA (210 mg/kg of BW, CeA1 group, n=6, and 420 mg/kg of BW, CeA2 group, n=6). The Sham Operation (SO group, n=6) was used as control. At the day 7 after the surgery, the mice were sacrificed and the kidneys were harvested. The kidney was used to assess tubular injury, interstitial fibrosis, and macrophage number, and another kidney was used to assess the mRNA expression of TLR4. Data were quantified using SPSS 22. Results: Kidney IRI produced significantly higher tubular injury, interstitial fibrosis and macrophage number (p<0.05) compared to SO with upregulating TLR4 mRNA expression (p<0.05). CeA treatment attenuated the tubular injury, interstitial fibrosis, macrophage number, and TLR4 mRNA expression which obviously shown in higher-dose of CeA (p<0.05). Conclusion: CeA ameliorates tubular injury, kidney fibrosis, and inflammatory mediators due to kidney IRI.
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@#Introduction: Diabetes mellitus (DM) is a disease that can cause complications in the kidneys. Centella asiatica extracts have the potential to inhibit pancreatic, liver and kidney tissue damage. This study was intended to determine the potential of C. asiatica extract in inhibiting kidney damage in an animal model of DM. Methods: Male Wistar rats were used in 5 treatment groups namely non-DM, DM, and DM with C. asiatica extract Dose 1 (250mg / kg), Dose 2 (500mg / kg) and Dose 3 (1000mg / kg). Changes in body weight, blood sugar, serum urea, kidney weight, glycosuria, and urine volume were observed in all treatment groups. Results: There were no significant differences between treatment groups on changes in blood glucose concentration, body weight, and serum ureum. However, C. asiatica treated group showed significantly lower value of urine volume, glycosuria, and kidney weight compare to those on Non-DM and DM groups. Decrease in blood glucose, although not significantly different, affects glucose urine excretion. Conclusion: C. asiatica extract has the potential to inhibit kidney damage in rats with DM through prevent the increase of urine volume, glycosuria, and kidney weight.
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Objective: To evaluate the effect of standardized extract of Centella asiatica ECa 233 on inflammatory mediator production through cyclooxygenase-2 (COX-2), extracellular signal-regulated kinase 1/2 (ERK1/2) and nuclear factor-κB (NF-κB) pathway in keratinocyte HaCaT cells.Methods: HaCaT cells were treated with 0.1, 1, 10 and 100 μg/mL ECa 233 in the presence of lipopolysaccharide (LPS). Proinflammatory cytokines and prostaglandin E2 were assessed with ELISA. Western blotting was used to determine the inhibition of COX-2, ERK1/2 and NF-κB protein expression. Results: ECa 233 suppressed LPS-induced release of interleukin-1β, tumor necrosis factor-α, and prostaglandin E2. ECa 233 also inhibited COX-2, phosphorylation of ERK1/2 and the activation of NF-κB. Moreover, the formation of reactive oxygen species (ROS) was decreased in response to LPS-inflamed keratinocytes. Conclusions: ECa 233 inhibits LPS-stimulated production of inflammatory mediators in keratinocytes via suppressing ERK1/2 and NF-κB pathways. The suppressive effect of ECa 233 may be related to an inhibition of ROS production.
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Aims@#Acalypha indica (AI), Centella asiatica (CA), and Sesbania grandiflora (SG) are vegetables commonly used in traditional medicine in Asian countries to treat skin problems. In this study, we investigated their pharmacological activities relevant to wound healing and synergistic actions to provide an insight into a promising vegetable combination as a candidate treatment for wounds. @*Methodology and results@#The stimulatory, antioxidant, and antibacterial activities of aqueous (A) and methanol (M) extracts of all the three vegetables were assessed alone and in combination in normal human dermal fibroblast (NHDF) cells in vitro. CA-A (89.52%) and the combination of AI-A+CA-A (90.76%) produced the highest percentage of wound closure. AI-A exhibited the highest total phenolic content (TPC) (82.94 mg GAE/g) and moderate reducing activity (61.63 mM Fe (II)/mg) when assessed by ferric reducing antioxidant power (FRAP) assay. Free radical scavenging activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), and the combination of AI-A+CA-A exhibited scavenging activity at IC50 = 379.75 µg/mL and IC50 = 578.7 µg/mL, respectively. Pre-treatment of NHDF cells with CA-M at 100 µg/mL offered the highest protection against hydrogen peroxide. All single and combined vegetable extracts showed poor antibacterial properties against Gram negative and Gram positive bacterial species implicated in wound infection. Only AI-A+CA-A executed synergism in fibroblast migration when assessed via the combination index (CI). Furthermore, screening and identification of AI-A, CA-A, and CA-M via UHPLC (LC-MS/MS) system revealed that the major components responsible for all the tested bioactivities were phenolic groups such as simple polyphenols, flavonoids, polysaccharides, and triterpenes (asiaticoside and madecassosides). @*Conclusion, significance and impact of study@#The vegetable extracts of A. indica, C. asiatica, and S. grandiflora exhibited good bioactivities independently. However, only AI-A+CA-A showed synergism in combination to accelerate the migration of fibroblast and increase antioxidant activities. These findings demonstrate the potential formulation of combined vegetable extracts from the two species of A. indica and C. asiatica for optimum wound healing properties.
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Plants, MedicinalABSTRACT
Objective: To evaluate the effect of standardized extract of Centella asiatica ECa 233 on inflammatory mediator production through cyclooxygenase-2 (COX-2), extracellular signal-regulated kinase 1/2 (ERK1/2) and nuclear factor-κB (NF-κB) pathway in keratinocyte HaCaT cells. Methods: HaCaT cells were treated with 0.1, 1, 10 and 100 μg/mL ECa 233 in the presence of lipopolysaccharide (LPS). Proinflammatory cytokines and prostaglandin E
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Aims@#The oriental-based herbs Acalypha indica (AI), Centella asiatica (CA), and Sesbania grandiflora (SG) possess a broad range of undisclosed therapeutic activities which are edible and easily available throughout the year. To convert the herb extracts into a potential drug form, aqueous (A) and methanol (M) extracts of herbs were assessed alone and in combination for their antifungal-demelanising activity and nitric oxide (NO) immunomodulatory responses. A new bioactive synergistic and antagonistic assessments approach was made on these herbs to identify which extract combination qualifies as a natural drug candidate.@*Methodology and results@#Via micro-dilution technique, methanol extract of A. indica (AI-M) showed the strongest antifungal activity against Aspergillus niger, with a minimum inhibitory concentration (MIC) of 50 mg/mL and a minimum fungicidal concentration (MFC) of 100 mg/mL. Sublethal (50 mg/mL) and subinhibitory (25 mg/mL) doses of AI-M produced the optimal black pigmentation reduction to demelanise A. niger. The combinations AI-M+CA-M, AI-M+SG-M, and CA-M+SG-M showed similar antifungal activities (MIC = 100 mg/mL). At 500 µg/mL, CA-A and the combination CAA+SG-A successfully induced RAW264.7 cells to produce NO at 17.85 µM and 40.84 µM, respectively. The combination of herbs extract showed synergistic interaction towards stimulation of NO production. In contrast, they demonstrated antagonism towards antifungal-demelanising properties. Compound identification of AI-M, SG-M, and SG-A were performed using a UHPLC-QTrap-MS/MS system, which detected phenolic compounds from various groups (cinnamic acids, benzoic acids, and flavonoids).@*Conclusion, significance and impact of study@#The combination of herb extracts showed better stimulation of NO production while the single herb extracts demonstrated good antifungal-demelanising activity. These findings help in the selection of herbs combination for potential natural drug discovery. A good combination of herbs demonstrated synergism to execute better bioactivities compared to individual herb extracts.
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Centella asiatica is an important medicinal plant which contains various phytocompounds. Asiatic acid and asiaticosideare two major compounds which are responsible for its various pharmaceutical activities. The present study analyzesthe effect of elicitor, i.e., methyl jasmonate on the synthesis of asiaticoside and asiatic acid (ATA) in shoot, callus, andcell suspension cultures of C. asiatica. A high-performance liquid chromatography analysis showed that the elicitationwith 100 µM concentration of methyl jasmonate enhanced asiaticoside content by 69-fold in callus culture, 39-fold inshoot cultures, and ATA by 1.9-fold in cell suspension culture. Thus, elicitation with methyl jasmonate is an effectivemethod of increasing the rate of biosynthesis of asiaticoside and ATA in plant cell cultures of C. asiatica
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Objective: Centella asiatica (L.) Urb from Umbelliferae is a potential source of secondary metabolites having immense medicinal value. Asiaticoside is the major therapeutic compound. In the present study, Identification of a possible relationship between concentration/transcript level expression of asiaticoside and concentrations of growth hormones at different growth stages was observed. The current study includes molecular and biochemical evaluation of stress generated in C. asiatica at different time intervals in vitro. Methods: The enhancement in auxin, cytokinin and final asiaticoside content were determined using immunoassay kits for auxin, cytokinin and HPLC analysis respectively. Transcript level expression at different growth phases was carried out using real-time RT-PCR. For isolation of stress-related miRNAs, reverse transcription of total RNA using miScript II RT Kit PCR System was carried out as per instructions. The differential expression of five selected miRNAs was done by Real-Time RT-PCR. The analysis of stress in vitro was done by quantification of Hydrogen Peroxide (H2O2), total phenolics and total antioxidants by H2O2 assay kit, total antioxidant assay kit and Folin Ciocalteau reagent respectively. The final asiaticoside content was determined by HPLC. Results: Differential expression of key genes involved in asiaticoside pathway showed significantly higher transcript expression, which is in correlation with the final asiaticoside content. The enhanced expression of miRNAs and the analysis of H2O2, total antioxidant capacity and total phenolics are suggestive of generation of oxidative stress under controlled conditions. Conclusion: The present study shows a direct correlation between oxidative stress and transcript/phytochemical estimation of asiaticoside content under in vitro conditions.
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@#Background: Chronic kidney disease (CKD) leads to inflammation, fibrosis and destruction of the renal architecture. Centella asiatica (CeA) is an herbaceous plant with antiinflammatory effects. We aimed to elucidate the effect of CeA on inflammation, fibrosis, vascular remodelling and antifibrotic substances in a 5/6 subtotal nephrectomy (SN) model in mice. Methods: Mice were divided into three groups: sham operation (SO, n = 6), 5/6 SN for seven days (SN7, n = 7) and SN7 with oral CeA treatment (SN7-CeA, n = 7). At day 7, mice were euthanised, kidneys were harvested and stained with periodic-acid Schiff (for tubular injury and glomerulosclerosis) and sirius red (for fibrosis and vascular remodeling) staining. mRNA expression of prepro-endothelin-1, nephrin, E-cadherin, bone morphogenic protein-7 (BMP-7), toll-like receptor 4 (TLR4), tumour necrosis factor-α (TNFα) and hepatocyte growth factor (HGF) were quantified using reverse transcriptase-PCR. Results: SN group demonstrated significant higher interstitial fibrosis, vascular remodeling, tubular injury and glomerulosclerosis (P < 0.01) compared to SO group. Meanwhile, in SN7-CeA demonstrated attenuation of vascular remodeling as shown by significant higher lumen area with lower Wall/Lumen area ratio compared to SN7. RT-PCR analysis showed up-regulation of nephrin, BMP-7 and E-cadherin mRNA expression (P < 0.05) and down-regulation of ppET-1 in SN7-CeA group compared to SN7 group (P < 0.05). Conclusion: CeA may ameliorate renal injury in the SN model in mice.
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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.
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Centella , Complex Mixtures , Inflammation , Malaysia , Memory , Plants , Rheumatic Diseases , Syphilis, Cutaneous , Wound HealingABSTRACT
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.
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Animals , Mice , Blotting, Western , Centella , Dermatitis , Dermatitis, Atopic , Ear , Enzyme-Linked Immunosorbent Assay , Immunoglobulin E , Immunoglobulins , Inflammation , Interleukin-6 , Interleukins , Nitric Oxide Synthase Type II , Prevalence , Skin , Tumor Necrosis Factor-alphaABSTRACT
Objective: To explore the effects of different drying methods on the quality of medicinal materials, and screen out the optimal drying process of Centella asiatica. Methods: The whole fresh grass of C. asiatica were dried by hot air, drying in the sun, drying in the sun and hot air combined, drying in the shade, microwave and vacuum respectively. Meanwhile, the drying time and rate were determined. The characters, identification, inspection, and leachable content of C. asiatica were analyzed by the method of pharmacopoeia. The content of asiaticoside, madecassoside, asiatic acid, madecassic acid, kaempferol-3-O-tutinoside, kaempferol, and quercetin were detected by HPLC analysis; The weighted scoring method was used to sort the comprehensive evaluation of multiple indexes. Results: Different drying methods consume different time, among which drying in the sun, shade and drying at 50 ℃ for more than 100 h, and the average drying rate was 24.83%. The effects of different drying methods on the properties of medicinal materials are mainly reflected in color and odor, among which 50-70 ℃ hot air drying had a better color, which was light green, and the odor of hot air drying and microwave drying at 80 ℃ and 85 ℃ also changed significantly. Although there were some differences in moisture and ash content, both of them met the pharmacopoeia standards. The drying method also had certain effects on the leachable, the maximum was 45.70%, and the minimum content of dry extract was 29.67%. The highest content of the total active ingredient was determined by HPLC using the method of drying in the shade, which was 83.032 mg/g, and the lowest was is 75 ℃ hot air drying, which was 40.982 mg/g. The highest total content of madecassoside and asiaticoside was 80 ℃ hot air drying, and the lowest was 75 ℃ hot air drying. Weighted score in the top three of line was 70 ℃, dried at 50 ℃ after drying in the sun, hot air drying at 50 ℃, and 85 ℃ hot air drying ranked the bottom. Conclusion: In summary, the suitable drying method for the production area of C. asiatica was 70 ℃ hot air drying.
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Aims: To investigate the physiological, leaf gas exchange and quality of Centella asiatica (pegaga) under different nitrogen fertilization rates. Study Design: Centella asiatica were exposed to four different nitrogen fertilization rates (0, 50, 100, 150 kg/ha) using urea (46% N) as nitrogen sources. The experiment was conducted in a randomized complete block (RCBD) design with three replications. Each treatment consisted of eight plants making the total of plants used in this study is 96 plants. Place and Duration of Study: Department of Biology, Faculty of Science, Universiti Putra Malaysia From May 2016-June 2016. Methodology: The growth parameters measured include total leaves numbers, leaf area, total chlorophyll content and total plant biomass. The carbon assimilation parameters were measured using LICOR 6400 XT Portable Photosynthesis System i.e net photosynthesis (A), Transpiration rate (E) and water use efficiency (WUE). Total phenolic and flavonoids contents from the leaves extracts were measured using Folin-Ciocalteu reagents. Results: The growth parameters such as leaves number, chlorophyll content, leaf area and total biomass were significantly influenced by nitrogen fertilization (P≤ 0.05), However, there were no significant difference observed between 50, 100 and 150 kg N/ha suggesting that 50 kg N/ha was the efficient rates to apply to enhance the growth of this plant. Meanwhile, the net photosynthesis (A) and water use efficiency (WUE) were enhanced with the increasing rate of nitrogen from 0>150 kgN/ha. The production of total phenolics and flavonoids was found to be highest under 100 kg/ha. The harvest index of total phenolics also showed the applications of 100 kg/ha gave the highest harvest index compared to the other nitrogen treatments. Conclusion: This study indicated growth and carbon assimilation parameters were enhanced under higher nitrogen fertilization and production of secondary metabolites was decreased with high rates of nitrogen. The recommended nitrogen fertilization for C. asiatica was at 100 kg N /ha, where it obtained the highest harvest index.
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Aims: This study was conducted to investigate the effect of salinity by using a different concentration of sodium chloride (NaCl), on growth, chlorophyll fluorescence and secondary metabolites production of Centella asiatica. Study Design: Centella asiatica plants were exposed to four different concentration of sodium chloride (0, 50, 100 and 150 mM). This research was conducted using a randomized complete block design 4 x 3 with three replications for each treatment and each treatment consists of 12 plants regarding four times harvesting. Place and Duration of Study: Glasshouse of SLAM field, University Agriculture Park, Universiti Putra Malaysia from February to April 2015. Methodology: Salinity stress was induced by irrigating the plants using four salinity levels (0, 50, 100 and 150 mM) of salt concentrations for 12 weeks. The leaves number were counted manually and the total plant biomass was taken by calculating the dry weight of root, stem, and leaf per seedling. The total chlorophyll content in the leaves was measured using a SPAD chlorophyll meter. Chlorophyll fluorescence was measured using Hansatech Pocket PEA, The leaf gas exchange were determined using a LI-6400XT portable photosynthesis system. Total phenolics and flavonoid was determined using Folin-Ciocalteu reagent. Phytochemical screening was conducted to determine the presence of tannin, terpenoids, phenolics, flavonoids, saponin, and alkaloids of plant samples under salinity stress. Results: Increased in salinity levels from 0 > 150 mM, the number of leaves, total biomass and total chlorophyll content were gradually decreased. Centella asiatica exhibit a significant decrease in net photosynthesis (A), transpiration rate (E), maximum efficiency of photosystem II (fv/fm) and Performance index (PI) when the salinity level increased. However, it was noticed that salinity stress significantly enhanced the total phenolic and flavanoid content of C. asiatica. It was also observed, that under salinity there were more presence of phytochemicals (tannin, terpenoids, phenolics, flavonoids, saponin and alkaloids) compared to the control. Conclusion: This study revealed that the increase in salinity level have greatly reduced the growth of C. asiatica but high salinity level also can enhance the production of secondary metabolites (total phenolic and flavonoid content) in C. asiatica.
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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.
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Animals , Male , Rats , Centella , Chemistry , Diabetes Mellitus, Experimental , Diabetic Nephropathies , Drug Therapy , Metabolism , Drugs, Chinese Herbal , Pharmacology , Kidney , Rats, Sprague-Dawley , Receptor, Transforming Growth Factor-beta Type I , Metabolism , Receptor, Transforming Growth Factor-beta Type II , Metabolism , Signal Transduction , Smad2 Protein , Metabolism , Smad3 Protein , Metabolism , Smad7 Protein , Metabolism , Transforming Growth Factor beta1 , MetabolismABSTRACT
Objective To optimize the preparation technology for Centella asiatica total glucosides (CTG) liposome and to investigate its percutaneous permeability in vitro. Methods Liposomes were prepared by reverse-phase evaporation technique. Taking the entrapment efficiency of madecassoside and asiaticoside as indexes, the preparation of liposome was optimized by using single factor and Box-Behnken response surface method. The properties of liposomes including morphology, entrapment efficiency, mean diameter, Zeta potential, and accumulative release were studied. Results The optimal formulation was lecithin-cholesterol (4:1), lecithin-CTG (23.22:1), and organic- aqueous (7:1). The liposome was smooth and spherieal in appearance, mean diameter, and Zeta potential were 201.7 nm and -15.7 mV, respectively. The entrapment efficiency of madecassoside and asiaticoside were 75.85% and 84.94%. The in vitro 12 h accumulative release was 52.10% and 45.97%. The equations for the permeation rate of madecassoside and asiaticoside in liposomes were Q = 67.93 t1/2-50.34, R2 = 0.988, Q = 139.74 t1/2-241.2, R2 = 0.987, respectively. The retention amount in the skin was 76 μg/cm2 and 48.7 μg/cm2, which were 1.56 and 1.18 times higher than those of solution. Conclusion The optimized process is rational, feasible, and good stability. The CTG liposome prepared in this study have the smaller size, the higher encapsulation efficiency, higher retain in skin and obvious sustained-release effects.
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Objective: To examine the effect of combination of Andrographis paniculata herb fraction (AHF) and Centella asiatica herb fraction (CHF) on PPARγ and GLUT4 mRNA expressions in 3T3-L1 adipocyte, and its effect on insulin-stimulated glucose uptake. Methods: 3T3-L1 adipocyte cells were used to investigate gene expression of PPARγ and GLUT4 proteins by reverse transcription-polymerase chain reaction method. The adipocyte cells were differentiated by using insulin, dexamethasone and 3-isobutyl-1-methylxanthine from 3T3-L1 cells. Pioglitazone, AHF, CHF and the combination of both herbs were evaluated on glucose uptake activity, PPARγ and GLUT4 mRNA expressions in 3T3-L1 adipocyte. Results: The results showed that combination of AHF at 30 μg/mL and CHF at 10 μg/mL could enhance insulin-stimulated glucose uptake. The combination also increased PPARγ and GLUT4 mRNA expressions significantly in comparison to those of negative control (DMSO). These effects were equal in comparison to those of pioglitazone (0.02 μM) and its single extracts. Conclusions: The combination of AHF and CHF can increase glucose uptake and insulin sensitivity through up-regulation of PPARγ and GLUT4 mRNA expressions in 3T3-L1 adipocyte.