ABSTRACT
Severe lumbosacral pain, paraparesis or paraplegia, and urinary incontinence are common but frustrating problems in dogs with lumbosacral spinal cord injury (SCI). The surgical interventions including stabilization and decompression may not restore satisfying neurological functions in severe SCI. Adipose tissue-derived mesenchymal stem cells (Ad-MSCs) show benefits in immunomodulation, anti-inflammation, and promotion of axonal growth and remyelination, and also display efficacy in several diseases in veterinary medicine. In this report, four dogs presented with fracture of sacrum vertebrae or fracture of seventh lumbar and lumbosacral displacement after road traffic accidents. The clinical signs include lumbosacral pain (4/4), paraparesis (3/4), paraplegia (1/4), and urinary incontinence (4/4). All dogs were treated by surgical decompression with or without stabilization 1 to 7 weeks after trauma. Allogeneic canine Ad-MSCs (cAd-MSCs) were injected locally on nerve roots through the surgical region in all dogs. One dose of intravenous transplantation and 4 doses of local transplantation were also performed within 8 weeks after the surgery separately. All dogs showed significant neurological improvements with normal ambulatory ability (4/4) and urinary control (3/4) 3 months after the surgery and the first cAd-MSCs transplantation. No side effect was related to multiple cAd-MSCs transplantations during 6 months monitoring in all dogs. In conclusion, multiple cAd-MSCs transplantations could be a recommended treatment combined with surgery in dogs with lumbosacral SCI.
Subject(s)
Hematopoietic Stem Cell Transplantation , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Spinal Cord Injuries , Animals , Dogs , Spinal Cord/surgery , Spinal Cord Injuries/surgery , Spinal Cord Injuries/veterinaryABSTRACT
Polyalthia belong to the Annonaceae family and are a type of evergreen tree distributed across many tropical and subtropical regions. Polyalthia species have been used long term as indigenous medicine to treat certain diseases, including fever, diabetes, infection, digestive disease, etc. Recent studies have demonstrated that not only crude extracts but also the isolated pure compounds exhibit various pharmacological activities, such as anti-oxidant, anti-microbial, anti-tumor, anti-cancer, etc. It is known that the initiation of cancer usually takes several years and is related to unhealthy lifestyle, as well as dietary and environmental factors, such as stress, toxins and smoking. In fact, natural or synthetic substances have been used as cancer chemoprevention to delay, impede, or even stop cancer growing. This review is an attempt to collect current available phytochemicals from Polyalthia species, which exhibit anti-cancer potentials for chemoprevention purposes, providing directions for further research on the interesting agents and possible clinical applications.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Antioxidants/pharmacology , Phytochemicals/pharmacology , Polyalthia/chemistry , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/isolation & purification , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Antioxidants/chemistry , Antioxidants/isolation & purification , Humans , Molecular Structure , Phytochemicals/chemistry , Phytochemicals/isolation & purificationABSTRACT
Inflammatory bowel disease (IBD) is general term for ulcerative colitis and Crohn's disease, which is chronic intestinal and colorectal inflammation caused by microbial infiltration or immunocyte attack. IBD is not curable, and is highly susceptible to develop into colorectal cancer. Finding agents to alleviate these symptoms, as well as any progression of IBD, is a critical effort. This study evaluates the anti-inflammation and anti-tumor activity of 16-hydroxycleroda-3,13-dien-15,16-olide (HCD) in in vivo and in vitro assays. The result of an IBD mouse model induced using intraperitoneal chemical azoxymethane (AOM)/dextran sodium sulfate (DSS) injection showed that intraperitoneal HCD adminstration could ameliorate the inflammatory symptoms of IBD mice. In the in vitro assay, cytotoxic characteristics and retained signaling pathways of HCD treatment were analyzed by MTT assay, cell cycle analysis, and Western blotting. From cell viability determination, the IC50 of HCD in Caco-2 was significantly lower in 2.30 µM at 48 h when compared to 5-fluorouracil (5-FU) (66.79 µM). By cell cycle and Western blotting analysis, the cell death characteristics of HCD treatment in Caco-2 exhibited the involvement of extrinsic and intrinsic pathways in cell death, for which intrinsic apoptosis was predominantly activated via the reduction in growth factor signaling. These potential treatments against colon cancer demonstrate that HCD could provide a promising adjuvant as an alternative medicine in combating colorectal cancer and IBD.
Subject(s)
Apoptosis , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/pathology , Diterpenes, Clerodane/therapeutic use , Inflammatory Bowel Diseases/drug therapy , Animals , Apoptosis/drug effects , Azoxymethane , Biomarkers, Tumor/metabolism , Caco-2 Cells , Cell Cycle/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Dextran Sulfate , Diterpenes, Clerodane/pharmacology , Fluorouracil/pharmacology , Fluorouracil/therapeutic use , HT29 Cells , Humans , Inflammation/drug therapy , Inflammation/pathology , Intestines/pathology , Male , Mice, Inbred C57BLABSTRACT
BACKGROUND: Renal cell carcinoma (RCC) is well known that it cannot be treated with traditional chemotherapy or radiotherapy. 16-Hydroxycleroda-3,13-dien-15,16-olide (CD), isolated from Polyalthia longifolia Benth. & Hook. f. var. pendula had been reported to display significant efficacy against cancer cell lines. PURPOSE: To determine the anti-tumour activities of CD in two clear cell type RCC (ccRCC) cell lines (A-498 and 786-O). In addition, the underlying mechanisms were also examined. METHODS: The cell viabilities of CD-treated ccRCC cells were examined by MTT assay. The apoptotic features were confirmed by acridine orange and ethidium bromide staining. 2',7'-dichlorofluorescin diacetate was used to check reactive oxygen species (ROS) involvement. Mitochondria membrane potential (MMP) were determined by using fluorescent dyes, rhodamine 123 and 5',6,6'-tetrachloro-1,1',3,3'-tetraethyl benzimidazolylcarbocyanine iodide (JC-1). Proapoptotic, anti-apoptotic proteins and intracellular signaling molecules involved in CD-induced apoptosis were examined by Western blot analysis. RESULTS: CD inhibited both 786-O and A-498 cell proliferation and induced a series apoptotic characteristics expressions, ROS accumulation, caspase-3 activation as well as poly-(ADP-ribose) polymerase cleavage in both ccRCC cells. Additionally, CD caused MMP reduction and cytochrome c release from mitochondria as well as inhibition of anti-apoptotic proteins, including B cell lymphoma 2 and heat shock protein 70. Mechanically, we address that CD suppressed cell proliferation and induced apoptosis via induction of FOXO3a as well as decreased phosphorylation of Akt, mTOR, MEK/ERK and their downstream molecules, cMyc and hypoxia inducible factor 2α expression in a concentration- and time-dependent trend. CONCLUSION: CD caused cell death through ROS overproduction and induction of mitochondria-dependent apoptotic pathway in ccRCC cells that accompanied with multiple oncogenic signals inactivation.
Subject(s)
Apoptosis/drug effects , Diterpenes/pharmacology , Kidney Neoplasms/drug therapy , MAP Kinase Signaling System/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Antineoplastic Agents, Phytogenic/pharmacology , Caspase 3/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cytochromes c/metabolism , Humans , Kidney Neoplasms/metabolism , Kidney Neoplasms/pathology , Membrane Potential, Mitochondrial/drug effects , Mitochondria/drug effects , Mitochondria/metabolism , Reactive Oxygen Species/metabolism , TOR Serine-Threonine Kinases/metabolismABSTRACT
Migration of vascular smooth muscle cells (VSMCs) contributes to intimal hyperplasia and other vascular diseases. Caveolin-1 (Cav-1) has been recognized as a proliferative inhibitor of VSMCs and is likely to be an important regulator of VSMC migration. The underlying mechanism of pyrogallol on the VSMC migration is not fully understood. This study attempted to dissect the role of Cav-1 and matrix metalloproteinase (MMP) in VSMC migration and to investigate the effect of pyrogallol on VSMC mobility during carotid artery ligation mice. The mRNA expression of MMP-3 and MMP-13 was down-regulated in cultured VSMC prepared from Cav-1-deficient (Cav-1 KO) mice whereas MMP-14 expression was up-regulated. Pyrogallol effectively inhibited the migration of Cav-1 KO VSMC by promoting the expression of tissue inhibitors of metalloproteinase (TIMP)-2. Pyrogallol also inhibited the migration of Cav-1 wild type (WT) VSMC, however, by increasing TIMP-1 expression and repressing MMP-2 activity. In a parallel in vivo study, intra-peritoneal (ip) of pyrogallol to carotid artery ligated mice significantly suppressed intima formation in mice carotid artery. Furthermore, the proMMP-9 activity in pyrogallol-treated mice serum significantly increased from Day 0 to Day 2 and decreased from Day 2 to Day 7 in a time-dependent manner. In addition, WT mice treated with pyrogallol had significantly reduced neointima formation, whereas no differences were observed in Cav-1 knock out (KO) mice. These results suggest that pyrogallol not only inhibited VSMC migration but also effectively diminishes the severity of neointima hyperplasia, implying that pyrogallol possesses potential anti-atherogenic effects for the treatment of vascular diseases.
Subject(s)
Carotid Arteries/drug effects , Caveolin 1/metabolism , Cell Movement/drug effects , Matrix Metalloproteinases/metabolism , Myocytes, Smooth Muscle/drug effects , Neointima/prevention & control , Pyrogallol/therapeutic use , Animals , Carotid Arteries/pathology , Caveolin 1/genetics , Cell Culture Techniques , Cell Survival/drug effects , Cells, Cultured , Disease Models, Animal , Male , Matrix Metalloproteinases/genetics , Meliaceae/chemistry , Mice, Inbred C57BL , Mice, Knockout , Myocytes, Smooth Muscle/metabolism , Myocytes, Smooth Muscle/pathology , Neointima/etiology , Neointima/metabolism , Neointima/pathology , Pyrogallol/isolation & purification , Pyrogallol/pharmacology , Tunica Intima/drug effects , Tunica Intima/pathologyABSTRACT
Toona sinensis (TS) is a type of deciduous tree, which is distributed widely in Asia and used as a traditional herb medicine. Previously, we demonstrated that aqueous extracts of TS leaves (TSL-1) induce apoptosis in two clear types of human renal carcinoma cells (ccRCC) via mitochondria-dependent pathway. In this study, we further investigated the more detailed mechanism of TSL-1-induced antitumor effects on ccRCCs. TSL-1 treatment arrested ccRCC cells in G0/G1 phase through the decrease of cyclin D1, cyclin-dependent kinase (CDK)2, and CDK4 as well as induction of p53 and FOXO3a protein expressions. On the other hand, the inhibitory effects of TSL-1 on migration were also observed in 786-O and A-498 cells. Mechanically, we presented that TSL-1 could suppress cell cycle progression and motility via inhibiting the phosphorylation of JAK2/stat3, Akt, MEK/ERK, and mTOR in a concentration- and time-dependent manner. Moreover, we found that TSL-1 inhibited p21, HIF-2α, c-Myc, VEGF, and MMP9 protein expressions in both cell lines. In conclusion, these findings suggested that TS-induced apoptosis and its antimigration activity in ccRCC cells were accompanied by inactivation of several oncogenic pathways.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Kidney Neoplasms/drug therapy , Meliaceae/chemistry , Plant Extracts/pharmacology , Antineoplastic Agents, Phytogenic/chemistry , Apoptosis/drug effects , Basic Helix-Loop-Helix Transcription Factors/metabolism , Cell Line, Tumor , Cell Movement/drug effects , Cyclins/metabolism , Humans , Janus Kinase 2/metabolism , Kidney Neoplasms/metabolism , Kidney Neoplasms/pathology , MAP Kinase Kinase Kinases/metabolism , Plant Extracts/chemistry , Plant Leaves/chemistry , Proto-Oncogene Proteins c-akt/metabolism , STAT3 Transcription Factor/metabolism , Signal Transduction/drug effects , TOR Serine-Threonine Kinases/metabolismABSTRACT
The alpha-glucosidase inhibitor is a common oral anti-diabetic drug used for controlling carbohydrates normally converted into simple sugars and absorbed by the intestines. However, some adverse clinical effects have been observed. The present study seeks an alternative drug that can regulate the hyperglycemia by down-regulating alpha-glucosidase and alpha-amylase activity by molecular docking approach to screen the hyperglycemia antagonist against alpha-glucosidase and alpha-amylase activities from the 47 natural compounds. The docking data showed that Curcumin, 16-hydroxy-cleroda-3,13-dine-16,15-olide (16-H), Docosanol, Tetracosanol, Antroquinonol, Berberine, Catechin, Quercetin, Actinodaphnine, and Rutin from 47 natural compounds had binding ability towards alpha-amylase and alpha-glucosidase as well. Curcumin had a better biding ability of alpha-amylase than the other natural compounds. Analyzed alpha-glucosidase activity reveals natural compound inhibitors (below 0.5 mM) are Curcumin, Actinodaphnine, 16-H, Quercetin, Berberine, and Catechin when compared to the commercial drug Acarbose (3 mM). A natural compound with alpha-amylase inhibitors (below 0.5 mM) includes Curcumin, Berberine, Docosanol, 16-H, Actinodaphnine/Tetracosanol, Catechin, and Quercetin when compared to Acarbose (1 mM). When taken together, the implication is that molecular docking is a fast and effective way to screen alpha-glucosidase and alpha-amylase inhibitors as lead compounds of natural sources isolated from medicinal plants.
Subject(s)
Diterpenes/chemistry , Glycoside Hydrolase Inhibitors/chemistry , Hypoglycemic Agents/chemistry , alpha-Amylases/antagonists & inhibitors , alpha-Glucosidases/chemistry , Acarbose/chemistry , Berberine/chemistry , Biological Products/chemistry , Curcumin/chemistry , High-Throughput Screening Assays , Humans , Molecular Docking Simulation , Quercetin/chemistry , Recombinant Proteins/chemistry , User-Computer Interface , alpha-Amylases/chemistryABSTRACT
Autophagy, a lysosomal pathway to maintain cellular homeostasis, is mediated via the mammalian target of rapamycin (mTOR)-dependent pathways. Hepatic stellate cells (HSCs), previously termed fat- or vitamin A-storing cells, can transdifferentiate into myofibroblast-like cells and are the most relevant cell type for overproduction of extracellular matrix (ECM) and development of liver fibrosis during injury. However, the role of autophagy in fat metabolism of HSCs remains unclear. This study investigates the regulatory effect of natural compounds on fatty acid-induced autophagy pathways of nonchemical-induced HSC (NHSC) and thioacetamide-induced HSC. Oleic acid (OA) and palmitic acid (PA) have shown a significant effect on cell proliferation with oil red O staining and Western blot confirming that OA and PA induce fat storage ability and autophagy protein expression in NHSC. Natural compounds rutin, curcumin, antroquinonol and benzyl cinnamate treatment have shown no effect on the autophagy protein expression. Nevertheless, cells pretreated with OA and PA then treated with rutin, curcumin, antroquinonol and benzyl cinnamate could significantly induce the light chain I/II (LC3 I/II) protein expression. In mTOR-dependent pathway, the PI3K-Class I, Akt, and p-mTOR proteins were decreased with PA treatment. However, there were no significant changes in PI3K-Class III and Beclin-1 protein expressions found to imply that this autophagy is unrelated to the mTOR-independent pathway. Taken together, the present study unveils rutin and curcumin as a possible effective stimulation for fatty acid-induced autophagy via mTOR-dependent pathways in NHSC. We further suggest the benefits of these natural compounds for alleviating liver fibrosis.
Subject(s)
Autophagy/drug effects , Biological Products/pharmacology , Fatty Acids/pharmacology , Hepatic Stellate Cells/drug effects , Animals , Cells, Cultured , Drug Synergism , Hepatic Stellate Cells/immunology , Hepatic Stellate Cells/metabolism , Lipids/biosynthesis , Male , Rats , Rats, Sprague-DawleyABSTRACT
Toona sinensis is one of the most popular vegetarian cuisines in Taiwan and it has been shown to possess antioxidant, antiangiogenic, and anticancer properties. In this study, we investigated the antiatherosclerotic potential of aqueous leaf extracts from Toona sinensis (TS; 25-100 µg/mL) and its major bioactive compound, gallic acid (GA; 5 µg/mL), in LPS-treated rat aortic smooth muscle (A7r5) cells. We found that pretreatment with noncytotoxic concentrations of TS and GA significantly inhibited inflammatory NO and PGE2 production by downregulating their precursors, iNOS and COX-2, respectively, in LPS-treated A7r5 cells. Furthermore, TS and GA inhibited LPS-induced intracellular ROS and their corresponding mediator, p47(phox). Notably, TS and GA pretreatment significantly inhibited LPS-induced migration in transwell assays. Gelatin zymography and western blotting demonstrated that treatment with TS and GA suppressed the activity or expression of MMP-9, MMP-2, and t-PA. Additionally, TS and GA significantly inhibited LPS-induced VEGF, PDGF, and VCAM-1 expression. Further investigation revealed that the inhibition of iNOS/COX-2, MMPs, growth factors, and adhesion molecules was associated with the suppression of NF-κB activation and MAPK (ERK1/2, JNK1/2, and p38) phosphorylation. Thus, Toona sinensis may be useful for the prevention of atherosclerosis.
Subject(s)
Cell Movement/drug effects , Inflammation/drug therapy , Meliaceae/chemistry , Muscle, Smooth, Vascular/pathology , Myocytes, Smooth Muscle/pathology , NF-kappa B/metabolism , Plant Extracts/pharmacology , Reactive Oxygen Species/metabolism , Animals , Cell Line , Cell Survival/drug effects , Cyclooxygenase 2/metabolism , Dinoprostone/biosynthesis , Down-Regulation/drug effects , Eye Proteins/metabolism , Gallic Acid/pharmacology , I-kappa B Proteins/metabolism , Lipopolysaccharides , MAP Kinase Signaling System/drug effects , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/enzymology , NADPH Oxidases/metabolism , NF-KappaB Inhibitor alpha , Nerve Growth Factors/metabolism , Nitric Oxide/biosynthesis , Nitric Oxide Synthase Type II/metabolism , Plant Extracts/therapeutic use , Rats , Serpins/metabolism , Signal Transduction/drug effects , Tissue Plasminogen Activator/metabolism , Vascular Endothelial Growth Factor A/metabolismABSTRACT
SCOPE: We investigated whether rutin, a flavonoid isolated from Toona sinensis Roem, has the ability to enhance insulin-dependent receptor kinase (IRK) activity and glucose transporter 4 (GLUT4) translocation in differentiated myotubes. We also tested the effects of rutin treatment in insulin-resistant mice using an oral glucose tolerance test (OGTT). METHODS AND RESULTS: Rutin potentiated insulin receptor kinase (IRK) phosphorylation when IRK autophosphorylation was triggered by insulin in differentiated myotubes. Co-treatment of cells with rutin and insulin attenuated S961-mediated inhibition of insulin-dependent GLUT4 translocation. In S961-treated C57BL/6 mice, an in vivo model of insulin resistance and type 2 diabetes, rutin treatment showed a normoglycemic effect in the OGTT. CONCLUSION: This study shows evidence that rutin may serve as a potential agent for glycemic control through enhancement of IRK activity, thereby inducing the insulin signaling pathway causing increased GLUT4 translocation and increased glucose uptake.
Subject(s)
Glucose Transporter Type 4/metabolism , Plant Extracts/pharmacology , Receptor, Insulin/metabolism , Rutin/pharmacology , Animals , Blood Glucose , Cells, Cultured , Diabetes Mellitus, Type 2/drug therapy , Glucose Tolerance Test , Insulin/metabolism , Insulin Resistance , Male , Meliaceae/chemistry , Mice , Mice, Inbred C57BL , Muscle Fibers, Skeletal/cytology , Muscle Fibers, Skeletal/drug effects , Myoblasts, Skeletal/cytology , Myoblasts, Skeletal/drug effects , Phosphorylation , Signal TransductionABSTRACT
SCOPE: Diabetes is a critical factor for atherosclerosis, as hyperglycemia induces vascular smooth muscle cell (VSMC) proliferation and migration and subsequently contributes to the formation of atherosclerotic lesions. This study investigates whether resveratrol plays a regulatory role in the proliferation and migration of VSMCs under high glucose induction to imitate a hyperglycemic condition. METHODS AND RESULTS: Resveratrol inhibited the migration of VSMCs in the wound-healing assay and the formation of lamellipodia and filopodia as assessed by atomic force microscopy scanning. Resveratrol suppressed the mRNA expression of c-Src, Rac1, cdc42, IRS-1, MEKK1, MEKK4, and mitogen-activated protein kinase along with the protein levels of c-Src, p-Src, and cdc42 in VSMCs. Resveratrol decreased the level of p-FAK protein under normal glucose conditions. Resveratrol could inhibit the activities of matrix metalloproteinase (MMP) 2 and MMP 9 as shown by zymography. Moreover, resveratrol also regulated the mitogen-activated protein kinase pathway and MMP activities of VSMC migration under the high glucose condition. CONCLUSION: The antimigratory effects of resveratrol by reduced MMP expression through the inhibition of Rac1, p-FAK, and lamellipodia formation and the activation of p-AKT and p-ERK1/2 suggest that resveratrol is a potential compound for the treatment of vascular diseases via the regulation of VSMC migration.
Subject(s)
Focal Adhesion Protein-Tyrosine Kinases/metabolism , Glucose/adverse effects , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/drug effects , Stilbenes/pharmacology , Animals , CSK Tyrosine-Protein Kinase , Cell Movement/drug effects , Cell Survival/drug effects , Cells, Cultured , MAP Kinase Signaling System/drug effects , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Mice , Mice, Inbred C57BL , Microscopy, Atomic Force , Mitogen-Activated Protein Kinases/genetics , Mitogen-Activated Protein Kinases/metabolism , Muscle, Smooth, Vascular/drug effects , Pseudopodia/drug effects , Pseudopodia/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Resveratrol , Wound Healing/drug effects , src-Family Kinases/genetics , src-Family Kinases/metabolismABSTRACT
BACKGROUND: Focal adhesion kinase (FAK) is a nonreceptor protein tyrosine plays an important role in a number of cell signaling pathways, including cell migration, proliferation, and cell survival. This study was aimed to identify novel and specific inhibitors from natural compounds via molecular docking of FAK (Y397). METHODS: The 3D structure of FAK (PDB ID: 2AL6) was used for docking 109 natural compounds. Based on high affinity and energy interaction, four of ten candidate compounds, 16-hydroxy-cleroda-3,13-dien-16,15-olide (HCD), curcumin, quercetin, and catechin hydrate, were hit, and the inhibitory activity against FAK was validated in these compounds in C6 glioma and N18 neuroblastoma cell lines. RESULTS: HCD showed a potential effect on cell viability by MTT assay and cell arrest in the G0-G1 phase, and a TUNEL assay confirmed further apoptosis. Treatment with HCD decreased anti-apoptotic proteins and increased pro-apoptotic proteins. Atomic force microscopy data depicted that the formation of filopodia on the intracellular surface decreased in treated cells compared with the control. Zymography showed that HCD inhibited the activity of MMP-2 and MMP-9. The protein levels of FAK, pFAK, Rac1 and Cdc42, which are the key regulators for the formation of filopodia, were decreased. Additionally, HCD regulated the expression of epithelial mesenchymal transition proteins. CONCLUSIONS: HCD effectively interacted at the autophosphorylation site of FAK and interaction analysis indicated an H-bond with the Arg 86 and Arg 125 residues. GENERAL SIGNIFICANCE: This study suggests that HCD could be a potential inhibitor of FAK and could be used for anti-tumorigenesis and anti-metastasis treatments.
Subject(s)
Antineoplastic Agents/pharmacology , Diterpenes/pharmacology , Focal Adhesion Protein-Tyrosine Kinases/antagonists & inhibitors , Molecular Docking Simulation , Protein Kinase Inhibitors/pharmacology , Animals , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line, Tumor , Mice , Models, Molecular , Phosphorylation , Rats , Signal Transduction/drug effectsABSTRACT
Extract of Toona sinensis (TS) has been reported to have various effects on cultured cell lines, including anti-proliferative activity in cancer cells. We have studied the effects of TS on various human oral squamous carcinoma cell lines (HOSCC), including UM1, UM2, SCC-4, and SCC-9. These cell lines were treated with TS leaf extract and screened for viability, apoptosis, necrosis, and apoptotic gene expression. Normal human oral keratinocytes (NHOK) served as a control for cytotoxic assays. Viability of TS-treated HOSCC was reduced, whereas that of NHOK was not affected. FACScan analysis revealed that the leaf extract induced apoptosis or a combination of apoptosis and necrosis, depending on cell type. Microarray and semi-quantitative RT-PCR analysis for apoptotic-related gene expression revealed that 3,4,5-trihydroxybenzoic acid (gallic acid, one of the major bioactive compounds purified from TS extract) up-regulated pro-apoptotic genes such TNF-α, TP53BP2, and GADD45A, and down-regulated the anti-apoptotic genes Survivin and cIAP1, resulting in cell death. This study suggests that gallic acid, the major bioactive compound present, is responsible for the anti-neoplastic effect of Toona sinensis leaf extract.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Gallic Acid/therapeutic use , Meliaceae/chemistry , Plant Extracts/pharmacology , Plant Leaves/chemistry , Apoptosis/drug effects , Apoptosis/genetics , Apoptosis Regulatory Proteins/metabolism , Carcinoma, Squamous Cell , Cell Cycle Proteins/metabolism , Cell Line, Tumor , Cell Survival/drug effects , Gallic Acid/chemistry , Gene Expression Regulation/drug effects , Humans , Inhibitory Concentration 50 , Keratinocytes/drug effects , Mouth Neoplasms , Nuclear Proteins/metabolism , Oligonucleotide Array Sequence Analysis , Plant Extracts/chemistry , Tumor Necrosis Factor-alpha/metabolismABSTRACT
The stems of Cinnamomum reticulatum Hay (Lauraceae) were extracted with hexane and chloroform successively. A series of new esters, including a mixture of 4-hydroxy-3-methoxyphenethyl derivatives, along with two butanolides, isoobtusilactone A and obtusilactone A, two amides, N-trans-feruloylmethoxytyramine and N-cis-feruloyl-methoxytyramine, three benzenoids, p-hydroxybenzoic acid, syringic acid and vanillic acid, one lignan, (+)-syringaresinol and one steroid, beta-sitostenone, were isolated. The structures of the new esters were elucidated by chemical and physical evidence.
Subject(s)
Cinnamomum/chemistry , Esters/chemistry , Molecular StructureABSTRACT
Prostate cancer, the most frequently diagnosed malignancy in elderly males of the United States, has become a major health issue in Asia. Previous studies have demonstrated that leaf extracts of Toona sinensis Roem. contain cytotoxic activity on several cancer cells including prostate cancer cells. In this study, gallic acid is identified as the major anti-cancer compound in T. sinensis leaf extracts. It is cytotoxic to DU145 prostate cancer cells, through generation of reactive oxygen species (ROS) and mitochondria-mediated apoptosis, which were reversed by antioxidants catalase and N-acetylcysteine. Furthermore, gallic acid is shown to block the growth of DU145 cells at G2/M phases by activating Chk1 and Chk2 and inhibiting Cdc25C and Cdc2 activities. In addition, gallic acid has a synergistic effect with doxorubicin in suppressing the growth of DU145 cells. Taken together, our results suggest that gallic acid has the potential to be developed into an anti-prostate cancer drug and is worthy of further studies.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Gallic Acid/pharmacology , Meliaceae/chemistry , Plant Leaves/chemistry , Reactive Oxygen Species/metabolism , Apoptosis/drug effects , CDC2 Protein Kinase , Cell Cycle/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Checkpoint Kinase 1 , Checkpoint Kinase 2 , Comet Assay , Cyclin B/metabolism , Cyclin-Dependent Kinases , DNA Breaks, Double-Stranded/drug effects , Doxorubicin/pharmacology , Drug Synergism , Flow Cytometry , Gallic Acid/chemistry , Humans , Immunoblotting , Male , Membrane Potential, Mitochondrial/drug effects , Molecular Structure , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/pathology , Protein Kinases/metabolism , Protein Serine-Threonine Kinases/metabolism , cdc25 Phosphatases/metabolismABSTRACT
Pyrogallol, a catechin compound, is an active component of Emblica officinalis extracts and has an anti-proliferative effect on some human cancer cell lines. In our preliminary study, pyrogallol had highly cytotoxic effect on human lung cancer cell lines and less effect on human bronchial epithelium cell line. This study was performed to investigate the beneficial effect of pyrogallol on human lung cancer cell lines - H441 (lung adenocarcinoma) and H520 (lung squamous cell carcinoma). The MTT (cytotoxic) data showed the inhibition growth of lung cancer cells followed pyrogallol treatment. The cell cycle of lung cancer cells was arrested in G2/M phase using flow cytometry. Using Western blot analysis, the cell cycle related proteins - cyclin B1 and Cdc25c were decreased in a time-dependent manner and the phosphorylated Cdc2 (Thr14) was increased within 4h pyrogallol treatment. Moreover, the higher cleavage of poly (ADP)-ribose polymerase (PARP), the increased of Bax concurrent with the decreased of Bcl-2 indicated that pyrogallol treatment resulted in apoptosis of lung cancer cells. The cell apoptosis was also directly demonstrated using Annexin V-FITC and TUNEL stain. Additionally, the tumoricidal effect of pyrogallol was measured using a xenograft nude mice model. After 5 weeks of pyrogallol treatment could cause the regression of tumor. Taken in vitro and in vivo studies together, these results suggest that pyrogallol can be developed as a promising anti-lung cancer drug particular for the non-small cell lung cancer (NSCLC).
Subject(s)
Antineoplastic Agents/pharmacology , Cell Cycle/drug effects , Growth Inhibitors/pharmacology , Lung Neoplasms/drug therapy , Pyrogallol/pharmacology , Adenocarcinoma/drug therapy , Animals , Antineoplastic Agents/toxicity , Carcinoma, Squamous Cell/drug therapy , Cell Cycle Proteins/drug effects , Cell Division/drug effects , Cell Line, Tumor , Cytotoxins/pharmacology , Disease Models, Animal , G2 Phase/drug effects , Growth Inhibitors/toxicity , Humans , Mice , Mice, Nude , Pyrogallol/chemistry , Pyrogallol/toxicityABSTRACT
Fissistigma oldhamii is widely used in traditional Chinese medicine to treat rheumatoid arthritis. Activation of neutrophils is a key feature of inflammatory diseases. Herein, the anti-inflammatory functions of isopedicin, a flavanone derived from F. oldhamii, and its underlying mechanisms were investigated in human neutrophils. Isopedicin potently and concentration-dependently inhibited superoxide anion (O(2)(*)(-)) production in formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP)-activated human neutrophils with an IC(50) value of 0.34+/-0.03 microM. Furthermore, isopedicin displayed no superoxide-scavenging ability, and it failed to alter subcellular NADPH oxidase activity. The inhibitory effect of isopedicin on O(2)(*)(-) production was reversed by protein kinase A (PKA) inhibitors. Moreover, isopedicin increased cAMP formation and PKA activity in FMLP-activated human neutrophils, which occurred through the inhibition of phosphodiesterase (PDE) activity but not an increase in adenylate cyclase function. In addition, isopedicin reduced FMLP-induced phosphorylation of extracellular regulated kinase and c-Jun N-terminal kinase, which was reversed by the PKA inhibitor. In contrast, isopedicin failed to alter FMLP-induced phosphorylation of p38 mitogen-activated protein kinase and calcium mobilization. In summary, these results demonstrate that inhibition of O(2)(*)(-) production in human neutrophils by isopedicin is associated with an elevation of cellular cAMP and activation of PKA through its inhibition of cAMP-specific PDE.
Subject(s)
Annonaceae , Anti-Inflammatory Agents/pharmacology , Drugs, Chinese Herbal , Flavanones/pharmacology , Neutrophils/drug effects , Superoxides/antagonists & inhibitors , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/isolation & purification , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/immunology , Cells, Cultured , Cyclic AMP/immunology , Cyclic AMP/metabolism , Cyclic AMP-Dependent Protein Kinases/immunology , Cyclic AMP-Dependent Protein Kinases/metabolism , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Enzyme Induction/drug effects , Enzyme Induction/immunology , Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors , Extracellular Signal-Regulated MAP Kinases/immunology , Flavanones/chemistry , Flavanones/immunology , Flavanones/isolation & purification , Humans , Inflammation/drug therapy , MAP Kinase Kinase 4/antagonists & inhibitors , MAP Kinase Kinase 4/immunology , Neutrophil Activation/drug effects , Neutrophil Activation/immunology , Neutrophils/enzymology , Neutrophils/immunology , Plant Stems , Superoxides/metabolismABSTRACT
Using a bioactivity-guided fractionation method, two coumarins: minumicroline acetonide (1) and epimurpaniculol senecioate (2), were isolated from the leaves of Murraya omphalocarpa Hayata (Rutaceae). Compound 1 had been previously synthesized and was now isolated from natural sources for the first time, and compound 2, possessing a negative optical rotation value, is new. The structures and their stereochemistry were fully elucidated on the basis of spectroscopic and X-ray crystallographic techniques. Both compounds 1 and 2 are active in the antiplatelet aggregation assay. Interestingly, the possible acetonide artifact 1 displayed significant antiplatelet aggregation induced not only by AA and collagen but also by platelet activating factor (PAF).
Subject(s)
Coumarins/isolation & purification , Murraya/chemistry , Plant Leaves/chemistry , Platelet Aggregation Inhibitors/isolation & purification , Animals , Arachidonic Acid/pharmacology , Blood Platelets/drug effects , Collagen/pharmacology , Coumarins/chemistry , Models, Molecular , Murraya/drug effects , Plant Leaves/drug effects , Platelet Activating Factor/pharmacology , Platelet Aggregation/drug effects , Platelet Aggregation Inhibitors/chemistry , Rabbits , Thrombin/pharmacology , X-Ray DiffractionABSTRACT
The aim of the present study was to investigate the antioxidant activity of aqueous extracts of Toona sinensis (TS; 0-100 microg/mL) and gallic acid (0-50 microg/mL), with the purified natural phenolic components evaluated using different antioxidant models. It was found that the TS extracts and gallic acid possess effective antioxidant activity against various oxidative systems in vitro, including the scavenging of free and superoxide anion radicals, reducing power, and metal chelation. However, antioxidant activity in terms of metal chelation was not observed for the gallic acid. Moreover, TS extracts and gallic acid appear to possess powerful antioxidant properties with respect to oxidative modification of human LDL induced by CuSO4, AAPH or sodium nitroprusside, as assessed by the relative electrophoretic mobility, TBARS formation, and cholesterol degradation of oxidized LDL. Furthermore, AAPH-induced oxidative hemolysis, lipid peroxidation, and decline in superoxide dismutase (SOD) activity in human erythrocytes were prevented by both the TS extracts and the gallic acid. Our findings suggest that T. sinensis may act as a chemopreventative agent, providing antioxidant properties and offering effective protection from atherogenesis.
Subject(s)
Antioxidants/pharmacology , Meliaceae/chemistry , Animals , Antioxidants/chemistry , Biphenyl Compounds , Chelating Agents/chemistry , Cholesterol, LDL/chemistry , Copper Sulfate/chemistry , Electrophoretic Mobility Shift Assay , Erythrocytes/drug effects , Free Radical Scavengers/chemistry , Gallic Acid/pharmacology , Hemolysis/drug effects , In Vitro Techniques , Lipid Peroxidation/drug effects , Nitroprusside/chemistry , Oxidation-Reduction , Picrates/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Leaves/chemistry , Superoxides/chemistry , Thiobarbituric Acid Reactive Substances/chemistryABSTRACT
Toona sinensis (T. sinensis), well known in Taiwan as a traditional Chinese medicine, has been shown to exhibit antioxidant effects. In this study, therefore, the ability of T. sinensis to induce apoptosis was studied in cultured human premyelocytic leukemia HL-60 cells. Treatment of the HL-60 cells with a variety of concentrations of the aqueous extracts of T. sinensis (TS extracts) (10-75 microg/ml) and gallic acid (5-10 microg/ml), the natural phenolic components purified from TS extracts, resulted in dose- and time-dependent sequences of events marked by apoptosis, as shown by loss of cell viability and internucleosomal DNA fragmentation. Furthermore, apoptosis in the HL-60 cells was accompanied by the release of cytochrome c, caspase 3 activation and specific proteolytic cleavage of poly (ADP-ribose) polymerase (PARP). This increase in TS extracts- and gallic acid-induced apoptosis was also associated with a reduction in the levels of Bcl-2, a potent cell-death inhibitor, and an increase in those of the Bax protein, which heterodimerizes with and thereby inhibits Bcl-2. Interestingly, TS extracts- and gallic acid-induced dose-dependent reactive oxygen species (ROS) generation in HL-60 cells. We found that catalase significantly decreased TS extracts- or gallic acid-induced cytotoxicity, DNA fragmentation, and ROS production, however, slight reduction was observed with vitamins C and E. Our results indicate that TS extracts- or gallic acid-induced HL-60 apoptotic cell death could be due to the generation of ROS, especially H(2)O(2). The data suggest that T. sinensis exerts antiproliferative action and growth inhibition on HL-60 cells through apoptosis induction, and, therefore, that it may have anticancer properties valuable for application in food and drug products.