Asiatic acid improves insulin secretion of β cells in type 2 diabetes through TNF- α/Mfn2 pathway / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effects and molecular mechanisms of asiatic acid on β-cell function in type 2 diabetes mellitus (T2DM).@*METHODS@#The T2DM model was established by high fat diet and streptozotocin injection in ICR mice, and the effects of asiatic acid on glucose regulation were investigated in model mice. The islets were isolated from palmitic acid-treated diabetic mice. ELISA was used to detect the glucose-stimulated insulin secretion, tumor necrosis factor (TNF)-α and interleukin (IL)-6. ATP assay was applied to measure ATP production, and Western blotting was used to detect protein expression of mature β cell marker urocortin (Ucn) 3 and mitofusin (Mfn) 2. The regulatory effects of asiatic acid on glucose-stimulated insulin secretion (GSIS) and Ucn3 expression were also investigated after siRNA interference with Mfn2 or treatment with TNF-α.@*RESULTS@#Asiatic acid with the dose of 25 mg·kg－1·d－1 had the best glycemic control in T2DM mice and improved the homeostasis model assessment β index. Asiatic acid increased the expression of Mfn2 and Ucn3 protein and improved the GSIS function of diabetic β cells in vitro and in vivo (both P<0.05). Moreover, it improved the ATP production of islets of T2DM mice in vitro (P<0.05). Interfering Mfn2 with siRNA blocked the up-regulation of Ucn3 and GSIS induced by asiatic acid. Asiatic acid inhibited islet TNF-α content and increased Mfn2 and Ucn3 protein expression inhibited by TNF-α.@*CONCLUSIONS@#Asiatic acid improves β cell insulin secretion function in T2DM mice by maintaining the β cell maturity, which may be related to the TNF-α/Mfn2 pathway.

Advances in the study of structural modification and biological activities of asiatic acid / 药学学报

Asiatic acid (AA) is a ursane pentacyclic triterpenoids, which possesses a wide range of pharmacological activities, such as anti-tumor, hypoglycemic, anti-inflammatory, anti-bacterial. Due to poor solubility and low bioavailability, clinical application of asiatic acid is limited. To address these defects, the structural modifications of AA have been carried out, and large numbers of AA-based derivatives with novel structure and eximious biological activity have been developed. In this paper, the research progress of structural modifications, biological activity, structure-activity relationship and mechanism studies in recent twenty years are reviewed, which provides reference for development of AA-related drugs.

Protective effect of asiatic acid on blood-retinal barrier in diabetic rats / 中华实验眼科杂志

Objective:To investigate the protective effect of asiatic acid (AA) on blood-retinal barrier (BRB) in diabetic rats and its possible mechanism.Methods:Ninety-six healthy 8-week-old male SD rats were randomly divided into normal control group, diabetes group, low-dose AA group and high-dose AA group, with 24 rats in each group.Intraperitoneal injection of streptozocin (STZ) was used to establish diabetes model.One month after the establishment of the model, the low-dose AA group and the high-dose AA group were given intragastrical administration of 37.5 mg/kg AA and 75.0 mg/kg AA, respectively, once a day according to grouping.The normal control group and the diabetes group were administrated with the same amount of 0.5% sodium carboxymethyl cellulose.The body weight of the rats were weighted at week 0, 1, 2, 3, 4 after intragastrical administration.Blood was taken from the tail vein and the blood glucose level was measured.The retina was obtained one month following the administration.Pathological changes of the rats retina were detected by hematoxylin-eosin (HE) staining.Evan's blue quantitative method was used to detect the damage of blood-retinal barrier (BRB). Immunofluorescence staining was performed to detect the distribution of Occludin, Notch1, Jagged canonical Notch ligand 1 (JAG1) and Delta like canonical Notch ligand 4 (DLL4) in retina.The mRNA and protein expressive levels of Occludin, Notch1, JAG1 and DLL4 were detected by Real-time PCR and Western blot.The study protocol was approved by a Scientific Research and Clinical Trial Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2020-KY-228). The use and care of animals complied with the Guide for the Care and Use of Laboratory Animals of National Institutes of Health and the 3R rules.Results:At 4 weeks after intragastrical administration, the body weight of the high-dose AA group was significantly higher than that of the diabetes group, and the blood glucose values were significantly lower in the high-dose AA group and the low-dose AA group in comparison with the diabetes group (all at P<0.05). The cells were arranged orderly with clear layered structure in the normal control group.In the diabetes group, the retina was thicker than that of the normal control group, with a thicker outer nuclear layer, disordered cell arrangement and unclear layered structure.Compared with the diabetes group, the total retinal thickness and structure were obviously improved in the low-dose AA group and the high-dose AA group.Evan's blue leakage in retina was (3.07±1.30), (13.73±3.88), (9.57±2.69) and (6.55±1.61)ng/mg in the normal control group, the diabetes group, the low-dose AA group and the high-dose AA group, respectively.There was a significant difference in leakage of Evan's blue among the four groups ( F=18.50, P<0.01), among which the leakage of Evan's blue dye in the high-dose AA group was significantly lower than that of the diabetes group ( P<0.01). Compared with the diabetes group, there was significantly higher relative expression level of Occludin protein and significantly lower relative expression levels of Notch1, JAG1 and DLL4 proteins in the other three groups (all at P<0.05). The relative expression level of Occludin protein was significantly higher and the relative expression levels of Notch1, JAG1 and DLL4 proteins were significantly lower in the high-dose AA group than those in the low-dose AA group (all at P<0.05). Compared with the normal control group, the Occludin mRNA expression level was significantly decreased and the expression levels of Notch1, JAG1 and DLL4 mRNA were significantly increased in the diabetes group and low-dose AA group (all at P<0.01). The Occludin mRNA expression level was higher and the Notch1 mRNA expression level was lower in the high-dose AA group than those in the diabetes group and the low-dose AA group, and the expression levels of JAG1 and DLL4 mRNA were lower in the high-dose AA group in comparison with the diabetes group, and the differences were statistically significant (all at P<0.05). Conclusions:Asiatic acid might play a protective role on BRB in diabetic rats by inhibiting Notch1 signaling pathway.

Microbial transformation of Asiatic acid by Syncephalastrum racemosum CGMCC 3.2500 / 药学实践杂志

Objective Asiatic acid is the main medicinal component of aursane pentacyclic triterpene and possessed various biological activities. In order to obtain better active Asiatic acid analogues, microbial transformation was used for structural modification. Methods Asiatic acid was biotransformed by Syncephalum racemosum CGMCC 3.2500. The structure of the compound was identified by high resolution electrospray ionization mass spectroscopy (HR-ESI-MS) and nuclear magnetic resonance spectroscopy (i.e., 1H NMR、13C NMR、1H-13C HSQC、1H-13C HMBC、1H-1H NOESY). Results The structure of the compound was determined as 2-oxo-3α, 15α, 23-trihydroxyurs-12-en-28-oic acid which was a new compound. Conclusion Syncephalum racemosum CGMCC 3.2500 can modify the structure of Asiatic acid and obtain Asiatic acid analogues.

Elicitation effect on the production of asiaticoside and asiatic acid in shoot, callus, and cell suspension culture of Centella asiatica

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

Asiatic Acid Protects Dopaminergic Neurons from Neuroinflammation by Suppressing Mitochondrial ROS Production

This study sought to evaluate the effects of Asiatic acid in LPS-induced BV2 microglia cells and 1-methyl-4-phenyl-pyridine (MPP⁺)-induced SH-SY5Y cells, to investigate the potential anti-inflammatory mechanisms of Asiatic acid in Parkinson’s disease (PD). SH-SY5Y cells were induced using MPP⁺ to establish as an in vitro model of PD, so that the effects of Asiatic acid on dopaminergic neurons could be examined. The NLRP3 inflammasome was activated in BV2 microglia cells to explore potential mechanisms for the neuroprotective effects of Asiatic acid. We showed that Asiatic acid reduced intracellular production of mitochondrial reactive oxygen species and altered the mitochondrial membrane potential to regulate mitochondrial dysfunction, and suppressed the NLRP3 inflammasome in microglia cells. We additionally found that treatment with Asiatic acid directly improved SH-SY5Y cell viability and mitochondrial dysfunction induced by MPP⁺. These data demonstrate that Asiatic acid both inhibits the activation of the NLRP3 inflammasome by downregulating mitochondrial reactive oxygen species directly to protect dopaminergic neurons from, and improves mitochondrial dysfunction in SH-SY5Y cells, which were established as a model of Parkinson’s disease. Our finding reveals that Asiatic acid protects dopaminergic neurons from neuroinflammation by suppressing NLRP3 inflammasome activation in microglia cells as well as protecting dopaminergic neurons directly. This suggests a promising clinical use of Asiatic acid for PD therapy.

Asiatic Acid Induces Apoptosis and Autophagy and Reduces MiR-17 and MiR-21 Expression in Pancreatic Cancer Cell Lines

This study investigated the cytotoxic effects and mechanism of action of asiatic acid in pancreatic cancer cell lines. First, we confirmed the cell viability of MIA PaCa-2 and PANC-1 cells after asiatic acid administration for 48 and 72 h. The viability of MIA PaCa-2 and PANC-1 cells decreased in a dose-dependent manner following asiatic acid administration. To investigate the underlying mechanism, we performed a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, annexin V assay, and western blotting. Asiatic acid induced apoptosis and autophagy through activation of AMP-activated protein kinase (AMPK) and inhibition of mammalian target of rapamycin (mTOR) in MIA PaCa-2 cells. Finally, the expression of miR-17 and miR-21, known as oncogenes in pancreatic cancer, was decreased by asiatic acid. These results indicate that asiatic acid has potential as a new therapeutic agent against pancreatic cancer.

Screening of Centella asiatica drying method based on comprehensive evaluation of quality / 中草药

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.

Inhibitory effect of asiatic acid on human liver carcinoma cells via cell cycle blockage and apoptosis induction / 中国肿瘤生物治疗杂志

@# Objective: To investigate the inhibitory effect of asiatic acid (AA) on malignant biological behaviors of human liver cancer cells and to explore the mechanism. Methods: Human liver cancer cell line (Huh7) was used as research subject, and treated with different concentrations of AA (0, 5, 10, 25, 50, 100 μmol/L) in vitro. The effect of AA on cell proliferation was determined by CCK-8 and EdU assay; the apoptosis and cell cycle distribution were detected by flow cytometry, while the expressions of apoptosis-related proteins (AKT, P-ERK 1/2 , p38, cleaved-caspase3, cleaved-caspase9, BAX, Bcl-2, AKT, ERK, p38, pro-caspase 3 and pro-caspase 9) were examined by WB. Results: AA could inhibit the proliferation of Huh7 cells in a dose- and time-dependent manner (all P<0.05). After being incubated with 10 μmol/L AA for 24 h, the proliferation of Huh7 cells was significantly inhibited (P<0.05), the apoptosis rate was significantly increased (P<0.05), and cell cycle was arrested in G1 phase (P<0.05).AAinduced p-p38 expression, but inhibited the expression of p-AKT and p-ERK in a dose-dependent manner (all P<0.05). In addition, as the concentration of AA increased, the levels of cleaved-caspase 3, cleaved-caspase 9 and BAX increased, while the level of Bcl-2 decreased (all P<0.05). Conclusion:AAinhibits the proliferation of human liver cancer cells and promotes its apoptosis, which is associated with the MAPK and PI3K/AKT pathways.

Inhibitory effect of asiatic acid on proliferation and apoptosis of TCA-8113 human tongue carel inoma cells / 中国药理学通报

Aim To investigate the effect of asiatic acid(AA) on the proliferation, apoptosis and cell cycle arrest of human tongue carcinoma TCA-8113 cells, and the possible mechanism. Methods The inhibitory effect of AA(0, 20, 30, 40, 50 pjnol • L"1) at different concentrations on TCA-8113 cells for 24 h was determined by MTT. The effect of AA on the colony formation rate of TCA-8113 cells was detected by colony formation test; The apoptosis of TCA-8113 cells by AA was detected by Hoechst 33342 staining and Annexin V-FITC/PI staining. Cell cycle changes were analyzed by flow cytometry. The expressions of protein in human tongue cancer cells, such as bcl-2, Bax, cleaved caspase-3, p53 and p21 proteins were detected by Western blot. Results The inhibitory rate of AA on TCA-8113 cells was concentration-dependent (P < 0.05), with the IC50 concentration of 42. 13 (xmol • L- 1, and the ability of cell colony formation decreased. The apoptosis rate increased with the increase of AA concentration (P < 0. 05). The concentration of AA 30 u.mol -L"1 gradually blocked the cell cycle in G2/M phase. Western blot analysis showed that the expressions of p53, p21 and Bax increased, while Bcl-2 decreased in a dose-dependent effect (P<0. 05). Conclusions AA can significantly inhibit the proliferation and apoptosis of human tongue carcinoma TCA-8113 cells, and its mechanism may be related to the regulation of p53 pathway related proteins p53, p21, Bax and the inhibition of Bcl-2 expression, and AA can arrest TCA-8113 cells cycle in G2/M phase.

Asiatic acid inhibits LPS-induced inflammatory response via Cardiology, regulating TLR4 and PPAR-γin VSMCs / 中国药理学通报

Aim To observe whether asiatic acid ( AA) can inhibit lipopolysaccharide ( LPS )-induced inflammatory response in VSMCs , and explore its mechanism of action .Methods The VSMCs isolated from aorta of SD rats were primarily cultured . The effect of AA on the cell viability of VSMCs was meas-ured by MTT assay .The protein and mRNA expression of IL-6, MCP-1, and TNF-α, were measured by ELISA assay and real-time PCR, respectively.The protein and mRNA of TLR4 and PPAR-γwere meas-ured by Western blot and real-time PCR, respectively . Results AA exhibited no effect on cellular viability between the concentration from 0 to 30 μmol · L-1 . After treating VSMCs with LPS (500μg· L-1 ) for 6h or 24 h, the protein and mRNA expression of IL-6, MCP-1, TNF-α, and TLR4 significantly increased ( P<0.05 );and on the contrary , the activity of PPAR-γwas significantly reduced ( P<0.05 ) .Treatment with AA (10, 20, 30 μmol· L-1 ) could concentration-de-pendently inhibit LPS-induced protein and mRNA ex-pression of IL-6, MCP-1, TNF-α.AA could also re-duce LPS-induced protein and mRNA expression of TLR4, and pretreatment of the cells with TLR4-siRNA could reduce LPS-induced inflammation . Moreover , treatment with AA could up-regulate the mRNA and protein expression of PPAR-γin VSMCs; however , GW9662 , a PPAR-γantagonist , partially attenuated AA' s anti-inflammatory effect .Conclusion AA can significantly inhibit LPS-induced mRNA and protein expression of IL-6, MCP-1, TNF-α, in VSMCs, which is partially dependent on suppressing TLR 4 and up-regulating PPAR-γ.

Effects of asiatic acid on MPTP-induced Parkinson's disease-like motor symptoms in mice / 中成药

AIM To investigate the effects of asiatic acid (AA) on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD)-like motor symptoms in mice and its neuroprotective mechanism.METHODS Forty-five male C57BL/6 mice,except the nine mice in control group,were induced to be the PD models by peritoneal injection of 25 mg/kg MPTP for seven days and then were randomly assigned to model group,low-dose,high-dose AA groups and positive control group.Both the control group and the model group were administered with 0.5％ sodium carboxyl methyl cellulose (CMC-Na) solution,the AA groups were dosed with 12.5 mg/kg and 25 mg/kg AA,respectively,and the positive control group was given 75 mg/kg daily intragastric gavage of levodopa for eleven days.On the twelfth day,behavioral tests were performed.Tyrosine hydroxylase (TH) positive cells in substantia nigra were detected by immunohistochemistry.The mRNA expressions of iNOS,COX-2,TNF-α,IL-1β,and malonaldehyde (MDA) content in midbrain were measured.The levels of IL-1 β and TNF-α in the serum were detected using ELISA kits.RESULTS The mice treated with asiatic acid performed better in behavior tests than those in the model group (P ＜0.05,P ＜0.01).In addition,asiatic acid effectively protected the dopaminergic neurons in the substantia nigra due to upregulated TH expression and increased number of TH positive cells (P ＜ 0.05).The asiatic acid-treated mice had their mRNA expressions of IL-1β,TNF-α,iNOS and COX-2 in midbrain markedly suppressed (P ＜0.05,P ＜0.01),and a significant MDA level decrease in the midbrain tissue as well (P ＜ 0.01).Furthermore,reductions of IL-1 β and TNF-α contents in the serum were observed (P ＜ 0.05,P ＜ 0.01).CONCLUSION Asiatic acid attenuates motor dysfunction and dopaminergic neuronal deficits in PD mice,and the neuroprotective mechanisms may attribute to its anti-oxidative and anti-inflammatory activities.

Asiatic acid enhances the chemosensitivity of U87MG glioma cells to paclitaxel through inhibiting the expression of drug resistance related proteins / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the inhibitive effect of asiatic acid (AA) on paclitaxel (PTX)-resistant glioma cells and its possible mechanism. Methods: The effects of AA on the proliferation and apoptosis of glioblastoma U87MG cells were detected by CCK-8 assay, Real-time quantitative polymerase chain reaction (qPCR) and Western blotting. The drug-resistant glioma cell line PR-U87MG was established by culturing the cells in concentration-increasing PTX. With U87MG cells as control, the PTX-resistance of PR-U87MG cells was confirmed using CCK-8 assay, and the mRNA and protein levels of MDR1 and LRP were measured with qPCR and western blotting. PR-U87MG cells were treated with AA, PTX or AA+PTX, and then the cell viability and apoptosis of each group were measured with CCK-8 assay, qPCR and Western blotting. Results: PTX-resistant PR-U87MG cell line was successfully established. AA inhibited the viability of U87MG and PR-U87MG cells in a dose-dependent manner (P<0.01) and significantly promoted their apoptosis (P<0.01). Compared with the group treated with AA or PTX alone, the group treated with the combination of AA and PTX had significantly decreased protein levels of PARP1 (P<0.01), drug-resistant related proteins (Pgp-1 and LRP [lung resistance protein], all P< 0.01), and markedly increased caspase 3 (P<0.01). Conclusion: AA could effectively enhance the sensitivity of U87MG cells to PTX, and the mechanism may be related to the suppressed expression of drug efflux-associated proteins Pgp-1 and LRP.

Determination of triterpenes in Melastoma dodecandrum Lour. from different producing areas by HPLC / 国际中医中药杂志

Objective To develop a HPLC method for the simultaneous content determination of three terpenoids inMelastoma dodecandrum Lour. (asiatic acid, betulinic acid, oleanolic acid).Methods The chromatographic column was set with waters SunFire C18 column (4.6 mm×150 mm, 5μm); the moving phase was acetonitrile -0.1%H3PO4; the column temperature was 30℃; the detection wavelength was 200 nm; the flow rate was 0.6 ml/min; and the sample volume was 25μl.Results A good linear relationship was observed in the range of 0.310-6.200μg (r=0.9999), 0.405-8.100μg (r=0.9999), 0.169-3.375μg (r=0.9998) for asiatic acid, betulinic acid, oleanolic acid respectively, with the average recovery rates of 102.08%, 101.81%, 102.22%. Conclusions The established method is convenient and sensitive, repeatability and stability, quality evaluation for Melastoma dodecandrum Lour.

Preparation and pharmacokinetic study of asiatic acid loaded chitosan-deoxycholic acid polymeric micelles in rats / 中草药

Objective To prepare asiatic acid (AA) loaded chitosan-deoxycholic acid self-assembled micelles (AA-CS-DCA PMs) adopting chitosan-deoxycholic acid (CS-DCA) as carriers and investigate its pharmacokinetic characteristics in rats. Methods AA-CS-DCA PMs were prepared by ultrasonic dispersion method. The characteristics of micelles were evaluated by the distribution of particle size, Zeta potential, drug loading, encapsulation efficiency, and in vitro release. Model of bile drainage was established in conscious rats and pre-column derivatization HPLC method was used to determine the concentration of AA in bile. Moreover, the pharmacokinetics characteristics of AA-CS-DCA PMs in vivo was evaluated by tmax, Cmax and AUC0-t. Results The particle size was (70.5 ± 9.8) nm, the Zeta potential was (38.4 ± 0.8) mV, and encapsulation efficiency and drug loading were (77.8 ± 1.2)% and (11.7 ± 0.2)%, respectively. The in vitro release profile showed a sustained release property. In vivo study showed that Cmax of AA-CS-DCA group (26.05 ± 3.04) μg/h was 2.8 times higher than that of the control group (9.19 ± 1.12) μg/h; The tmax of AA-CS-DCA PMs group prolonged significantly (P < 0.05) in biliary excretion (2 h vs 1 h) and the elimination half-life t1/2 was 1.8 times of the control group [(2.68 ± 1.71) h vs (1.49 ± 0.38 h)]. In addition, the AUC0-24 h which reflected the degree of drug absorption increased by 200% compared with the control group [(99.05 ± 12.83) μg vs (33.56 ± 8.33) μg]. Conclusion The chitosan- deoxycholic acid self-assembled micelles can raise the concentration of AA and prolong the retention time in vivo, which effectively improve the oral bioavailability of AA.

The recovery and protective effects of asiatic acid on differentiated human neuroblastoma SH-SY5Y cells cytotoxic-induced by cholesterol

Objective:To investigate the effect of asiatic acid (AA) on the differentiated human neuroblastoma SH-SY5Y cells cytotoxic-induced by cholesterol.Methods:Human neuroblastoma SH-SY5Y cells were either exposed to different concentrations of AA or treated with different doses of cholesterol to reveal their responding viability by MTT assay.The selective 1 μmol/L concentration of AA was then used to test for either the protective or the recovery effects on the cells treated with 250 μmol/L concentration of cholesterol.Results:AA has a propensity to directly increase the viability of differentiated human neuroblastoma SH-SY5Y cells.Cholesterol has significant cytotoxic effect on those cells in a concentration-dependent manner.AA has the ability to slightly recover the viability of the differentiated culture cytotoxic-induccd by cholesterol but could not protect those cells from cytotoxic-induced by cholesterol.Conclusions:High concentrations of cholesterol were observed to be harmful to the neurons and AA had a slight effect of reducing neuronal death caused by cholesterol.

The recovery and protective effects of asiatic acid on differentiated human neuroblastoma SH-SY5Y cells cytotoxic-induced by cholesterol

Objective To investigate the effect of asiatic acid (AA) on the differentiated human neuroblastoma SH-SY5Y cells cytotoxic-induced by cholesterol. Methods Human neuroblastoma SH-SY5Y cells were either exposed to different concentrations of AA or treated with different doses of cholesterol to reveal their responding viability by MTT assay. The selective 1 μmol/L concentration of AA was then used to test for either the protective or the recovery effects on the cells treated with 250 μmol/L concentration of cholesterol. Results AA has a propensity to directly increase the viability of differentiated human neuroblastoma SH-SY5Y cells. Cholesterol has significant cytotoxic effect on those cells in a concentration-dependent manner. AA has the ability to slightly recover the viability of the differentiated culture cytotoxic-induced by cholesterol but could not protect those cells from cytotoxic-induced by cholesterol. Conclusions High concentrations of cholesterol were observed to be harmful to the neurons and AA had a slight effect of reducing neuronal death caused by cholesterol.

Asiatic acid inhibits lung cancer cell growthandby destroying mitochondria

Asiatic acid (AA), a pentacyclic triterpene found in, displays significant anti-proliferative effects on cancer cellsalthough the underlying mechanism of this effect remains unknown. This study investigated the efficacy and mechanism of action of AA against lung cancer bothand. Using the MTT assay, AA was found to induce apoptosis in a dose- and time-dependent manner, an effect enhanced by pretreatment with an autophagy inhibitor. It also elevated expression of microtubule-associated protein 1 light chain 3 (LC3) and decreased the expression of p62. Furthermore, exposure to AA resulted in collapse of the mitochondrial membrane potential and generation of reactive oxygen species (ROS), suggesting mitochondria are the target of AA. In the mouse lung cancer xenograft model, oral administration of AA significantly inhibited tumor volume and weight accompanied by significant apoptosis of lung cancer cells. In addition, it led to a significant decrease in the expression of proliferating cell nuclear antigen (PCNA). In summary, the results show that AA significantly reduces lung cancer cell growth bothandand that the associated apoptosis is mediated through mitochondrial damage.

Oral absorption of asiatic acid nanoparticles modified with PEG / 中国中药杂志

A solvent diffusion method was used to prepare pegylated asiatic acid (AA) loaded nanostructured lipid carriers (p-AA-NLC), and the ligated intestinal circulation model was established to observe the absorption and distribution in small intestine. The concentration of AA in bile after oral administration of p-AA-NLC was detected by HPLC in healthy SD rats to indirectly evaluate the oral absorption promoting effect of PEG-modified namoparticles. The results showed that the penetration of p-AA-NLC was enhanced significantly and the transport capacity was increased greatly in small intestinal after PEG modification. As compared with the normal nanoparticles (AA-NLC), the Cmax of the drug excretion was increased by 76%, the time to reach the peak (tmax ) was decreased and the elimination half-life t1/2 was doubled in the rats after oral administration of p-AA-NLC, and the AUC0→t was 1.5 times of the AA-NLC group, indicating that the oral bioavailability of AA-NLC was significantly improved by hydrophilic modification of PEG.

Chemical constituents from leaves of Ilex chinensis / 中草药

Objective: To investigate the chemical constituents from the leaves of Ilex chinensis. Methods: The constituents were isolated and purified by various chromatographic methods, and the structures were elucidated by spectroscopic analysis. Results: Fifteen triterpenoid compounds were isolated from 70% ethanol extract in the leaves of I. chinensis and identified as 2α,3β-dihydroxy-23-norolean-4(24)-12(13)-dien-28-oic acid (1), 29-hydroxyhederagenin (2), ilexosapogenin A (3), rotundic acid isopropylidene (4), 2α,3β-dihydroxy-23-norurs-4(24)-12-dien-28-oic acid (5), 23-hydroxyursolic acid (6), 3β,23-dihydroxyursa-12,18- dien-28-oic acid (7), asiatic acid (8), 28-O-β-D-glucopyranoside siaresinolic acid (9), 28-O-β-D-glucopyranoside spathodic acid (10), 28-O-β-D-glucopyranoside pomolic acid (11), rotungenoside (12), 3β,23-dihydroxyurs-12,18-dien-28-O-β-D-glucopyranosyl ester (13), 3β,23-dihydroxyurs-12,19-dien-28-O-β-D-glucopyranosyl ester (14), and 3β,23-dihydroxyurs-12,19(29)-dien-28-O-β-D- glucopyranosyl ester (15). Conclusion: Compounds 1 and 2 are isolated from the plants of Ilex L. for the first time. All compounds are isolated from this plant for the first time.