ABSTRACT
BACKGROUND: Herba Epimedii, a commonly used traditional herb, has been proven effective in ameliorating osteoporosis. However, the active ingredients and potential mechanism need further exploration. OBJECTIVE: To screen active ingredients of Herba Epimedii with the effect of ameliorating osteoporosis and to explore their potential mechanisms. METHODS: TCMSP and Swiss Target Prediction were applied to collect the ingredients of Herba Epimedii and their targets. UniProt, GeneCards, TTD, DisGeNET, and OMIM were adopted to search osteoporosis-related genes. STRING and DAVID were used to perform enrichment analysis. Effects of screened ingredients were evaluated on MC3T3-E1 cells and RAW264.7 cells, respectively. RESULTS: Eleven ingredients were screened by Network Pharmacology. They exerted a promoting effect on MC3T3-E1 cells (10-9-10-5 M). The ingredients didn't significantly affect ALP activity and osteoblastogenesis-related genes. Baohuoside 1, Sagittatoside B, Chlorogenic acid, Cryptochlorogenic acid, and Neochlorogenic acid significantly increased calcium depositions. The ingredients didn't exhibit a dose-dependent inhibition or promotion on RAW264.7 cells. Baohuoside 1, Sagittatoside B, Neochlorogenic acid, Cryptochlorogenic acid, Icariin, Epimedin A, Chlorogenic acid, Sagittatoside A, and Epimedin C suppressed the level of TRACP. Baohuoside 1, Sagittatoside B, Cryptochlorogenic acid, Neochlorogenic acid, Chlorogenic acid, Sagittatoside A, and Icariin decreased the number of multinucleated osteoclastic cells. Baohuoside 1, Sagittatoside B, and Cryptochlorogenic acid could significantly inhibit MMP-9 expression. CONCLUSION: Neochlorogenic acid, Sagittatoside B, Chlorogenic acid, and Cryptochlorogenic acid promoted MC3T3-E1 differentiation, among which Neochlorogenic acid showed significant promotion in viability, mineralization, and OPN expression. Baohuoside 1, Sagittatoside B, Cryptochlorogenic acid, Neochlorogenic acid, Chlorogenic acid, and Icariin inhibited RAW264.7 differentiation, among which Baohuoside 1 showed significant inhibition on TRACP, multinucleated osteoclastic cells number and MPP-9 expression. The mechanism might relate to the FoxO signaling pathway, MAPK signaling pathway, and TNF signaling pathway.
ABSTRACT
Periodic pillars of semiconductor in sub-wavelength size can serve multiple roles as diffracting, trapping and absorbing light for effective photoelectric conversion which has been intensively studied in the visible range. Here, we design and fabricate the micro-pillar arrays of AlGaAs/GaAs multi quantum wells(QWs) for high performance detection of long wavelength infrared light. Compared to its planar counterpart, the array offers 5.1 times intensified absorption at peak wavelength of 8.7 µm with 4 times shrinked electrical area. It's illustrated by simulation that the normal incident light is guided in the pillars by HE11 resonant cavity mode to form strengthened Ez electrical field, which enables the inter-subband transition of n-type QWs. Moreover, the thick active region of dielectric cavity that contains 50 periods of QWs with fairly low doping concentration will be beneficial to the optical and electrical merits of the detectors. This study demonstrates an inclusive scheme to substantially raise the signal to ratio of infrared detection with all-semiconductor photonic structures.
ABSTRACT
ß-Elemonic acid is one of the main active ingredients isolated from Boswellia carterii Birdw. which has been reported to exhibit potential anti-inflammatory and anti-cancer activities. There is few information about pharmacokinetics and tissue distribution of ß-elemonic acid by now. In this study, an ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-MS/MS) method has been developed and validated to determine ß-elemonic acid in rat plasma and various tissues after intragastric administration. Oleanolic acid was chosen as an internal standard (IS) and the plasma/tissue samples were pretreated with one-step liquid-liquid extraction. Chromatographic separation was accomplished on Eclipse Plus C18 analytical column (2.1 × 50 mm, 1.8 µm) utilizing a gradient mobile phase system consisting of water (with 0.1% ammonia-solution) and acetonitrile. ß-Elemonic acid and IS were detected and quantified using negative electrospray ionization in multiple reaction monitoring (MRM) mode with transitions of m/z 453.3 â 423.5 for ß-elemonic acid and m/z 455.3 â 407.6 for IS. ß-Elemonic acid showed good linearity over the investigated concentration range (r > 0.9934) in rat plasma and tissue sample. The method was successfully applied for determination of ß-elemonic acid in bio-samples. A bimodal phenomenon appeared in the plasma concentration-time curve of the ß-elemonic acid. The highest tissue concentrations were found in the intestine including jejunum, ileum and colon.
Subject(s)
Chromatography, High Pressure Liquid/methods , Tandem Mass Spectrometry/methods , Triterpenes/blood , Triterpenes/pharmacokinetics , Animals , Linear Models , Male , Rats , Reproducibility of Results , Sensitivity and Specificity , Tissue Distribution , Triterpenes/chemistryABSTRACT
The human Immunodeficiency Virus Transactivator (TAT) protein transduction peptide is a trans-transcription activator encoded by HIV-1. It is rich in basic amino acids, and capable of efficiently mediating the passage of exogenous macromolecules through a variety of membrane structures, such as the cytoplasmic membrane and the blood-brain barrier. Metallothionein (MT) is a protein with low molecular weights and rich cysteine contents. It plays important roles in maintaining the dynamic balance of metal contents in the body, in the detoxification of heavy metals and in defense against oxidative stress. Based on the full-length MT cDNA previously cloned from Sinopotamon henanense, we aim to prepare a TAT-mediated recombinant fusion protein that can cross the membrane and enter the cell by means of genetic engineering. The hydroxyl radical scavenging rate and total antioxidant capacity of TAT-MT were measured in vitro. An immunofluorescence technique was used to detect the transmembrane activity. An MTT assay was used to study the repair effect of H
ABSTRACT
Melanoma associated antigen family A1 (MAGEA1) is expressed in germ cells and tumors of various histological origins, but its mechanism is still unclear. In this study, the eukaryotic recombinant MAGEA1 expression plasmids with Flag or GFP tags were constructed and transfected into HeLa and HEK293T cells. Western blotting, immunocytochemistry, co-immunoprecipitation, nuclear protein and cytoplasmic protein separation, and mitochondrial isolation were used to detect the expression and location of MAGEA1 and its interaction with other proteins in cells. The results of immunocytochemistry (ICC) and Western blotting showed that the overexpressed MAGEA1 was mainly localized in the cytoplasm and partially co-localized with mitochondria. Co-immunoprecipitation experiments verified the interactions between MAGEA1 and TRIM31, SNW1, HDAC1, and found that MAGEA1 may mainly interact with HDAC1 in the cytoplasm. The studies above indicate that MAGEA1 may be involved in different cellular biological processes and co-localize with mitochondria. It interacts with TRIM31, SNW1 and HDAC1, while MAGEA1 may mainly interact with HDAC1 in the cytoplasm. We propose that it may be involved in protein ubiquitination and the Notch signaling pathway. The results of this study laid an experimental foundation for the subsequent in-depth study of the mechanism of MAGEA1.
ABSTRACT
A method for the simultaneous quantification of 13 bioactive compounds (psoralen, isopsoralen, isobavachin, bakuchalcone, neobabaisoflavone, bavachin, corylin, psoralidin, isobavachalcone, bavachinin, corylifol A, bavachalcone, and bakuchiol) by ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry has been developed and validated in rat plasma. Osthol was used as an internal standard and plasma samples were pretreated with one-step liquid-liquid extraction. These analytes were separated using a gradient mobile phase system of water and acetonitrile at a flow rate of 0.2 mL/min on a reverse-phase C18 column and analyzed in the selected multiple reactions monitoring mode. All calibration curves were linear (r > 0.9952) over the tested ranges. The intra- and interday accuracy and precisions of these analytes at three different concentration levels were within the acceptable limits of <15% at all concentrations. The mean recoveries of these analytes at three concentrations were more than 60.2% and the matrix effects were in the range of 85-115%. Stability studies proved that the analytes were stable under the tested conditions. The developed method was applied to evaluating the pharmacokinetic study of 13 bioactive compounds after oral administration of Psoraleae Fructus in rat of different genders. Some active compounds in Psoraleae Fructus had sex-related pharmacokinetics.
Subject(s)
Psoralea/chemistry , Animals , Benzofurans/blood , Benzofurans/pharmacokinetics , Chalcones/blood , Chalcones/pharmacokinetics , Chromatography, High Pressure Liquid , Coumarins/blood , Coumarins/pharmacokinetics , Female , Ficusin/blood , Ficusin/pharmacokinetics , Flavones/blood , Flavones/pharmacokinetics , Flavonoids/blood , Flavonoids/pharmacokinetics , Furocoumarins/blood , Furocoumarins/pharmacokinetics , Male , Mass Spectrometry , Molecular Structure , Phenols/blood , Phenols/pharmacokinetics , Rats , Rats, Sprague-DawleyABSTRACT
Psoralen is a furanocoumarin compound found in many herb medicines and is claimed to contribute to the hepatotoxicity caused by lots of traditional Chinese medicine. So far, there has been no research on the differences in pharmacokinetics of single and repeated dosing of psoralen. Moreover, the research on the cumulative toxicity of low concentration and long-term administration on cells has not been reported. Therefore, this study investigated the pharmacokinetic differences and the accumulated cytotoxicity of psoralen from repeated administration. The study found that after single or repeated administration of psoralen for 3 months at various dosages (14, 28, and 56 mg/kg), the pharmacokinetic parameters of female rats between single dose and repeated dose administration are totally different. Compared with a single administration, multiple administrations increased psoralen's in vivo exposure, prolonged the peak time, prolonged the half-life of the drug, reduced the drug clearance rate, and prolonged the drug's stay in the body. HepG2 cells were exposed to low doses (5, 10, 20, or 40 µM) of psoralen for 1, 2, 3, or 4 days. A 20 and 40 µM dose of psoralen did not induced cell death in the 1st day but significantly decreased the cell viability at the 3rd and 4th day of repeated administration, respectively. In addition, multiple administrations of psoralen decreased cell viability due to G2 arrest.
ABSTRACT
Rhodiola rosea L., a worldwide botanical adaptogen, has been confirmed to possess protective effects of inflammatory injury for many diseases, including cardiovascular diseases, neurodegenerative diseases, diabetes, sepsis, and cancer. This paper is to review the recent clinical and experimental researches about the anti-inflammatory effects and the related mechanisms of Rhodiola rosea L. extracts, preparations, and the active compounds. From the collected information reviewed, this paper will provide the theoretical basis for its clinical application, and provide the evidences or guidance for future studies and medicinal exploitations of Rhodiola rosea L.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Plant Extracts/pharmacology , Rhodiola/chemistry , Animals , Anti-Inflammatory Agents/isolation & purification , Humans , Inflammation/drug therapy , Inflammation/pathologyABSTRACT
BACKGROUND: The main bioactive components of Fructus psoraleae, such as psoralen and isopsoralen, are known to be hepatotoxic. However, its underlying mechanism is to be elucidated. METHODS: To address this, SD rats were randomly divided into control group, 60 mg/kg psoralen group and 60 mg/kg isopsoralen group. Blood was collected to detect serum biochemical indices. RNA was extracted from liver samples, and then, cDNA gene expression profiles were analysed. RESULTS: Psoralen administration significantly up-regulated serum AST (aspartate aminotransferase) while addition of isopsoralen increased serum ALT (alanine aminotransferase), AST, TBA (total bile acid) and TG (total triglyceride) levels. A total of 172 differentially expressed genes (DEGs) were acquired between psoralen group and control group while 884 DEGs were screened between isopsoralen group and control group. Chemical Carcinogenesis and Metabolism of Xenobiotics by Cytochrome P450 were the two most significantly enriched pathways as revealed by DEGs. Liver was the most impacted organ, and endoplasmic reticulum was the most impacted organelle in subcellular level. Finally, some kinds of cancers and cytochrome p450 oxidoreductase deficiency were predicted. Taken together, psoralen and isopsoralen might cause hepatotoxicity mainly through cytochrome P450 metabolism of xenobiotics. Furthermore, Cyp1a1, Cyp1a2, Gstm1 and Akr7a3 worked as key genes in hepatotoxicity. Moreover, endoplasmic reticulum was the main target subcellular structure in hepatotoxicity induced by psoralen and isopsoralen.
Subject(s)
Chemical and Drug Induced Liver Injury/etiology , Ficusin/toxicity , Furocoumarins/toxicity , Liver/drug effects , Animals , Chemical and Drug Induced Liver Injury/genetics , Chemical and Drug Induced Liver Injury/metabolism , Gene Expression Profiling , Liver/metabolism , RNA, Messenger , Random Allocation , Rats , Rats, Sprague-DawleyABSTRACT
The purpose of this research was to extract and separate the compounds from frankincense, and then evaluate their anti-inflammatory effects. The isolated compound was a representative tetracyclic triterpenes of glycine structure according to ¹H-NMR and 13C-NMR spectra, which is ß-elemonic acid (ß-EA). We determined the content of six different localities of frankincense; the average content of ß-EA was 41.96 mg/g. The toxic effects of ß-EA administration (400, 200, 100 mg/kg) for four weeks in Kunming (KM) mice were observed. Compared with the control group, the body weight of mice, the visceral coefficients and serum indicators in the ß-EA groups showed no systematic variations. The anti-inflammatory effects of ß-EA were evaluated in LPS-induced RAW264.7 cells, xylene-induced induced ear inflammation in mice, carrageenin-induced paw edema in mice, and cotton pellet induced granuloma formation in rats. ß-EA inhibited overproduction of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6), monocyte chemotactic protein 1 (MCP-1), soluble TNF receptor 1 (sTNF R1), Eotaxin-2, Interleukin 10 (IL-10) and granulocyte colony-stimulating factor (GCSF) in the RAW264.7 cells. Intragastric administration with ß-EA (300, 200, and 100 mg/kg in mice, and 210, 140, and 70 mg/kg in rats) all produced distinct anti-inflammatory effects in vivo in a dose-dependent manner. Following treatment with ß-EA (300 mg/kg, i.g.), the NO level in mice ears and PGE2 in mice paws both decreased (p < 0.01). In conclusion, our study indicates that ß-EA could be a potential anti-inflammatory agent for the treatment of inflammatory diseases.
Subject(s)
Anti-Inflammatory Agents/administration & dosage , Dinoprostone/metabolism , Frankincense/chemistry , Inflammation/drug therapy , Triterpenes/administration & dosage , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Carrageenan/adverse effects , Cell Survival/drug effects , Dose-Response Relationship, Drug , In Vitro Techniques , Inflammation/chemically induced , Lipopolysaccharides/adverse effects , Mice , Nitric Oxide/metabolism , RAW 264.7 Cells , Rats , Triterpenes/chemistry , Triterpenes/pharmacology , Xylenes/adverse effectsABSTRACT
OBJECTIVE: To study the effect of toosendanin (TSN)on the apoptosis of human ovarian cancer cell and to clarify the related mechanism. METHODS: CAVO-3 and A2870 cells were treated with toosendanin of different concentrations, and CCK-8 method was used to detect the cell survival rate, colorimetric method was used to measure the activities of Caspase-3 and Caspase-9, EdU method was used to determine the cell proliferation rate, and Western blot method was used to test the expressions of Bcl-2, Bax and Cyto-C protein. RESULTS: TSN could significantly inhibit the survival of CAVO-3 and A2870 cells with dose-(r=0.869 1, P<0.05) and time-(r=0.776 5, P<0.05) dependent manners. The survival rate of A2870 cell with TSN dropped significantly (P<0.05). The activities of Caspase-3 and Caspase-9 were significantly increased by TSN in CAVO-3 and A2870 cells, however, the inhibitors of Caspase-3 (z-DEVE-FMK) and Caspase-9 (z-LEHD-FMK) could reverse those effect of TSN (P<0.05) and also reverse the survival rates of CAVO-3 and A2870 cells (P<0.05). In addition, TSN could increase the expression of Bcl-2, Bax and Cyto-C protein in A2870 cells notably, which could be reversed mostly by Caspase-9 inhibitor (z-LEHD-FMK)(P<0.05). CONCLUSION: TSN could induce the apoptosis in human ovarian cancer cell through up-regulating the activities of Caspase-3 and Caspase-9, and then activates the mitochondrial pathway.
ABSTRACT
AIM:To investigate the effect of toosendanin(TSN)on invasion and migration abilities of human ovarian cancer cells and the related mechanism.METHODS:The human ovarian cancer cell lines CAVO-3 and SKVO-3 were treated with TSN at different concentrations.The cell viabilty at 12,24,48,72 and 96 h after TSN treatment was measured by CCK-8 assay.Scratch wound healing assay and Transwell assay were employed to measure the invasion and migration abilities of CAVO-3 cells.The protein expression of nuclear factor-κB(NF-κB)p65,E-cadherin,N-cadherin,vimentin and Snail was determined by Western blot.RESULTS:TSN significantly inhibited the viability of CAVO-3 and SKVO-3 cells(P<0.05 ).Compared with control group ,the migration and invasion abilities of CAVO-3 cells in TSN group decreased significantly(P<0.05).In addition,the expression of NF-κB p65 and E-cadherin protein increased no-tably,followed with N-cadherin,vimentin and Snail protein decreased significantly(P<0.05).However,the inhibitor of NF-κB BAY11-7082 reversed the impact above.Compared with TSN group ,the migration and invasion abilities in TSN +BAY11-7082 group increased significantly(P<0.05).The protein expression of E-cadherin also decreased notably ,fol-lowed with the protein expression of N-cadherin,vimentin and Snail increased significantly(P<0.05).CONCLUSION:TSN inhibits the invasion and migration abilities of human ovarian cancer cells ,which is related to the inhibition of epitheli-al-mesenchymal transition process mediated by NF-κB/Snail signaling pathway.
ABSTRACT
Many studies have been examined the association of platelet glycoprotein (GP) Ia C807T polymorphism with ischemic stroke (IS) susceptibility. However, the results of these studies are inconsistent. To further assess the effects of GP Ia C807T polymorphism on the risk of IS, a meta-analysis was performed in a separate ethnic group. Relevant studies were identified using PubMed and Chinese databases through January 2017. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the strength of the associations. Finally, 13 studies contained 2438 IS cases and 2308 controls included. In the total analyses, a significantly elevated risk of IS was associated with all variants of GP Ia C807T in the Chinese population (T vs C: OR = 1.24, 95% CI = 1.09-1.40; TT vs CC: OR = 1.59, 95% CI = 1.17-2.15; TT and CT combined vs CC: OR = 1.32, 95% CI = 1.09-1.59; TT vs CC and CT: OR = 1.35, 95% CI = 1.04-1.76). In the subgroup analyses stratified by ethnicity and geographic areas, it revealed the significant results in Chinese Han and in South China. This meta-analysis provides the evidence that GP Ia C807T polymorphism may contribute to the IS development in the Chinese population, especially in South China, and further studies in other ethic groups are required for definite conclusions.
Subject(s)
Integrin alpha2/genetics , Polymorphism, Genetic/genetics , Stroke/genetics , Genetic Predisposition to Disease/genetics , Genotype , Humans , Odds RatioABSTRACT
This case-control study was designed to establish a new risk-prediction model for primary stroke using Framingham stroke profile (FSP), cerebral vascular hemodynamic indexes (CVHI) and plasma inflammatory cytokines including hs-CRP, IL-6, TNF-α and Lp-PLA2. A total of 101 primary stroke patients admitted to Dongguan Houjie Hospital between August 2014 and June 2015 were assigned into the case group, and 156 age- and gender-matched healthy subjects from the Houjie Community were allocated into the control group. The prognostic values of FSP, CVHI and inflammatory cytokines including high sensitive C-reactive protein (hs-CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and lipoprotein-associated phospholipase A2 (Lp-PLA2) were assessed by multivariate logistic regression analysis. Seven risk-prediction models (FSP, CVHI, inflammatory cytokine, FSP+CVHI, FSP+inflammatory cytokine, CVHI+inflammatory cytokine, CVHI+FSP+inflammatory cytokine) were successfully established and the prognostic values were statistically compared by ROC curve and Z test. For FSP, the stroke risk was significantly elevated by 2.85 times when the FSP score was increased by 1 level (P=0.043), increased by 3.25 times for CVHI (P=0.036), 6.53 times for IL-6 (P=0.003), and 7.75 times for Lp-PLA2 (P=0.000). The sensitivity of FSP+CVHI+inflammatory cytokine and CVHI+inflammatory cytokine models was higher than 90%. For model specificity, the specificity of FSP+CVHI+inflammatory cytokine model alone exceeded 90%. FSP, CVHI, IL-6 and Lp-PLA2 are independent risk factors of stroke. Integrating IL-6 and Lp-PLA2 into the models can significantly enhance the risk prediction accuracy of primary stroke. Combined application of FSP+CVHI+inflammatory cytokine is of potential for risk prediction of primary stroke.
Subject(s)
Cerebrovascular Circulation/physiology , Cytokines/blood , Cytokines/metabolism , Stroke/epidemiology , Stroke/metabolism , 1-Alkyl-2-acetylglycerophosphocholine Esterase/blood , Aged , Aged, 80 and over , C-Reactive Protein/metabolism , Case-Control Studies , China/epidemiology , Female , Hemodynamics , Humans , Incidence , Interleukin-6/blood , Male , Middle Aged , Models, Biological , Prognosis , Risk Factors , Sensitivity and Specificity , Severity of Illness Index , Tumor Necrosis Factor-alpha/bloodABSTRACT
BACKGROUND/AIMS: This study aimed to investigate whether Salvianolic acid A (Sal A) conferred cardiac protection against Arsenic trioxide (ATO)-induced cardiotoxicity in H9c2 cells by inhibiting MAPK pathways activation. METHODS: H9c2 cardiac cells were exposed to 10 µM ATO for 24 h to induce cytotoxicity. The cells were pretreated with Sal A for 4 h before exposure to ATO. Cell viability was determined utilizing the MTT assay. The percentage of apoptosis was measured by a FITC-Annexin V/PI apoptosis kit for flow cytometry. Mitochondrial membrane potential (∆Ψm) was detected by JC-1. The intracellular ROS levels were measured using an Image-iTTM LIVE Green Reactive Oxygen Species Detection Kit. The apoptosis-related proteins and the MAPK signaling pathways proteins expression were quantified by Western blotting. RESULTS: Sal A pretreatment increased cell viability, suppressed ATO-induced mitochondrial membrane depolarization, and significantly altered the apoptotic rate by enhancing endogenous antioxidative enzyme activity and ROS generation. Signal transduction studies indicated that Sal A suppressed the ATO-induced activation of the MAPK pathway. More importantly, JNK, ERK, and p38 inhibitors mimicked the cytoprotective activity of Sal A against ATO-induced injury in H9c2 cells by increasing cell viability, up-regulating Bcl-2 protein expression, and down-regulating both Bax and caspase-3 protein expression. CONCLUSION: Sal A decreases the ATO-induced apoptosis and necrosis of H9c2 cells, and the underlying mechanisms of this protective effect of Sal A may be connected with the MAPK pathways.
Subject(s)
Cardiotonic Agents/pharmacology , MAP Kinase Signaling System/drug effects , Oxides/toxicity , Animals , Apoptosis/drug effects , Arsenic Trioxide , Arsenicals , Caffeic Acids , Cell Line , Cell Survival/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , Lactates , Necrosis , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Rats , Up-Regulation/drug effectsABSTRACT
BACKGROUND: We have previously shown that Elatoside C reduces cardiomyocyte apoptosis during ischaemia/reperfusion (I/R). Here, we investigated whether Elatoside C improves heart function in isolated rat hearts subjected to I/R and elucidated the potential mechanisms involved in Elatoside C-induced protection. METHODS AND RESULTS: Isolated rat hearts were subjected to global ischaemia followed by reperfusion in the absence or presence of Elatoside C. We found that Elatoside C significantly attenuated cardiac dysfunction and depressed oxidative stress induced by I/R. Consistently, Elatoside C prevented I/R-induced mitochondrial dysfunction, which was evident by the inhibition of mitochondrial ROS production, mitochondrial permeability transition pore (mPTP) opening, cytochrome c release from the mitochondria and Bax translocation. Moreover, Elatoside C improved abnormal calcium handling during I/R, including increasing sarcoplasmic reticulum Ca(2+) ATPase (SERCA2) activity, alleviating [Ca(2+)]ER depletion, and reducing the expression levels of ER stress protein markers. All of these protective effects of Elatoside C were partially abolished by the PI3K/Akt inhibitor LY294002, ERK1/2 inhibitor PD98059, and JAK2/STAT3 inhibitor AG490. Further assessment in isolated cardiomyocytes showed that Elatoside C maintained the Ca(2+) transients and cell shortening against I/R. CONCLUSIONS: Elatoside C protects against cardiac injury during I/R by attenuating oxidative stress and [Ca(2+)]i overload through the activation of both the reperfusion injury salvage kinase (RISK) pathway (including PI3K/Akt and ERK1/2) and the survivor activating factor enhancement (SAFE) pathway (including JAK2/STAT3) and, subsequently, inhibiting the opening of mPTPs.
Subject(s)
Calcium/metabolism , Homeostasis/drug effects , Intracellular Fluid/metabolism , Myocardial Reperfusion Injury/prevention & control , Myocytes, Cardiac/metabolism , Oxidative Stress/drug effects , Saponins/pharmacology , Animals , Apoptosis/drug effects , Blotting, Western , Disease Models, Animal , Male , Myocardial Reperfusion Injury/metabolism , Myocardial Reperfusion Injury/pathology , Myocytes, Cardiac/pathology , Rats , Rats, Sprague-DawleyABSTRACT
Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases. MMPs can degrade and remodel extracellular matrix, also active or inactive many molecules attaching to matrix including receptors, growth factors and cytokines, so that injury-induced MMPs can change the extracellular environment to affect the axonal regeneration in central nervous system. In this review, with spinal cord injury (SCI) as an example we discuss the effects of MMPs on inflammation, neuronal viability, extracellular molecules, glial scar and axonal remyelination, which are all important to axonal regeneration.
Subject(s)
Axons , Cicatrix , Extracellular Matrix , Matrix Metalloproteinases , Nerve Regeneration , Neuroglia , Spinal Cord InjuriesABSTRACT
Endoplasmic reticulum (ER) stress-induced apoptosis has been suggested to contribute to myocardial ischemia-reperfusion (I/R) injury. Elatoside C is one of the major triterpenoid compounds isolated from Aralia elata that is known to be cardioprotective. However, its effects on I/R injury to cardiac myocytes have not been clarified. This study aimed to investigate the possible protective effect of Elatoside C against hypoxia/reoxygenation (H/R)-induced H9c2 cardiomyocyte injury and its underlying mechanisms. H9c2 cardiomyocytes were subjected to H/R in the presence of Elatoside C. Our results showed that Elatoside C (25 µM) treatment provided significant protection against H/R-induced cell death, as evidenced by improved cell viability, maintained mitochondrial membrane potential, diminished mitochondrial ROS, and reduced apoptotic cardiomyocytes (P < 0.05). These changes were associated with the inhibition of ER stress-associated apoptosis markers (GRP78, CHOP, Caspase-12 and JNK), as well as the increased phosphorylation of STAT3 and an increased Bcl2/Bax ratio. Moreover, these effects of Elatoside C were prevented by the STAT3 inhibitor Stattic. Taken together, these results suggested that Elatoside C can alleviate H/R-induced cardiomyocyte apoptosis most likely by activating the STAT3 pathways and reducing ER stress-associated apoptosis.
Subject(s)
Apoptosis/drug effects , Cardiotonic Agents/pharmacology , Endoplasmic Reticulum Stress/drug effects , Myocytes, Cardiac/drug effects , STAT3 Transcription Factor/metabolism , Saponins/pharmacology , Triterpenes/pharmacology , Animals , Aralia , Cell Hypoxia , Cell Line , Myocardial Reperfusion Injury/metabolism , Myocardial Reperfusion Injury/pathology , Myocytes, Cardiac/metabolism , Myocytes, Cardiac/pathology , RatsABSTRACT
<p><b>OBJECTIVE</b>To study the effect of Jiedu Huayu Recipe containing serum on the expressions of survivin gene of leukemia K562/A02 cell apoptosis and the expression of nuclear factor kappa B (NF-kappaB).</p><p><b>METHODS</b>K562/A02 cells were divided into six groups. An equal volume of calf serum, rabbit serum, high, middle, and low dose Jiedu Huayu Recipe containing serum, and interferon was respectively added. K562 sensitive cell strain was set up as the control. The expressions of survivin in K562/A02 cells after treated with Jiedu Huayu Recipe containing serum were detected using semi-quantitative reverse transcriptase polymerase chain reaction method. The expressions of NF-kappaB/P65, NF-kappaB/P50, and IKBa were detected using Westem blot method.</p><p><b>RESULTS</b>The survivin resistant gene was highly expressed in K562/A02 cells. After treated by Jiedu Huayu Recipe containing serum, the expressions of survivin could be obviously lowered in the high and middle dose Jiedu Huayu Recipe containing serum groups as well as the interferon control group. NF-kappaB was also highly expressed in K562/A02 resistant cells. After treated by Jiedu Huayu Recipe containing serum, the expressions of P65, P50, and IkappaBalpha decreased to some degrees. The decrement was the most obvious in the middle and high dose Jiedu Huayu Recipe containing serum groups. The decreased expressions of P65 and P50 in the high dose Jiedu Huayu Recipe containing serum group were higher than that in the interferon control group (P<0.05). The decrement of IkappaBalpha was equivalent to that of the interferon control group (P>0.05).</p><p><b>CONCLUSION</b>Jiedu Huayu Recipe could block the NF-kappaB signal pathway of K562/A02 cells and reverse drug resistance by influencing the expression of NF-kappaB-survivin genes.</p>
Subject(s)
Humans , Apoptosis , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Drugs, Chinese Herbal , Pharmacology , Inhibitor of Apoptosis Proteins , Metabolism , K562 Cells , NF-kappa B , Metabolism , SerumABSTRACT
The study was purposed to investigate the apoptosis of HL-60 cells induced by recombinant common buckwheat trypsin inhibitor (rBTI) and its mechanism. The inhibition rate of rBTI on HL-60 cells was detected by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide); the morphology of HL-60 nuclei was observed by fluorescence microscopy; the apoptosis cells of HL-60 detected by agarose gel electrophoresis and the changes of apoptosis rate was assayed by flow cytometry (FCM), when the HL-60 cells were treated with different concentration of rBTI for 24 hours. The results showed that the growth of HL-60 cells was inhibited evidently after treatment with rBTI in a dose-dependent manner, but there were minimal effects on normal human peripheral blood mononuclear cells (PBMNCs). The nuclei of HL-60 cells showed the characteristics of apoptosis, the analysis by flow cytometry indicated that the apoptosis rate of HL-60 cells was 52% after treatment with rBTI (100 microg/ml), DNA analyzed by agarose gel electrophoresis showed "ladder" pattern. It is concluded that rBTI obviously inhibits growth of HL-60 and induces its apoptosis which provides a foundation for use of recombinant common buckwheat trypsin inhibitor to cure the acute myeloid leukemia.