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SUMMARY: We evaluated the role and mechanism of acteoside in the regulation of memory impairment induced by chronic unpredictable mild stress (CUMS). CUMS was used to induce depression in rats and the successful establishment of CUMS model were verified by forced swimming test and sucrose preference test. The Y-maze test and novel object recognition test assessed memory functions. The structural changes in the cortex and hippocampus were observed by hematoxylin and eosin (HE) staining. Immunofluorescence staining and western blotting determined the protein levels. Y-maze test and novel object recognition test showed that there was memory performance impairment in rats of CUMS group, which was improved by the acteoside treatment. HE staining showed that CUMS exposure damaged the structure in the cortex and hippocampus, while the acteoside treatment alleviated the structural changes. Compared with the control group, the levels of BNDF and CREB in the cortex and hippocampus of the CUMS group were significantly decreased. Acteoside significantly reversed the expressions of these proteins in CUMS rats. Meanwhile, compared with the control group, the levels of p-mTOR and p- P70S6K in the cortex and hippocampus of the CUMS group were significantly increased, and these changes were significantly reversed by acteoside. Nevertheless, the effect of acteoside on mTOR signaling was markedly blocked by rapamycin, a specific inhibitor of mTOR signaling. Acteoside can attenuate memory impairment and ameliorate neuronal damage and synaptic plasticity in depression rats probably via inhibiting the mTOR signaling pathway. Acteoside may serve as a novel reagent for the prevention of depression.
Evaluamos el papel y el mecanismo del acteoside en la regulación del deterioro de la memoria inducido por estrés leve crónico impredecible (ELCI). Se utilizó ELCI para inducir depresión en ratas y el establecimiento exitoso del modelo ELCI se verificó mediante una prueba de natación forzada y una prueba de preferencia de sacarosa. La prueba del laberinto en Y y la prueba de reconocimiento de objetos novedosos evaluaron las funciones de la memoria. Los cambios estructurales en la corteza y el hipocampo se observaron mediante tinción con hematoxilina y eosina (HE). La tinción por inmunofluorescencia y la transferencia Western determinaron los niveles de proteína. La prueba del laberinto en Y y la prueba de reconocimiento de objetos novedosos mostraron que había un deterioro del rendimiento de la memoria en ratas del grupo ELCI, que mejoró con el tratamiento con acteósidos. La tinción con HE mostró que la exposición a ELCI dañó la estructura de la corteza y el hipocampo, mientras que el tratamiento con actósidos alivió los cambios estructurales. En comparación con el grupo de control, los niveles de BNDF y CREB en la corteza y el hipocampo del grupo ELCI disminuyeron significativamente. Acteoside revirtió significativamente las expresiones de estas proteínas en ratas ELCI. Mientras tanto, en comparación con el grupo control, los niveles de p-mTOR y p-P70S6K en la corteza y el hipocampo del grupo ELCI aumentaron significativamente, y estos cambios fueron revertidos significativamente ELCI por el acteoside. Sin embargo, el efecto del acteoside sobre la señalización de mTOR fue notablemente bloqueado por la rapamicina, un inhibidor específico de la señalización de mTOR. El acteoside puede atenuar el deterioro de la memoria y mejorar el daño neuronal y la plasticidad sináptica en ratas con depresión, probablemente mediante la inhibición de la vía de señalización mTOR. Acteoside puede servir como un reactivo novedoso para la prevención de la depresión.
Subject(s)
Animals , Rats , Depression/drug therapy , Polyphenols/administration & dosage , Glucosides/administration & dosage , Memory Disorders/drug therapy , Stress, Psychological/complications , Blotting, Western , Fluorescent Antibody Technique , Rats, Sprague-Dawley , Maze Learning , Recognition, Psychology/drug effects , Disease Models, Animal , TOR Serine-Threonine Kinases/antagonists & inhibitors , Polyphenols/therapeutic use , Behavior Rating Scale , MTOR Inhibitors , Glucosides/therapeutic use , Neuronal Plasticity/drug effects , NeuronsABSTRACT
ObjectiveTo reveal the correlation of Rehmannia glutinosa-soil feedback process with the formation of its continuous cropping obstacles through the identification of the root exudates of R. glutinosa and analysis of the specific rhizomicrobes recruited by the root exudate. MethodThe root exudates of R. glutinosa seedlings germinated under sterilized condition and those enriched in the rhizosphere of R. glutinosa cultivated in the field were collected and analyzed using the ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). The highly abundant compounds identified in the root exudates were added into blank soil, and the soil microbial community was profiled using Illumina Miseq sequencing. The bacterial and fungal functions were predicted by PICRUSt and FUNGuild, respectively. ResultThe identification results showed that seven phenylethanoid glycosides were found in R. glutinosa root exudates, and acteoside possessed the highest abundance. In the soil enriched with acteoside, the bacterial genera such as Agromyces, Pseudomonas, Lysobacter, Sphingobium, Pseudoxanthomonas and Sphingomonas were enriched. For the fungi, the genera Neocosmospora, Plectosphaerella and Dactylonectria, and the species such as Neocosmospora rubicola, Plectosphaerella cucumerina, Dactylonectria alcacerensis and Fusarium solani showed higher abundance. The functional analysis indicated the above-mentioned bacterial genera may realize rapid proliferation by utilizing, biodegrading and transforming phenylethanoid glycosides, and some potential fungal pathogens were colonized. ConclusionThe R. glutinsoa-soil feedbacks were likely generated by the phenylethanoid glycosides in the root exudates together with the specific rhizomicrobes. The investigations of R. glutinsoa-soil feedbacks under continuous cropping system are critical to the further understanding of the underlying mechanisms related to its continuous cropping obstacles.
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This study explored the molecular mechanism of acteoside against hepatoma 22(H22) tumor in mice through c-Jun N-terminal kinase(JNK) signaling pathway. H22 cells were subcutaneously inoculated in 50 male BALB/c mice, and then the model mice were classified into model group, low-dose, medium-dose, and high-dose acteoside groups, and cisplatin group. The administration lasted 2 weeks for each group(5 consecutive days/week). The general conditions of mice in each group, such as mental status, diet intake, water intake, activity, and fur were observed. The body weight, tumor volume, tumor weight, and tumor-inhibiting rate were compared before and after administration. Morphological changes of liver cancer tissues were observed based on hematoxylin and eosin(HE) staining, and the expression of phosphorylated(p)-JNK, JNK, B-cell lymphoma-2(Bcl-2), Beclin-1, and light chain 3(LC3) in each tissue was detected by immunohistochemistry and Western blot. qRT-PCR was performed to detect the mRNA expression of JNK, Bcl-2, Beclin-1, and LC3. The general conditions of mice in model and low-dose acteoside groups were poor, while the general conditions of mice in the remaining three groups were improved. The body weight of mice in medium-dose acteoside group, high-dose acteoside group, and cisplatin group was smaller than that in model group(P<0.01). The tumor volume in model group was insignificantly different from that in low-dose acteoside group, and the volume in cisplatin group showed no significant difference from that in high-dose acteoside group. Tumor volume and weight in medium-dose and high-dose acteoside groups and cisplatin group were lower than those in the model group(P<0.001). The tumor-inhibiting rates were 10.72%, 40.32%, 53.79%, and 56.44% in the low-dose, medium-dose, and high-dose acteoside groups and cisplatin group, respectively. HE staining showed gradual decrease in the count of hepatoma cells and increasing sign of cell necrosis in the acteoside and cisplatin groups, and the necrosis was particularly obvious in the high-dose acteoside group and cisplatin group. Immunohistochemical results suggested that the expression of Beclin-1, LC3, p-JNK, and JNK was up-regulated in acteoside and cisplatin groups(P<0.05). The results of immunohistochemistry, Western blot, and qRT-PCR indicated that the expression of Bcl-2 was down-regulated in the medium-dose and high-dose acteoside groups and cisplatin group(P<0.01). Western blot showed that the expression of Beclin-1, LC3, and p-JNK was up-regulated in acteoside and cisplatin groups(P<0.01), and there was no difference in the expression of JNK among groups. qRT-PCR results showed that the levels of Beclin-1 and LC3 mRNA were up-regulated in the acteoside and cisplatin groups(P<0.05), and the level of JNK mRNA was up-regulated in medium-dose and high-dose acteoside groups and cisplatin group(P<0.001). Acteoside promotes apoptosis and autophagy of H22 cells in mice hepatoma cells by up-regulating the JNK signaling pathway, thus inhibiting tumor growth.
Subject(s)
Male , Animals , Mice , Cisplatin/pharmacology , Carcinoma, Hepatocellular/genetics , MAP Kinase Signaling System , Beclin-1 , Apoptosis , Liver Neoplasms/genetics , Necrosis , Proto-Oncogene Proteins c-bcl-2/metabolism , Cell Line, Tumor , RNA, Messenger/metabolism , AutophagyABSTRACT
The study explored the effect of acteoside on renal fibrosis in rats with unilateral ureteral obstruction. Twenty-four healthy adult male SD rats were randomly divided into sham (CON) group, unilateral ureteral obstruction (UUO) group, UUO+acteoside (ACT) group, UUO+benazepril (BZ) group by random number table method. The animal model of renal fibrosis was established by unilateral ureteral ligation. The pathological changes of renal tissues in each group of rats were observed by HE staining and Masson staining. Immunohistochemical method was used to detect the protein expression levels of collagenⅢ, heat shock protein (HSP)-47, connective tissue growth factor (CTGF), α-smooth muscle actin (α-SMA), β-catenin, Bcl-2, and caspase-3. Western blotting was used to detect the protein expression of glucose-regulated protein 78 (GRP78) in renal tissue. Compared with CON group, UUO group had renal tubular dilation and significantly increased interstitial collagen fiber deposition; The degree of renal tubular dilation and interstitial collagen fibers in UUO+ACT group and UUO+BZ group were significantly reduced compared to UUO group, but still significantly heavier than CON group. Compared with CON group, the protein expression levels of collagenⅢ, HSP-47, CTGF, α-SMA, β-catenin and GRP78 in renal tissues of UUO group were significantly higher, while the protein expression level of Bcl-2 was significantly lower (all P<0.05). In comparison with UUO group, the expression levels of Bcl-2 in UUO+ACT group and UUO+BZ group were significantly higher, while the expression levels of other proteins were all significantly lower (all P<0.05). The study shows that acteoside may improve renal fibrosis by decreasing the expression levels of collagenⅢ, HSP-47, CTGF, α-SMA, β-catenin and GRP78, and upregulating the expression level of Bcl-2 in renal tissues.
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Objective To investigate the effect of verbascoside(VB)on endothelial dysfunction in atherosclerotic(AS)rats by regulating high-mobility group protein B1(HMGB1)/receptor for advanced glycation endproducts(RAGE)/nuclear factor κB(NF-κB)signal pathway.Methods The rat model of AS was established by high fat feeding combined with vitamin D3 solution intraperitoneal injection.Rats were divided into the control group(n=10),the model group(n=12),the low(VB-L),medium(VB-M)and high dose(VB-H)VB groups(2,5 and 10 mg/kg,n=10),and the positive control group(simvastatin,5 mg/kg,n=10).The serum level of blood lipids was detected by automatic biochemical analyzer.Pathological changes of aorta were observed by HE staining.Serum levels of inflammatory factors and vascular endothelial cytokines were detected by enzyme-linked immunosorbent assay(ELISA).The level of oxidative stress in rats was detected by micro-method kit.The expression of HMGB1/RAGE signal pathway protein in aorta was detected by Western blot assay.Results Compared with the control group,the intima of aorta in the model group was thickened,plaque appeared in blood vessels,accompanied by lipid deposition and inflammatory cell infiltration.Serum levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),C-reactive protein(CRP),matrix metalloproteinase-9(MMP-9),endothelin-1(ET-1),visfatin,intercellular adhesion molecule-1(ICAM-1)and malondialdehyde(MDA),and HMGB1,RAGE and phosphorylation levels of NF-κB in aorta were obviously increased.Serum levels of high-density lipoprotein cholesterol(HDL-C),nitric oxide(NO),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were obviously decreased(P<0.05).Compared with the model group,pathological changes of rats were obviously improved in the VB-L,VB-M and VB-H groups and the simvastatin group.Serum levels of TC,TG,LDL-C,TNF-α,IL-1β,CRP,MMP-9,ET-1,visfatin,ICAM-1,MDA,and HMGB1,RAGE,phosphorylation levels of NF-κB in aorta were obviously decreased,and serum levels of HDL-C,NO,SOD and GSH-Px were obviously increased(P<0.05).Conclusion VB can down-regulate the expression of HMGB1/RAGE/NF-κB signal pathway protein,inhibit inflammation and oxidative stress in AS rats,and improve lipid metabolism and vascular endothelial function.
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The Compound Cheqian Tablets are derived from Cheqian Power in Comprehensive Recording of Divine Assistance, and they are made by modern technology with the combination of Plantago asiatica and Coptis chinensis. To investigate the material basis of Compound Cheqian Tablets in the treatment of diabetic nephropathy, in this study, the chemical components of Compound Cheqian Tablets were characterized and analyzed by UPLC-Q-TOF-MS/MS, and a total of 48 chemical components were identified. The identified chemical compounds were analyzed by network pharmacology. By validating with previous literature, six bioactive compounds including acteoside, isoacteoside, coptisine, magnoflorine, palmatine, and berberine were confirmed as the index components for qua-lity evaluation. Furthermore, the content of the six components in the Compound Cheqian Tablets was determined by the "double external standards" quantitative analysis of multi-components by single marker(QAMS), and the relative correction factor of isoacteoside was calculated as 1.118 by using acteoside as the control; the relative correction factors of magnoflorine, palmatine, and berberine were calculated as 0.729, 1.065, and 1.126, respectively, by using coptisine as the control, indicating that the established method had excellent stability under different conditions. The results obtained by the "double external standards" QAMS approximated those obtained by the external standard method. This study qualitatively characterized the chemical components in the Compound Cheqian Tablets by applying UPLC-Q-TOF-MS/MS and screened the pharmacodynamic substance basis for the treatment of diabetic nephropathy via network pharmacology, and primary pharmacodynamic substance groups were quantitatively analyzed by the "double external stan-dards" QAMS method, which provided a scientific basis for clarifying the pharmacodynamic substance basis and quality control of Compound Cheqian Tablets.
Subject(s)
Humans , Tandem Mass Spectrometry , Berberine/pharmacology , Chromatography, High Pressure Liquid/methods , Network Pharmacology , Diabetic Nephropathies , Drugs, Chinese Herbal/chemistry , Quality Control , TabletsABSTRACT
This study aims to reveal the endogenous metabolic characteristics of acteoside in the young rat model of purinomycin aminonucleoside nephropathy(PAN) by non-targeted urine metabolomics and decipher the potential mechanism of action. Biochemical indicators in the urine of rats from each group were determined by an automatic biochemical analyzer. The potential biomarkers and related core metabolic pathways were identified by ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). MetaboAnalyst 5.0 was used to establish the receiver operating characteristic(ROC) curve for evaluating the clinical diagnostic performance of core metabolites. The results showed that acteoside significantly decreased urinary protein-to-creatinine ratio in PAN young rats. A total of 17 differential metabolites were screened out by non-targeted urine metabolomics in PAN young rats and they were involved in phenylalanine metabolism and phenylalanine, tyrosine and tryptophan biosynthesis. Thirtten differential metabolites were screened by acteoside intervention in PAN young rats, and they were involved in phenylalanine metabolism and arginine and proline metabolism. Among them, leucylproline and acetophenone were the differential metabolites that were significantly recovered after acteoside treatment. These pathways suggest that acteoside treats PAN in young rats by regulating amino acid metabolism. The area under the curve of two core biomarkers, leucylproline and acetophenone, were both greater than 0.9. In summary, acteoside may restore amino acid metabolism by regulating endogenous differential metabolites in PAN young rats, which will help to clarify the mechanism of acteoside in treating chronic glomerulonephritis in children. The characteristic biomarkers screened out have a high diagnostic value for evaluating the treatment of chronic glomerulonephritis in children with acteoside.
Subject(s)
Humans , Child , Rats , Animals , Puromycin Aminonucleoside , Metabolomics/methods , Biomarkers/urine , Chromatography, High Pressure Liquid/methods , Acetophenones , Glomerulonephritis , Phenylalanine , Amino AcidsABSTRACT
Objective To investigate the effect and mechanism of acteoside (ACT) in inhibiting epithelial-mesenchymal transition (EMT) in human hepatoma HCCLM3 cells by regulating the ERK1/2 pathway. Methods CCK-8 assay was used to detect the effect of hepatocellular carcinoma cell proliferation. The invasion and migration of HCC cells were detected by scratch and Transwell tests. The mRNA and protein expression levels of the ERK1/2 signaling pathway and EMT-related genes (E-cadherin and N-cadherin) were detected by real-time PCR and Western blot analyses. Results ACT reduced the activity of HCCLM3 cells and inhibited the proliferation of HCC cells, and the effects had certain correlation with drug concentration and time. ACT inhibited the migration and invasion process of HCCLM3 cells in a concentration-dependent manner. ACT downregulated the mRNA and protein expression of genes related to the ERK1/2 signaling pathway. It increased the mRNA and protein expression levels of the EMT-related gene E-cadherin but decreased those of N-cadherin. Conclusion ACT could inhibit EMT and the invasion and migration of HCCLM3 cells in human hepatoma, and the underlying mechanism is closely related to the downregulation of the ERK1/2 signaling pathway.
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italic>Rehmannia glutinosa belongs to the Scrophulariaceae family with important medicinal value. In order to effectively explore the transcriptome information of R. glutinosa and identify the genes encoding enzymes involved in phenylethanol glycoside (PhGs) biosynthesis, the leaves, stems and tuberous roots of R. glutinosa were used for transcriptome sequencing using Pacific Biosiences RS II platform. A total of 27 773 transcripts were generated with an average length of 2 380 bp, and 27 236 coding sequences (CDS) were predicted. Using BLAST software, non-redundant transcript sequences were annotated with NR, NT, GO, COG, KEGG, SwissProt and Interpro databases and a total of 27 399 annotated genes were obtained. Among them, the number of genes related to Sesamum indicum in the NR database was the highest (81.44%), which is consistent with their evolutionary relationship. Enzymes likely involved in the biosynthesis of isoacteoside, echinacoside, cistanosides A, cistanosides F, 2′-acetylacteoside and leonoside F were identified, and 143 genes were identified in R. glutinosa full-length transcriptome. The expression levels of 19 genes correlated with acteoside content in twelve tissues of R. glutinosa, and most showed higher expression levels in leaf tissues and floral organs. This study provides more reliable transcriptome data for screening R. glutinosa for functional genes and provides a foundation for the study of the molecular mechanisms of PhGs biosynthesis.
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OBJECTIVE To inv estigate the in vitro inhibitory effects of acteoside on cytochrome P 450(CYP)enzymes in liver microsomes of rats. METHODS Using probe substrates method ,acteoside(0.1,0.3,1,3,10,30 μmol/L)was incubated with probe substrates phenacetin ,mephentoin,diclofenac,coumarin,dextromethorphan and testosterone (substrates of CYP 1A2, CYP2C19,CYP2C9,CYP2A6,CYP2D6 and CYP 3A4 enzymes,respectively)in liver microsomes of rats. Another blank control group and positive inhibitor group [ α-naphthoflavone,ticlopidine,sulfabendazole,pilocarpine,quinidine and ketoconazole (inhibitors of CYP 1A2,CYP2C19,CYP2C9,CYP2A6,CYP2D6 and CYP 3A4 enzymes,respectively)] were set up. Using indapamide as the internal standard , the contents of corresponding metabolites (acetaminophen, 4-hydroxymephenytoin, 4-hydroxydiclofenac,7-hydroxycoumarin,dextran,6 β-hydroxytestosterone) were detected by ultra high performance liquid chromatography-tandem mass spectrometry . The IC 50 values were calculated by GraphPad v 8.0 software. By computer molecular docking technology ,acteoside and positive inhibitors were molecularly docked with the CYP enzyme ,and the binding mode and strength of the two molecules were analyzed. RESULTS The IC 50 values of acteoside to CYP 1A2 and CYP 2A6 enzymes were more than 30 μmol/L,and those of acteoside to CYP 2D6,CYP2C19,CYP2C9 and CYP 3A4 enzymes were 24.87,21.52,12.56 and 7.55 μmol/L,respectively. The hydrogen bond and hydrophobic force could form between acteoside and CYP 3A4 enzyme,and the hydrogen bond and electrostatic interaction could form between ketoconazole and CYP 3A4 enzyme. The binding free energy of acteoside and ketoconazole to CYP 3A4 enzyme were - 10.2 and - 12.4 kcal/mol (1 kcal/mol=4.19 kJ),respectively. CONCLUSIONS Acteoside shows moderate inhibitory effect on CYP 3A4 enzyme in liver microsomes of rats ,and its affinity is equivalent to that of positive inhibitor ;the compound shows weak inhibitory effect on other 5 CYP enzymes.
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OBJECTIVE To explore the effects of acteoside on hypoxia/reoxygena tion(H/R)-induced cardiomyocyte damage by regulating Rho family GTPase 3(Rnd3)/nuclear factor κB(NF-κB)pathway. METHODS The H 9c2 cardiomyocyte were divided into control group (no administration ,no modeling ),H/R group (only modeling ),H/R+AS-L group ,H/R+AS-M group , H/R+AS-H group (10,30,90 μmol/L acteoside for above 3 groups firstly ,and then modeling ),H/R+pcDNA group [transfecting pcDNA (empty vector ) firstly,and then modeling] ,H/R + pcDNA-Rnd 3 group [overexpression of Rnd 3 by transfecting pcDNA-Rnd3(Rnd3 overexpression vector )firstly,and then modeling] ,H/R+AS-H+si-NC group [transfecting si-NC (negative control)firstly,and then giving 90 μmol/L acteoside and modeling],H/R+AS-H+si-Rnd3 group [inhibiting overexpression of Rnd 3 by transfecting si-Rnd 3 (Rnd3 small interfering RNA ) firstly,and then giving 90 μ mol/L acteoside and modeling]. After corresponding treatment ,the apoptotic rate ,release of lactate dehydrogenase (LDH),malondialdehyde(MDA)level,the activity of superoxide dismutase (SOD),the level of tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β)and interleukin- 6(IL-6), mRNA and protein expression of Rnd 3 and NF-κB subunit p65(NF-κB p65),the expression of aspartate proteolytic enzyme 3 (Cleaved Caspase- 3)protein and Cleaved Caspase- 9 protein were detected. RESULTS Different concentrations of acteoside could reduce the apoptotic rate of H/R-induced H 9c2 cardiomyocyte,the protein expressions of Cleaved Caspase- 3 and Cleaved Caspase-9,mRNA and protein expressions of NF-κB p65,the levels of LDH release and MDA ,TNF-α,IL-1β and IL-6,while increase the activity of SOD and mRNA and protein expressions of Rnd 3(P<0.05),in a dose-dependent manner. Overexpression of Rnd 3 could decrease the apoptotic rate of H 9c2 cardiomyocyte,protein expressions of NF-κB p65,Cleaved Caspase- 3 and Cleaved Caspase- 9, the levels of LDH release , MDA, TNF-α,IL-1β and IL-6,while increase the protein expression of Rnd 3 and the activity of SOD (P<0.05). The inhibition overexpression of Rnd 3 could weaken the inhibitory effects of acteoside on H/R-induced apoptosis of H 9c2 cardiomyocyte, oxidative stress and inflammatory reaction (P<0.05). CONCLUSIONS Acteoside could regulate Rnd 3/NF-κ B pathway by promoting the expression of Rnd 3 and inhibiting the expression of NF-κB p65,inhibit cardiomyocyte apoptosis ,oxidative stress and inflammation reaction so as to relieve the H/R-induced cardiomyocyte damage.
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Aim To investigate the effects of acteoside (AS) on BRAIN AKT and NFkB in APP/PS1 double transgenic mice. Methods Fifty healthy APP/PS1 transgenic mice, half male and half female, and 10 control C57 mice were given the drug by gavage for 60 consecutive days. During the period of administration, and new object recognition experiments were conducted to detect the expression of AKT and NFkB related proteins in the hippocampus and cortex of the mice. Results Compared with model group, AS could significantly improve the nesting behavior of mice and en- hance the interest of exploring new objects in mice with Alzheimer' s disease. At the same time, compared with model group, AS significantly reduced the ratio of NFkB p-p65 /NFkB p65 in hippocampus and cortical tissues, and increased the ratio of p-AKT-308 /AKT and p-AKT473 /AKT. Conclusions AS may inhibit the apoptosis of nerve cells and protect nerve cells through the regulation of AKT and NFkB to treat neurodegenerative diseases.
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Acteoside is among the most widespread of thedisaccharide caffeoyl esters that are widely distributed in the plant kingdom with diverse biological activities. Recent studies have shown that acteoside has neuroprotective activity in neurodegenerative diseases. This review examines and extrapolates from the recent literature to build support for the use of acteoside in mitigating neuropathy in neurodegenerative disease, including Parkinson ' s disease (PD) and Alzheimer' s disease (AD). We summarize the main pharmacokinetic parameters of acteoside in animals after different administration routes. Meanwhile, we point out both problems and shortcomings, and highlight its future development trend.
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OBJECTIVE:To estab lish a me thod for simultaneous determination of morroniside ,loganin,echinacoside and acteoside in Huanshao capsules. METHODS :HPLC method was adopted. The determination was performed on Zhongpuhong RD-C18 column with mobile phase consisted of acetonitrile- 0.1% formic acid solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm (morroniside,loganin) and 330 nm (echinacoside,acteoside). The column temperature was set at 35 ℃,and sample size was 10 μL. RESULTS:The linear range were 5.29-105.80 μg/mL for morroniside, 4.49-89.88 mg/L for loganin ,16.26-325.25 mg/L for echinacoside and 16.31-326.25 mg/L for acteoside ,r values were 0.999 9. RSDs of precision ,stability (24 h),reproducibility and durability tests were all lower than 2.0% . The recoveries were 94.34% -96.23%(RSD=0.81% ,n=6),97.04% -98.89%(RSD=0.73% ,n=6),96.23% -98.08%(RSD=0.82% ,n=6), 95.40%-98.47%(RSD=1.23%,n=6),respectively. The contents of above 4 components in 11 batches of Huanshao Capsules were 0.190-0.704,0.439-0.857,2.723-4.475 and 0.589-1.035 mg/g,respectively. CONCLUSIONS :Established method is specific , precise and can be used for content determination of 4 components in Huanshao capsules.
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Objective: To establish HPLC fingerprints of Qiwei Tangmaishu tablet and simultaneously determine the contents of the six constituents, acteoside, martynoside, calycosin 7-O-β- D-glucopyranoside, schisandrin, formononetin and deoxyschizan- drin. Methods: The analysis of 50% methanol extract of this drug was performed on a 30℃ thermostatic Waters Symmetry C18 column, with the mobile phase comprising of acetonitrile(A)-0.2% formic acid solution(B)flowing at 1.0 ml/min in a gradient elution manner. The detection wavelength was set at 330 nm for acteoside and martynoside, 254 nm for calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin. Results: There were thirteen common peaks in the fingerprints of ten batches of samples with the similarities more than 0.9. Acteoside, martynoside, calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin showed good linear relationship within the ranges of 1.47-36.75, 0.66-16.50, 0.89-22.25, 2.58-64.50, 1.91-47.75 and 0.77-19.25 μg/ml(r≥0.9993), and their average recoveries were 97.57%, 99.22%, 96.69%, 100.01%, 98.79% and 96.77%, with the RSD of 0.79%, 1.54%, 0.61%, 0.64%, 0.83% and 0.36%, respectively. Conclusion: The established method is easily operable and repeatable, which could be used for quality control of Qiwei Tangmaishu tablet.
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Objective: To study the content variation and chemical composition of Siwu Decoction between mixed decoction and single decoction comprehensively, and then explore variation rule of Siwu Decoction by different decocting methods based on material basis. Methods: Components of Siwu Decoction were identified by LC-MS/MS and an UPLC wavelength switching method for simultaneously determining the contents of multiple compounds in Siwu Decoction was established based on the idea of TCM chemistry holography. The mixed and single decoction samples were prepared and tested. Experimental data was compared to analyze material basis differences and variation rule of Siwu Decoction by different decocting methods. Results: A total of 72 compounds were identified and assigned, 18 compounds were quantitative detected and all of 18 analytes showed good linearity (R2 ≥ 0.999) within the test range. The relative standard deviations of the precision, repeatability and stability were not exceeding 2.0%, and the recoveries were in the range of 97%-105%. Analysis of Siwu Decoction samples showed dissolution of ligustilide, 3-n- butylphthalide, catechin, gallic acid and paeoniflorin was affected by the change of solvent volume and dissolution of aucubin, catechin, oxypaeoniflorin, paeoniflorin and acteoside were higher in mixed decoction than single decoction obviously. Compared to single decoction, the kinds of compounds in mixed decoction did not change significantly but the content showed notable variety. Conclusion: Through the study of chemistry holography, the composition and content of compounds in TCM mixed decoction and herbs single decoction can be compared and analyzed comprehensively to provide a new perspective for the study on the rule of TCM decoction and dissolution. TCM chemistry holography study may become a useful exploration of the TCM quality study.
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Objective: To clone the acteoside synthase gene (RgAcS1) from Rehmannia glutinosa, and analyze its subcellular localization and expression pattern. Methods: The cDNA sequence of RgAcS1 was identified based on the annotation of the transcriptome data of R. glutinosa, and the RgAcS1 gene was cloned by polymerase chain reaction (PCR). Constructing the GFP fusion expression vector and observing the subcellular localization of RgAcS1 mediated by Agrobacterium tumefaciens. The expression pattern of RgAcS1 in different parts of tuberous root of R. glutinosa was detected by real-time fluorescence quantitative PCR (qRT-PCR). Results: A full-length coding sequence of a shikimate-O-hydroxy cinnamoyl transferase from R. glutinosa was obtained and named RgAcS1. The length of the RgAcS1 cDNA was 1659 bp, including an open reading frame (ORF) of 1 296 bp, encoding 431 amino acid residues, the molecular weight of the protein was 475 900, and it has a typical domain of shikimic acid-O-hydroxy cinnamoyl transferase. The result of subcellular localization showed that RgAcS1 was mainly distributed in cytoplasm and also in nucleus. The qRT-PCR analysis showed that the expression levels of RgAcS1 were higher in the periderm and root hair of R. glutinosa tuberous root, but lower in the xylem and phloem. The expression levels of RgAcS1 were higher in non-radial striation than that in radial striation of BJ1, QH1 and 85-5. Conclusion: In this study, we obtained the cDNA sequence of RgAcS1, and analyzed the subcellular location and expression patterns of RgAcS1, which will lay foundations for further study on roles of RgAcS1 gene in the synthesis of acteoside in R. glutinosa.
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Aim To investigate the effect of acteoside on the changes of immune function and aging process in SAMP8 mice via detecting the proportion of immune cells and the changes of immune factors. Methods Sixty male healthy SAMP8 mice and twelve SAMR1 mice were randomly divided into six groups. They were administered by gavage once a day for 90 days. Aging scoring test was conducted regularly. The immune cells and immune factors in plasma and liver were detected. Results Compared with model group, acteoside could increase CD4+lymphocytes in SAMP8 mice and effectively inhibit the increase of CD8+lymphocytes; it played a greater role in spleen than in peripheral blood. At the same time, Thl/Th2 was significantly reduced in model group, and the ratio of Thl/Th2 factor increased in low and medium dose group. Conclusions Acteoside can increase the number of helper lymphocytes, effectively inhibit the increase of killer lymphocytes, and regulate the dynamic balance of Th1 and Th2 type immune inflammatory factors in SAMP8 mice, so as to improve the immune function of the body and delay the aging process of the body.
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Objective: To provide the theoretical basis for determining the best harvesting plant organ and harvesting period, and investigate the content of chemical constituents of Callicarpa nudiflora in different plant organs and different growth periods. Methods: The contents of total flavonoids, total phenolic acid and total saponins were determined by ultraviolet spectrophotometry, and the seven components were determined by HPLC. The ANOVA and PCA methods were used to analyze the content of each constituent. Results: The dry extract rate, the contents of total flavonoids, total phenolic acid, total saponins, forsythiaside B, acteoside, isoacteoside, and apigenin-7-O-β-D-glucopyranoside in functional leaves were the highest, while the contents of caffeic acid, galuteolin and luteolin in tender leaves were the highest, and all of them were significantly different from the young shoots (P 0.05). The contents of total phenolic acid, total saponins, forsythiaside B, and acteoside were the highest in the FB period, and there was no significant difference with the EFS period (P > 0.05). The contents of galuteolin and apigenin-7-O-β-D-glucopyranoside were the highest in the earlier fruit maturation (EFM) period and the later fruit maturation (LFM) period, respectively, and there was no significant difference with the EFS period (P > 0.05). The contents of each chemical component were reduced to the minimum at the fruit-drop (FD) period, and it was significantly different from that at the EFS period and the FB period (P < 0.05). According to the comprehensive evaluation model constructed by PCA, the comprehensive score of the EFS period was the highest (F = 3.252), followed by the FB period (F = 3.011). Conclusion: Main chemical constituents of C. nudiflora were significantly different in harvesting parts and growth periods. The contents of main chemical constituents were higher in functional leaves and tender leaves, and the contents of main chemical constituents were higher from FB period to EFS period.
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Objective: To optimize the water extraction process of Siwu Decoction by BP neural network combined with orthogonal experiment. Methods: The water amount, the extraction time, and the extraction times were taken as factors. Entropy weight method was used to calculate the comprehensive scores of the multi-indicators of eight active components of 5-hydroxymethylfurfural, chlorogenic acid, caffeic acid, paeoniflorin, ferulic acid, verbascoside, senkyunolide A, and ligustilide in R language environment. Using comprehensive score as an evaluation indicator, the BP neural network model was established by orthogonal experiment design, and the optimal water extraction process of Siwu Decoction was predicted through network training. Results: The optimized extraction process of Siwu Decoction was carried out by adding 8 times of water and extracting 3 times for 1 h. The relative error between the network predicted value and the actual measured value of the test sample was less than 1%. Conclusion: The established mathematical model can analyze and predict the water extraction process of Siwu Decoction. The obtained process is stable and feasible, and can effectively extract the active ingredients in Siwu Decoction.