Herb Centella Asiatica Research Articles

Asiatic acid ameliorates doxorubicin-induced cardiotoxicity by promoting FPN-mediated iron export and inhibiting ferroptosis.

Doxorubicin (DOX), a common chemotherapeutic agent in cancer therapy, is accompanied by pronounced cardiotoxicity. Ferroptosis has been implicated in the pathogenesis and therapeutics of DOX-induced cardiotoxicity (DIC). Asiatic acid (AA), a pentacyclic triterpene from the Chinese medicinal herb Centella asiatica, displays antioxidant, anti-inflammatory, and antiapoptotic activities. In this study, we investigated the beneficial effects of AA against DOX-induced ferroptosis and cardiotoxicity and the underlying mechanisms. A chronic DIC model was established by challenging mice with DOX (5 mg/kg, i.p.) once per week for 4 weeks. Concurrent with DOX insult, the mice were administered AA (25 mg·kg-1·d-1, i.g.). Cardiac function and mechanical properties of isolated cardiomyocytes were evaluated at the end of treatment. We showed that AA administration preserved cardiac function, significantly reduced cardiac injury, and improved cardiomyocyte contractile function in DIC mice. The beneficial effects of AA were causally linked to the inhibition of DOX-induced ferroptosis both in vivo and in vitro. We revealed that AA attenuated DOX-induced iron accumulation in HL-1 cells by increasing FPN-mediated iron export, in a Nrf2-dependent manner. AA upregulated Nrf2 expression and promoted Nrf2 nuclear translocation in DOX-treated HL-1 cells. Moreover, AA-offered benefits against DOX-induced cardiac dysfunction and ferroptosis were abolished by Nrf2 inhibitor ML385 (30 mg·kg-1·d-1, i.p.) administrated 30 min before AA in DIC mice. Our data favor that AA promotes FPN-mediated iron export to inhibit iron overload and ferroptosis in DIC, suggesting its therapeutic potential in the treatment of DIC.

Computational analysis of phytocompounds in Centella asiatica for its antifibrotic and drug-likeness properties - Herb to drug study

Oral submucous fibrosis (OSMF) is a potentially malignant disorder with no permanent cure that affects the quality of life due to trismus. Computational pharmacology has accelerated the discovery of drug candidates for the treatment of incurable diseases. The present study aimed to screen the compounds of the miracle herb Centella asiatica with drug-likeness properties based on the absorption, distribution, metabolism, and excretion (ADME) properties. The pharmacological actions of these screened compounds against OSMF were identified by network pharmacology, gene ontology, pathway enrichment analysis, molecular docking, and simulation. Fifteen drug-like ligands were identified after virtual screening viz; asiatic acid, kaempferol, quercetin, luteolin, apigenin, bayogenin, gallic acid, isothankunic acid, madecassic acid, madasiatic acid, arjunolic acid, terminolic acid, catechin, epicatechin, and nobiletin. 850 potential targets were predicted for the ligands, which were analyzed against 354 proteins associated with OSMF. Compound pathway analysis and disease pathway analysis identified 53 common proteins. The GO enrichment analysis identified 472 biological process terms, 76 molecular function terms, and 44 cellular component terms. Pathway enrichment analysis predicted 142 KEGG pathways, 35 Biocarta pathways, and 236 Reactome pathways for the target proteins. The analysis revealed that the herb targets crucial events of fibrosis such as inflammation, oxidative stress, apoptosis, collagen deposition, and epithelial-mesenchymal transition. The common 53 proteins were used for protein-protein interaction (PPI) network analysis, which revealed 4 key proteins interacting with the phytocompounds viz; transforming growth factor-β1 (TGF-β1), mothers against decapentaplegic-3 (SMAD-3), mitogen-activated protein kinase-1 (MAPK-1) and proto-oncogene tyrosine-protein kinase (SRC). Molecular docking revealed that all ligands had a good binding affinity to the target proteins. Bayogenin had the highest binding affinity towards MAPK-1 (−9.7 kcal/mol), followed by isothankunic acid towards SRC protein (−9.3 kcal/mol). Madasiatic acid had the highest binding affinity to SMAD-3 (−7.6 kcal/mol) and TGF-β1 (−7.1 kcal/mol). Molecular dynamics simulation demonstrated stable ligand protein interactions of bayogenin and MAPK complex, isothankunic acid and SRC complex. This in silico study is the first to identify potential phytochemicals present in Centella asiatica and their target molecules, which might be responsible for reversing OSMF.

Timing of systemic resistance induced by local exogenous ABA application within clonal network of stoloniferous herb Centella asiatica subjected to low water availability.

Resistance traits of plants can be activated both at the damaged site and undamaged parts. Systemic resistance induced by local exogenous abscisic acid (ABA) application alleviated negative effect of low water availability on growth performance of clonal plant. However, timing of systemic resistance was poorly understood. Timing of systemic resistance refers to its activation and decay time within clonal network. Clonal fragment of Centella asiatica with four successive ramets (including first-oldest, second-older, third-old and fourth-young ramets) subjected to low water availability (20% soil moisture content) was used to explore effects of local exogenous ABA application on the timing of resistance activation and decay. Systemic resistance activated by local exogenous ABA application after 4 days remained at least 28 days. Compared with control, biomass accumulation of whole clonal fragment, root biomass and ratio of belowground to aboveground biomass significantly increased by local exogenous ABA application after 28 days. It is suggested that rapid activation and delay of resistance response induced by local exogenous ABA application within clonal network may improve fitness of clonal plant subjected to abiotic stress.

Screening of different extraction methods for maximum production of total flavonoids, tannins, and antioxidants from Centella asiatica

The study was carried out to maximise the extraction efficiency of different extraction methods and evaluate the total flavonoid, tannin, and antioxidant activity of the wellknown medicinal herb Centella asiatica. A total of nine extraction methods were used viz. squeezing of fresh leaves, extraction of fresh leaves and oven-dried powder using boiled water, ethanol, methanol, and acetone as solvents, respectively. Different in vitro assays were used to maximize the extraction of total flavonoid content, and total tannin content. The antioxidant activities in terms of total phenolic content (TPC), DPPH(1,1-Diphenyl-2- picrylhydrazyl) radical scavenging assay, and ferric reducing antioxidant power (FRAP) of C. asiatica were also evaluated. The extract from oven-dried powder using 70% acetone exhibited the maximum flavonoid (3.0533±0.1069 mg QE/g) and tannin content (0.7800±0.0100 mg TAE/g). The powder extract using 70% acetone also exhibited the highest total phenolic content (13.883±0.050 mg GAE/g), DPPH (70.630±2.310%) and FRAP value (166.670±2.260 mg AAE/100g). The acetone extract of dried C. asiatica powder was found to be the best extraction procedure for tannin, total flavonoids, and antioxidant production. The findings of this study might be helpful in extracting natural antioxidants from C. asiatica and improve the existing literature.

Potential of asiaticoside encapsulated alginate chitosan nanoparticles in the management of antiproliferative activity in C6 glioma cells

BackgroundAsiaticoside is a pentacyclic triterpenoid saponin constituent of the medicinal herb Centella asiatica, known for its neuroprotective, anticancer and other biological effects. The present study aims to develop and characterise asiaticoside encapsulated alginate chitosan nanoparticles (ACNPs) and evaluate their antiproliferative potential on C6 glioma cells. MethodsACNPs were prepared by the ionic gelation polyelectrolyte complexation technique and characterised by dynamic light scattering, scanning electron microscopy and transmission electron microscopy. Cytotoxicity and antiproliferative effect of ACNPs were studied on C6 glioma cells using MTT and EdU-assays. Apoptosis/necrosis in C6 glioma cells was determined by annexin V and acridine orange/ethidium bromide (AO/EtBr) assays. Intracellular ROS generation in C6 glioma cells exposed with ACNP was determined by DCFDA assay using flow cytometry. Cell cycle analysis in C6 glioma cells treated with ACNP was performed by flow cytometry. ResultsSynthesised ACNPs showed spherical shape with 200 nm particle size, good thermal stability, and an acceptable polydispersity index. ASI from synthesized ACNPs demonstrated 90% encapsulation efficiency (EE) and 10% drug release within 24 h. Upon ACNP treatment mBA cells exhibited good cell viability and intact cell morphology. However, C6 cells displayed dose-dependent cytotoxicity when treated with ACNP. Annexin V and acridine orange/ethidium bromide (AO/EtBr) assays revealed late apoptosis and necrosis (p < 0.05) in C6 glioma cells treated with ACNP. Likewise, ACNPs significantly augmented reactive oxygen species generation (p < 0.05) in C6 glioma cells. ACNP treatment also resulted in cell cycle arrest at the G1 phase with chromatin condensation in C6 glioma cells. ConclusionsThese findings suggest that ACNPs act as an antiproliferative agent against C6 glioma cells via an increased intracellular ROS pathway that promotes apoptosis/necrosis. This study throws light on more in-depth mechanistic aspects of managing brain disorders using C. asiatica and its phytoconstituents.

Formulasi Sediaan Gel Dari Ekstrak Etanol Herba Pegagan (Centella asiatica (L) Urb)

Indonesia is a rich country in terms of flora and fauna diversity. Indonesia's wealth with various types of flora is a highlight for science, one of which is the gotu kola herb (Centella asiatica (L) Urb). The chemical compound asiaticoside is part of the triperoids contained in the gotu kola herb which has the potential as antibacterial, antifungal, antioxidant and anti-cellulite on the skin, suitable for topical preparations such as gels. This study aims to formulate a gel preparation from the ethanolic extract of the gotu kola herb (Centella asiatica (L) Urb) which meets the requirements of a good gel. The formulation of the gel preparation was carried out with concentrations of 20%, 25%, and 30%. Extracted by maceration method using 70% ethanol and in a vacuum rotary evaporator to obtain a thick extract. Evaluation of the gel preparation was carried out by organoleptic test, homogeneity test, pH test, dispersibility test, adhesion test and irritation test. The results of the evaluation carried out with the three concentrations met the requirements of the organoleptic test, homogeneity test, pH test, adhesion test and irritation test, but the ethanol extract gel of the gotu kola herb at concentrations of 20%, 25% and 30% did not meet the requirements for the good dispersion test

Anti-excitotoxicity and neuroprotective action of asiaticoside encapsulated polymeric nanoparticles in pilocarpine rodent seizure model

Asiaticoside (ASI), an ursane-type triterpenoid saponin, isolated from the memory enhancing herb Centella asiatica, is known for its neuroprotective activities. Here, the anti-excitotoxicity and neuroprotective effects of ASI encapsulated alginate chitosan nanoparticles (ACNPs) were evaluated in pilocarpine (PC) induced seizure in mice model. ACNPs were prepared by ionic gelation-polyelectrolyte complex method and their physicochemical characterization was carried out by TEM, SEM, DLS, XRD, and FTIR. Subsequently, their encapsulation efficiency (EE), in vitro drug release, cell viability, seizure score, DNA fragmentation, and mRNA expression of regulatory stress markers were evaluated. Membrane permeability of ACNPs in the brain, histopathology, and biological TEM and SEM analyses were also carried out. TEM of ACNPs showed spherical morphology with a particle size of 200–400 nm. DLS of ACNPs displayed an average size of 486.2 nm with polydispersity index (PDI) of 0.567 and zeta potential of –14.1 mV. ACNPs achieved high EE (&gt;90%) and controlled release (10%). Biological evaluation studies revealed ACNPs as non-toxic to mouse neural stem cells (mNSCs). They displayed enhanced brain permeability and attenuated seizure. Our results confirmed ACNPs as effective in crossing the brain membrane barrier and mitigating seizure severity induced by PC.

Pharmacological Effects of Centella asiatica on Skin Diseases: Evidence and Possible Mechanisms.

The medicinal herb Centella asiatica (L.) Urban known as gotu kola has been reported to exhibit a wide range of pharmacological activities. In particular, a significant body of scientific research exists on the therapeutic properties of preparations of C. asiatica or its triterpenes in the treatment of skin diseases. The present study is aimed to provide a comprehensive overview of the beneficial effects of C. asiatica on skin diseases. Peer-reviewed articles on the potent dermatological effects of C. asiatica were acquired from PubMed, Web of Science, Scopus, ScienceDirect, and SciFinder. This review provides an understanding of pharmacological studies which confirm the potent dermatological effects and underlying molecular mechanisms of C. asiatica. This medicinal plant and its triterpenes include asiaticoside, madecassoside, and their aglycones, asiatic acid and madecassic acid. These compounds exert therapeutic effects on dermatological diseases such as acne, burns, atopic dermatitis, and wounds via NF-κB, TGF-β/Smad, MAPK, Wnt/β-catenin, and STAT signaling in in vitro and in vivo studies. However, additional rigorously controlled long-term clinical trials will be necessary to confirm the full potential of C. asiatica as a therapeutic agent.

Asiatic acid alleviates ischemic myocardial injury in mice by modulating mitophagy- and glycophagy-based energy metabolism.

Myocardial infarction (MI) causes disturbances in myocardial energy metabolism, ultimately leading to a poor prognosis. Cytosolic glycogen autophagy (glycophagy) and mitochondrial autophagy (mitophagy) are upregulated in MI to optimize energy metabolism but to a limited extent. Asiatic acid (AA), a pentacyclic triterpene derived from the traditional Chinese herb Centella asiatica, displays anti-inflammatory, antioxidant, and antiapoptotic activities. AA has been found to alleviate focal cerebral and liver ischemic injury by reversing mitochondrial dysfunction. In this study, we investigated whether AA exerted cardioprotective effects against MI by activating glycophagy and mitophagy to improve the energy balance. In vitro cardioprotective effects were examined in neonatal mouse cardiomyocytes subjected to oxygen-glucose deprivation for 12 h. Treatment with AA (2-50 μM) significantly increased cell viability and improved the energy metabolism evidenced by increased ATP level and phosphocreatine/ATP ratio. In vivo cardioprotective effects were studied in a mouse model of MI. Administration of AA (5-125 mg·kg-1·d-1, ig) significantly reduced infarct size and ischemic myocardial injury, and improved cardiac function. AA treatment also promoted mitophagy and relieved mitochondrial edema evidenced by increased number of mitophagosomes in ischemic myocardium in vivo and increased mitochondria-light chain 3 (LC3)-II colocalization in ODG-treated cardiomyocytes in vitro. Mitophagy activation was accompanied by activation of the AMPK signaling pathway. Knockdown of AMPK abolished AA-activated mitophagy. Furthermore, we showed that glycophagy was upregulated in OGD cardiomyocytes evidenced by increased starch binding domain protein 1 (STBD1)-GABA type A receptor-associated protein-like 1(GABARAPL1) interaction and extracellular acidification rate, whereas AA treatment further promoted glycophagy accompanied by PI3K/Akt activation. PI3K inhibitor LY294002 or Akt inhibitor GSK690693 blocked the effects of AA on glycophagy and glycolysis. Finally, simultaneous inhibition of glycophagy and mitophagy abolished the cardioprotective effects and energy regulation of AA. These results demonstrate that AA protects ischemic cardiomyocytes by modulating glycophagy- and mitophagy-based energy metabolism through the PI3K/Akt and AMPK pathways.

Tailor-made LED Illumination inducing Metabolic Variations in Centella asiatica, an important medicinal herb

Plant growth and development are influenced by the growth as well as physiological conditions, age of the plant, environment and genetic factors. Photo-induced signalling plays a significant role in regulating the growth and metabolism of plants. Secondary metabolites can serve as a metabolic signature for light-induced responses. The current study is aimed at analyzing the quantitative yield of chlorophyll pigments, total phenolic content and asiaticoside in LED illuminated grown plants of the medicinal herb Centella asiatica. A light intensity of 163 μmol/m2/s and 9-hour photoperiod, combination light source induced a total yield of 991 μg/g dry wt. asiaticoside. The total phenolic content has been correlated with the RB ratio. The analysis of the experimental results further confirms that it is feasible to grow medicinal plants under LED illuminated conditions. Such plants can serve as elite crude drugs owing to the presence of elevated active metabolic content.

Prolonged Treatment with Centella asiatica Improves Memory, Reduces Amyloid-β Pathology, and Activates NRF2-Regulated Antioxidant Response Pathway in 5xFAD Mice.

The medicinal herb Centella asiatica has been long been used for its neuroprotective and cognitive enhancing effects. We have previously shown that two weeks of treatment with a water extract of Centella asiatica (CAW) improves cognition and activates the endogenous antioxidant response pathway without altering amyloid-β (Aβ) plaque burden. Here, we assess the effect of long-term treatment of CAW in the 5xFAD mouse model of Aβ accumulation. Four-month-old 5xFAD mice were treated with CAW in their drinking water (2 g/L) for three months at which point they underwent cognitive testing as well as analysis of Aβ plaque levels and antioxidant and synaptic gene expression. In order to confirm the involvement of the antioxidant regulatory transcription factor NRF2 on the effects of CAW on synaptic plasticity, neurons isolated from 5xFAD mice were also treated with CAW and the targeted inhibitor ML385. Three months of treatment with CAW improved spatial and contextual memory as well as executive function in 5xFAD mice. This improvement was accompanied by increased antioxidant gene expression and a decrease in Aβ plaque burden relative to untreated 5xFAD animals. In isolated neurons, treatment with ML385 blocked the effects of CAW on dendritic arborization and synaptic gene expression. These results suggest that prolonged CAW exposure could be beneficial in Alzheimer's disease and that these effects likely involve NRF2 activation. Moreover, these findings suggest that targeting NRF2 itself may be a relevant therapeutic strategy for improving synaptic plasticity and cognitive function in Alzheimer's disease.

Asiatic Acid Prevents Cognitive Deficits by Inhibiting Calpain Activation and Preserving Synaptic and Mitochondrial Function in Rats with Kainic Acid-Induced Seizure.

Cognitive impairment is not only associated with seizures but also reported as an adverse effect of antiepileptic drugs. Thus, new molecules that can ameliorate seizures and maintain satisfactory cognitive function should be developed. The antiepileptic potential of asiatic acid, a triterpene derived from the medicinal herb Centella asiatica, has already been demonstrated; however, its role in epilepsy-related cognitive deficits is yet to be determined. In this study, we evaluated the effects of asiatic acid on cognitive deficits in rats with kainic acid (KA)-induced seizure and explored the potential mechanisms underlying these effects. Our results revealed that asiatic acid administrated intraperitoneally 30 min prior to KA (15 mg/kg) injection ameliorated seizures and significantly improved KA-induced memory deficits, as demonstrated by the results of the Morris water maze test. In addition, asiatic acid ameliorated neuronal damage, inhibited calpain activation, and increased protein kinase B (AKT) activation in the hippocampus of KA-treated rats. Asiatic acid also increased the levels of synaptic proteins and the number of synaptic vesicles as well as attenuated mitochondrial morphology damage in the hippocampus of KA-treated rats. Furthermore, proteomic and Western blot analyses of hippocampal synaptosomes revealed that asiatic acid reversed KA-induced changes in mitochondria function-associated proteins, including lipoamide dehydrogenase, glutamate dehydrogenase 1 (GLUD1), ATP synthase (ATP5A), and mitochondrial deacetylase sirtuin-3 (SIRT3). Our data suggest that asiatic acid can prevent seizures and improve cognitive impairment in KA-treated rats by reducing hippocampal neuronal damage through the inhibition of calpain activation and the elevation of activated AKT, coupled with an increase in synaptic and mitochondrial function.

Madecassoside encapsulated in alginate chitosan nanoparticles exerts anti-excitotoxicity effects in pilocarpine-induced seizure

Madecassoside (MAD), a pentacyclic triterpenoid saponin, and its source herb Centella asiatica are best known for their memory enhancing and neuroprotective properties. The effects of MAD- and similar secondary metabolite-encapsulated nanoparticles on brain are least explored. Henceforth, we have developed MAD encapsulated alginate chitosan nanoparticles (MACNPs), and their therapeutic potential is studied against pilocarpine (PC) rodent seizure model. MACNPs were synthesized by ionic-gelation polyelectrolyte technique and characterized by DLS, SEM, TEM and XRD. MAD encapsulation and release profile of MACNPs were studied in vitro. MACNPs distribution in mice brain tissues was also evaluated by SEM and TEM. MACNPs offered a negative zeta potential, considerable hydrodynamic size and spherical morphology. They significantly reduced the number of animals experiencing the most severe seizures, showed enhanced bioavailability and membrane integrity. Protection of tissues by MACNPs on PC toxicity in mice was assessed by histopathology on challenged mice brain. mRNA overexpression of MAPK1, MAPK14 genes together with bcl2, caspase 3 and hspa1b for oxidative stress and inflammation was observed. However, regulated expression of mRNA in p53 gene was noted. Our results describing the anti-seizure profile, combined with its observed mRNA expression and biodistribution, strongly support MACNP as a therapeutic candidate for a diverse range of epilepsies and related stress-inflammation.

Heavy metal accumulation efficiency, growth and centelloside production in the medicinal herb Centella asiatica (L.) urban under different soil concentrations of cadmium and lead

The present study explores the effect of varying concentrations of soil cadmium (50−200 mg/kg soil) and lead (200−800 mg/kg soil) on glasshouse grown plantlets of Centella asiatica. The plantlets were fairly tolerant to soil cadmium and lower concentrations of lead in terms of biomass accumulation. Application of 200 mg/kg soil cadmium concentration, led to enhanced biomass/two-fold increase in shoot length and a significant accumulation of cadmium in shoots and roots. Specific activities of anti-oxidant enzyme systems such as superoxide dismutase, ascorbate peroxidase, and guaicol peroxidase were also found to be elevated under treated conditions. Interestingly, the ratio of cadmium concentration in roots: soil (bioaccumulation coefficient factor) was more than 1, Centella asiatica could be a potential bioaccumulator of cadmium. Also since translocation of cadmium from roots to shoots was less than 40 %, the leaves, accumulating more than twenty five fold more centellosides (in 200 mg/kg soil cadmium v/s control conditions) could also be potentially exploited for phytochemicals with minimum post-harvest processing. Real time PCR analysis also revealed higher population of centelloside biosynthetic gene transcripts such as squalene synthase, farnesyl pyrophosphate synthase, β amyrin synthase, as compared to control conditions. Plant growth, although tolerant to lower concentrations of soil lead could not effectively accumulate lead in roots, and centelloside contents was also found to be only marginally higher than control. This study reports the possibility of exploiting C. asiatica as potential plant systems that may be cultivated on cadmium contaminated soils to reap phyto-pharmaceutical benefits.

Year-to-year crop shifts promote weed diversity in tropical permanent rainfed cultivation

In the past decades, the expansion and modernisation of agriculture in the mountainous areas of Southeast Asia has had severe impacts on biodiversity, as the once species-rich forests were turned into homogeneous fields receiving ample external inputs. A common feature of permanent cropping with annual crops is the frequent change of crop choice, depending on market opportunities or other motives. However, the precise effect of crop shifts on weeds in tropical areas is largely unknown. In this study, we investigated the short-term effect of crop sequences on the diversity of weed communities in smallholder fields of Northern Thailand. Crop choices were upland rice, maize, fallow and young tree plantations with or without intercrop. We counted the number of crop shifts and the number of crops involved during a 3-years period preceding weed sampling. We showed that the number of crop shifts did not affect weed density and biomass. However, herbaceous species number and diversity (measured as Shannon index) increased by 36% and 46% respectively, while herbaceous species dominance decreased by 38%, in fields with yearly crop shifts compared to fields with no shifts in the previous three years. The effect of a particular crop on diversity, or the effect of intercropping with young trees, was weaker. It was likely due to the more variable resources (especially light) in fields with two crop shifts, allowing species with different niches to co-exist. Crop type and frequent crop shifts did not affect shrub and tree species number, diversity or dominance. Some species were strongly associated with fields with no crop shift in the sequence (e.g. the tree Antidesma velutinosum) or to fields with two crop shifts in the sequence (e.g. the herb Centella asiatica, the C4 grass Digitaria radicosa). Overall, this study showed that in this agronomical system, maintaining yearly crop shifts does not significantly affect weed abundance, but supports in-field plant species diversity, which is likely to impact the services provisioned by tropical mountainous agro-ecosystems.

Asiatic acid supplementation during the in vitro culture period improves early embryonic development of porcine embryos produced by parthenogenetic activation, somatic cell nuclear transfer and in vitro fertilization

Asiatic acid is a pentacyclic triterpene enriched in the medicinal herb Centella asiatica, and it has been suggested to possess free radical scavenging and anti-apoptotic properties. The purpose of the current study was to explore the effects of asiatic acid on porcine early-stage embryonic development and the potential mechanisms for any observed effects. The results showed that 10 μM asiatic acid supplementation during the in vitro culture period dramatically improved developmental competence in porcine embryos derived from parthenogenetic activation (PA), somatic cell nuclear transfer (SCNT) and in vitro fertilization (IVF). Further analysis revealed that asiatic acid attenuated H2O2-induced intracellular reactive oxygen species (ROS) generation. Notably, asiatic acid not only enhanced intracellular GSH levels but also attenuated mitochondrial dysfunction. Gene expression analysis revealed that asiatic acid upregulated expression of the antioxidant-related gene Sod-1 and the blastocyst formation related gene Cox-2, while downregulating expression of the apoptosis-related gene Caspase-9 in SCNT blastocysts. These results suggest that asiatic acid exerts beneficial effects on early embryonic development in porcine embryos and that asiatic acid may be useful for improving the in vitro production of porcine embryos.

A preliminary study: Centella asiatica extract modulates acetylcholine in the heart

Previous studies have demonstrated the cardioprotective effect of acetylcholine (Ach), and therefore is indeed a potential theraupetic target for cardiovascular disease. Recent studies have shown that Ach is not only important in the pathological but also physiological state of the heart. The herb Centella asiatica has long been used to cure a number of conditions in traditional Indian medicine. Previous studies have shown that the acetylcholinesterase inhibitory activity of the Centella asiatica extract is capable of increasing the Ach level in the brain. However, it is unclear whether it can affect Ach level in other organs, such as the heart. In this study, healthy adult male Wistar rats were allocated into groups treated with the positive control, Donepezil, or the aqueous extract at doses 200 mg/kg, 400 mg/kg, and 800 mg/kg. After 21 days, the whole heart was evacuated and weighed prior to further measurement for Ach content. The present study showed that long-term consumption of 200 mg/kg and 400 mg/kg of aqueous Centella asiatica extract could modulate the Ach content in the heart.

Inverted U-shaped response of a standardized extract of Centella asiatica (ECa 233) on memory enhancement

The herb Centella asiatica has long been considered a memory tonic. A recent review found no strong evidence for improvement of cognitive function, suggesting negative results were due to limitations in dose, standardization and product variation. We used a standardized extract of C. asiatica (ECa 233) to study behavioral, cellular and molecular effects on learning and memory enhancement. ECa 233 (10, 30, and 100 mg/kg) was given orally to normal rats twice a day for 30 days. We used the Morris water maze to test spatial learning and performed acute brain slice recording to measure changes of synaptic plasticity in the hippocampus, a core brain region for memory formation. Plasticity-related protein expressions (NR2A, NR2B, PSD-95, BDNF and TrkB) in hippocampus was also measured. Rats receiving 10 and 30 mg/kg doses showed significantly enhanced memory retention, and hippocampal long-term potentiation; however, only the 30 mg/kg dose showed increased plasticity-related proteins. There was an inverted U-shaped response of ECa 233 on memory enhancement; 30 mg/kg maximally enhanced memory retention with an increase of synaptic plasticity and plasticity-related proteins in hippocampus. Our data clearly support the beneficial effect on memory retention of a standardized extract of Centella asiatica within a specific therapeutic range.

Effects of clonal integration on photochemical activity and growth performance of stoloniferous herb Centella asiatica suffering from heterogeneous water availability

Clonal integration allows translocation of water between interconnected ramets, improving fitness of clonal plants growing in heterogeneous water availability. However, few studies have been conducted to investigate the effects of clonal integration on physiological responses of clonal plants suffering from heterogeneous water availability. A greenhouse experiment was conducted by using clonal fragments of the stoloniferous herb Centella asiatica with two successive ramets. The basal ramets were grown in patches with 40% soil moisture (high water availability), while the apical ramets were grown in patches with 40% (high water availability), 20% (medium water availability) and 10% soil moisture (low water availability) respectively. Stolons between the basal and apical ramets were severed (clonal integration impeded) or retained intact (clonal integration allowed). Our results showed that clonal integration relieved negative impact of water deficiency on the apical ramets in terms of photochemical activity and growth performance. Clonal integration modified biomass partitioning both for the apical ramets growing in low water availability and the basal ramets growing in high water availability. In addition, significant cost of clonal integration was observed in terms of growth performance of whole clonal fragments. A possible explanation is that increased biomass allocation to root may reduce capturing of light energy, producing negative effect on biomass accumulation of the basal ramets under high water availability contrast between patches. The results suggest that the net benefit of clonal integration may depend on the contrast of resource availability between interconnected ramets.

Protective effects of Asiatic acid against pelvic inflammatory disease in rats.

Asiatic acid (AA) is one of the major components of the Chinese herb Centella asiatica and exerts a variety of pharmacological activities. However, the pharmacological effects of AA on pelvic inflammatory disease (PID) remain unknown. The purpose of the present study was to investigate the therapeutic efficacy and potential mechanisms of AA on PID in rats. A total of 75 female Sprague Dawley rats were randomly divided into the following five groups: A control group; a PID group; a PID + AA 5 mg/kg group; a PID + AA 35 mg/kg group; and a PID + AA 75 mg/kg group. Changes in cytokine and chemokine levels, myeloperoxidase (MPO) activity, nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome and nuclear factor-κB (NF-κB) activation, oxidative stress and cleaved caspase-3 were measured. AA treatment significantly decreased the excessive production of cytokines and chemokines and suppressed MPO activity and the activation of NLRP3 inflammasome, NF-κB and caspase-3, as well as oxidative stress. These results suggest that AA exhibits potent anti-inflammatory and antioxidant effects in rats with pathogen-induced PID and that the mechanism of these anti-inflammatory effects may be associated with the suppression of NLRP3 inflammasome activation and the NF-κB pathway.