ABSTRACT
Curcumin (diferuloylmethane) is a major component of turmeric, which is isolated from the rhizomes of Curcuma longa L. from the family Zingiberaceae. It is used as a dietary pigment for curry and in traditional Indian medicine for its anti-inflammatory and attenuating pain effects. This study aimed to evaluate the beneficial effects of curcumin in a rat model of diabetic neuropathic pain. Additionally, we investigated the involvement of the phosphorylated form of c-Jun N-terminal kinase (pJNK) located in the neurons and astrocytes of the dorsal root ganglion (DRG). To induce diabetic neuropathic pain in rats, 50 mg/kg of streptozotocin (STZ) was intraperitoneally injected. After 4 weeks, rats were administered the vehicle, 10 mg/kg/day curcumin, or 50 mg/kg/day curcumin orally for 4 consecutive weeks. One day after the final drug administration, we performed behavioral tests to measure responses of rats to mechanical, heat, cold, and acetone-induced cold stimuli. After behavioral tests, pJNK expression in the DRG was evaluated using western blot assay and immunohistochemistry. Curcumin treatment for 4 consecutive weeks in STZ-induced diabetic neuropathic pain rats improved behavioral responses to mechanical, cold, and thermal stimuli. Increased pJNK expression in the astrocytes and neurons of the DRG in STZ-induced diabetic neuropathic pain rats was reduced by curcumin treatment for 4 consecutive weeks. We suggest that curcumin can be an option for the treatment of diabetes-related neuropathic pain, and one of the mechanisms that underlie the action of curcumin may involve pJNK expression in the astrocytes and neurons of the DRG.
Subject(s)
Astrocytes/drug effects , Curcumin/therapeutic use , Diabetic Neuropathies/drug therapy , JNK Mitogen-Activated Protein Kinases/drug effects , Animals , Curcumin/pharmacology , Diabetes Mellitus, Experimental , Disease Models, Animal , Ganglia, Spinal/metabolism , Male , Rats , Rats, Sprague-DawleyABSTRACT
Sanguinarine (SAN), a quaternary benzophenanthridine alkaloid extracted from the root of Papaveraceae plants, has shown antitumour effects in multiple cancer cells. However, the therapeutic effects and the underlying mechanisms of SAN in gastric cancer (GC) remain elusive. In this study, the in vitro proliferation inhibition effect of SAN in GC cells was determined using CCK-8 assay, the in vivo antitumor effect of SAN was evaluated in mice with xenotransplanted tumor. The mechanism underlying the antitumor activity of SAN was explored by gene microarray assay and bioinformatics analysis. The levels of differentially expressed miRNAs and target genes were verified by real-time RT-PCR and immunohistochemistry. SAN inhibited the proliferation of BGC-823 cells in a concentration-dependent manner in vitro and in vivo. The miR-96-5p and miR-29c-3p were significantly upregulated in untreated BGC-823 cells and significantly downregulated in SAN treated cells. The mRNA and protein expression of their target gene MAP4K4 were upregulated in SAN treated xenotransplanted tumors, and pMEK4 and pJNK1 proteins in the MAPK/JNK signaling pathway were also upregulated by SAN. These indicate that SAN may inhibit the proliferation of BGC-823 cells through the inhibition of miR-96-5p and miR-29c-3p expression, and subsequent activation of the MAPK/JNK signaling pathway.
Subject(s)
Antineoplastic Agents/pharmacology , Benzophenanthridines/pharmacology , Isoquinolines/pharmacology , JNK Mitogen-Activated Protein Kinases/drug effects , MicroRNAs/biosynthesis , Stomach Neoplasms/drug therapy , Animals , Cell Line, Tumor , Cell Proliferation/drug effects , Down-Regulation/drug effects , Gene Expression Regulation, Neoplastic , HeLa Cells , Hep G2 Cells , Humans , Intracellular Signaling Peptides and Proteins/drug effects , MAP Kinase Signaling System/drug effects , MCF-7 Cells , Mice , Mice, Inbred BALB C , Mice, Nude , Papaveraceae/chemistry , Protein Serine-Threonine Kinases/drug effects , RNA, Messenger/biosynthesis , Stomach Neoplasms/genetics , Xenograft Model Antitumor AssaysABSTRACT
Nowadays, medicinal plants have been widely used everywhere to provide essential care for many disorders including diabetes. Recent reports assumed that the antidiabetic activities of pomegranate aril juice (PAJ) may be ascribed to its punicalagin (PCG). Therefore, the present study evaluated and compared the antidiabetic activities of PAJ and its PCG, and monitored some mechanisms of their actions in streptozotocin-nicotinamide (STZ-NA) type 2 diabetic rats. STZ-NA diabetic rats were given, orally/daily, PAJ (100 or 300 mg/kg body weight, containing 2.6 and 7.8 mg of PCG/kg body weight, respectively), pure PCG (2.6 or 7.8 mg/kg body weight), or distilled water (vehicle) for 6 weeks. PAJ (especially at the high dose) alleviated significantly (P < 0.05-0.001) most signs of type 2 diabetes including body-weight loss, insulin resistance (IR) and hyperglycemia through decreasing serum tumor necrosis factor-α concentration and the expression of hepatic c-Jun N-terminal kinase, and increasing the skeletal muscle weight and the expression of hepatic insulin receptor substrate-1 in STZ-NA diabetic rats. Also, it decreased significantly (P < 0.001) the oxidative liver injury in STZ-NA diabetic rats through decreasing the hepatic lipid peroxidation and nitric oxide production, and improving the hepatic antioxidant defense system. Although the low dose of PCG induced some modulation in STZ-NA diabetic rats, the high dose of PCG did not show any valuable antidiabetic activity, but induced many side effects. In conclusion, PAJ was safer and more effective than pure PCG in alleviating IR and oxidative liver injury in STZ-NA diabetic rats.
Subject(s)
Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/metabolism , Hydrolyzable Tannins/administration & dosage , Hydrolyzable Tannins/therapeutic use , Insulin Resistance , Liver/drug effects , Liver/pathology , Niacinamide/administration & dosage , Pomegranate/metabolism , Streptozocin/administration & dosage , Animals , Antioxidants/metabolism , Blood Glucose/metabolism , Body Weight/drug effects , Hydrolyzable Tannins/metabolism , Hyperglycemia/drug therapy , Insulin Receptor Substrate Proteins/drug effects , JNK Mitogen-Activated Protein Kinases/drug effects , Lipid Peroxidation/drug effects , Liver/metabolism , Male , Muscle, Skeletal/drug effects , Niacinamide/metabolism , Nitric Oxide/metabolism , Rats , Streptozocin/metabolism , Tumor Necrosis Factor-alpha/drug effectsABSTRACT
BACKGROUND: Skeletal muscle atrophy is a striking example of the multiple changes in the physiological state of humans and animals induced by microgravity. Previous studies have shown that a blood circulation disorder may be a cause of this atrophy, and traditional Chinese medicine has been regarded as a potential countermeasure to reverse the atrophy in China. This study was carried out to test the effects of Xuefuzhuyu capsules (XFZY) on the skeletal muscle atrophy induced by simulated microgravity. METHODS: The mass and cross-sectional area of the soleus muscle were compared in rats treated with XFZY that were hindlimb unloaded for 30 d (XFZY-TS group), untreated rats that were hindlimb unloaded for 30 d (TS group), and control rats (CON group). The expression and phosphorylation levels of key proteins of the sarcoplasmic reticulum stress system were also measured. RESULTS: Treatment with XFZY attenuated the loss of muscle mass and cross-sectional area induced by hindlimb unloading. Western blot analysis showed that the phosphatidyl-inositol-3-kinase/phospho-Akt (PI3K/p-Akt) pathways were down-regulated after 30 d in the TS group compared with the CON group. This effect was partly reversed by XFZY. Hindlimb unloading increased the expression of glucose-regulated protein 78 (GRP78), cytosine-cytosine-adenosine-adenosine-thymidine/enhancer-binding protein homologous protein (CHOP), C-Jun N-terminal kinase (JNK), and Caspase 12. Treatment with XFZY alleviated this increased protein expression. DISCUSSION: Our results suggest that XFZY could partially reverse the effects of hindlimb unloading on muscle atrophy and perhaps target the sarcoplasmic reticulum stress system, possibly through the GRP78-CHOP-JNK-Caspase 12 pathway.Zhang S, Yuan M, Cheng C, Xia D, Wu S. Chinese herbal medicine effects on muscle atrophy induced by simulated microgravity. Aerosp Med Hum Perform. 2018; 89(10):883-888.
Subject(s)
Drugs, Chinese Herbal/pharmacology , Hindlimb Suspension , Muscle, Skeletal/drug effects , Muscular Atrophy/pathology , Weightlessness Simulation , Animals , Caspase 12/drug effects , Caspase 12/metabolism , Endoplasmic Reticulum Chaperone BiP , Heat-Shock Proteins/drug effects , Heat-Shock Proteins/metabolism , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , MAP Kinase Signaling System , Male , Muscle, Skeletal/metabolism , Muscle, Skeletal/pathology , Muscular Atrophy/metabolism , Phosphatidylinositol 3-Kinase/drug effects , Phosphatidylinositol 3-Kinase/metabolism , Phosphorylation/drug effects , Proto-Oncogene Proteins c-akt/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Rats , Rats, Sprague-Dawley , Signal Transduction , Transcription Factor CHOP/drug effects , Transcription Factor CHOP/metabolismABSTRACT
Dysregulated human monocytes/macrophages can synthesize and secrete matrix metalloproteinases (MMPs), which play important roles in the progression of sepsis. In this study, we investigated the effects and mechanism of a novel histone deacetylase (HDAC8) inhibitor, (E)-N-hydroxy-4-methoxy-2-(biphenyl-4-yl)cinnamide (WK2-16), on MMP-9 production and activation in stimulated human monocytic THP-1 cells. Our results demonstrated that the acetylation level of structural maintenance of chromosomes 3 (SMC3) was up-regulated by WK2-16 in THP-1 cells. Consistently, an in vitro enzyme study demonstrated that WK2-16 selectively inhibited HDAC8 activity. Moreover, the WK2-16 concentration dependently suppressed MMP-9-mediated gelatinolysis induced by tumor necrosis factor-α (TNF-α) or lipopolysaccharide (LPS). Additionally, WK2-16 significantly inhibited both MMP-9 protein and mRNA expression without cellular toxicity. Nevertheless, WK2-16 suppressed the extracellular levels of interleukin (IL)-6 from LPS-stimulated THP-1 cells. For the signaling studies, WK2-16 had no effect on LPS/TLR4 downstream signaling pathways, such as the NF-κB and ERK/JNK/P38 MAPK pathways. On the other hand, WK2-16 enhanced the recruitment of acetylated Yin Yang 1 (YY1) with HDAC1. Finally, in vivo studies indicated that WK2-16 could reduce the serum levels of TNF-α and IL-6 in endotoxemic mice. These results suggested that HDAC8 inhibition might provide a novel therapeutic strategy of hypercytokinemia in sepsis.
Subject(s)
Cytokines/drug effects , Cytokines/metabolism , Histone Deacetylase Inhibitors/pharmacology , Histone Deacetylases/drug effects , Histone Deacetylases/metabolism , Lipopolysaccharides/pharmacology , Matrix Metalloproteinase 9/drug effects , Matrix Metalloproteinase 9/metabolism , Repressor Proteins/drug effects , Repressor Proteins/metabolism , Acetylation , Animals , Cell Cycle Proteins/metabolism , Cell Survival/drug effects , Chondroitin Sulfate Proteoglycans/metabolism , Chromosomal Proteins, Non-Histone/metabolism , Cyclooxygenase 2/drug effects , Down-Regulation , Endotoxemia , Histone Deacetylase 1/drug effects , Humans , Interleukin-6 , JNK Mitogen-Activated Protein Kinases/drug effects , MAP Kinase Signaling System/drug effects , Male , Mice , Mice, Inbred C57BL , Monocytes/metabolism , NF-kappa B/metabolism , RNA, Messenger/biosynthesis , RNA, Messenger/drug effects , Sepsis/drug therapy , Signal Transduction/drug effects , THP-1 Cells/drug effects , Tubulin/metabolism , Tumor Necrosis Factor-alpha/metabolism , Tumor Necrosis Factor-alpha/pharmacology , YY1 Transcription Factor/metabolism , p38 Mitogen-Activated Protein Kinases/drug effectsABSTRACT
BACKGROUND At present, the treatment of coxsackievirus-induced myocarditis remains difficult. Berberine (BBR), an isoquinoline alkaloid isolated from traditional medicine herbs, exhibits significant anti-viral efficacy against various viruses. However, the underlying mechanism by which BBR controls CVB3 infection has not yet been reported. The purpose of this study was to investigate the anti-viral efficacy of BBR against CVB3 infection and its mechanism. MATERIAL AND METHODS In our experiments, the protein levels of VP1 and MAPKs signal pathway were measured by Western blot. The mRNA level of VP1 was measured by RT-PCR. The virus titers were determined by TCID50 assay. RESULTS We found that BBR treatment significantly decreased CVB3 replication in HeLa cells. In addition, the BBR treatment reduced the phosphorylation levels of JNK and p38 MAPK upon CVB3 infection in both HeLa cells and primary rat myocardial cells. CONCLUSIONS Taken together, these results suggest that BBR inhibits CVB3 replication through the suppression of JNK and p38 MAPK activation, shedding new light on the investigation of therapeutic strategies against CVB3-induced viral myocarditis.
Subject(s)
Berberine/metabolism , Berberine/therapeutic use , Enterovirus B, Human/drug effects , Virus Replication/drug effects , Animals , Antiviral Agents/pharmacology , Cell Culture Techniques , Coxsackievirus Infections , HeLa Cells , Humans , JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Myocarditis/drug therapy , Myocytes, Cardiac/metabolism , Phosphorylation , Primary Cell Culture , Rats , Receptors, Virus/metabolism , Signal Transduction/drug effects , p38 Mitogen-Activated Protein Kinases/drug effects , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
This study aimed to investigate the effects of total alkaloids from Nelumbinis Plumula (NPA) on insulin resistance (IR) of high-fat diet- (HFD-) induced nonalcoholic fatty liver disease (NAFLD). Rats were fed with HFD for 8 weeks to induce NAFLD. Then, the effect of NPA on ameliorating IR in HFD-induced NAFLD was evaluated. Fasting serum insulin was determined using an enzyme-linked immunosorbent assay (ELISA) kit for insulin following the manufacturer's protocol. Some inflammatory cytokines such as tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) were determined using ELISA kits to assess the inflammatory burden in rats. The results showed that HFD could induce a significant increase in blood glucose and IR in rats. However, rats treated with NPA (400 or 600 mg/kg) showed improved IR and reduction in serum inflammatory cytokines TNF-α and IL-6. Further investigation indicated that NPA could inhibit IR by restoring the insulin receptor substrate-1 (IRS-1) and suppressing the expression of c-Jun N-terminal kinase (JNK) phosphorylation. The present results supported the view that the pathogenesis of NAFLD was complex with inflammation, together with increasing serum glucose and IR. Also, JNK and IRS phosphorylation were suggested for their involvement in the modulating of IR during NAFLD progression. Therefore, NPA may serve as a potential natural remedy against IR in NAFLD.
Subject(s)
Alkaloids/pharmacology , Blood Glucose/drug effects , Diet, High-Fat , Insulin Resistance , Nelumbo , Non-alcoholic Fatty Liver Disease/metabolism , Plant Extracts/pharmacology , Seeds , Animals , Blood Glucose/metabolism , Insulin/metabolism , Insulin Receptor Substrate Proteins/drug effects , Insulin Receptor Substrate Proteins/metabolism , Interleukin-6/immunology , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Male , Non-alcoholic Fatty Liver Disease/immunology , Phosphoproteins/drug effects , Phosphoproteins/metabolism , Rats , Rats, Wistar , Tumor Necrosis Factor-alpha/drug effects , Tumor Necrosis Factor-alpha/immunologyABSTRACT
OBJECTIVE: Obesity-induced inflammation plays a pivotal role in the pathogenesis of insulin resistance and type 2 diabetes. Xanthoangelol (XA) and 4-hydroxyderrcin (4-HD), phytochemicals extracted from Angelica keiskei, have been reported to possess various biological properties. Whether XA and 4-HD alleviate obesity-induced inflammation and inflammation-induced adipocyte dysfunction was investigated. METHODS: For the in vitro study, a co-culture system composed of macrophages and adipocytes and macrophages stimulated with conditioned medium derived from fully differentiated adipocytes was conducted. For the in vivo study, mice were fed a high-fat diet supplemented with XA for 14 weeks. RESULTS: XA and 4-HD suppressed inflammatory factors in co-culture system. Moreover, treatment of RAW macrophages with XA and 4-HD moderated the suppression of uncoupling protein 1 promoter activity and gene expression in C3H10T1/2 adipocytes, which was induced by conditioned medium derived from LPS-stimulated RAW macrophages. Also, XA and 4-HD inhibited c-Jun N-terminal kinase phosphorylation, nuclear factor-κB, and activator protein 1, the last two being transcription activators in activated macrophages. Furthermore, in mice fed the high-fat diet, XA reduced inflammatory factors within the white adipose tissue. CONCLUSIONS: These results suggest that XA and 4-HD might be promising phytochemicals to suppress obesity-induced inflammation and inflammation-induced adipocyte dysfunction.
Subject(s)
Angelica/chemistry , Chalcone/analogs & derivatives , Obesity/drug therapy , Phytotherapy , Plant Extracts/pharmacology , Adipocytes/drug effects , Animals , Cell Differentiation/drug effects , Chalcone/pharmacology , Coculture Techniques , Culture Media, Conditioned , Diet, High-Fat , Inflammation/drug therapy , Inflammation/etiology , JNK Mitogen-Activated Protein Kinases/drug effects , Macrophages/drug effects , Male , Mice , Mice, Inbred C57BL , NF-kappa B/drug effects , Obesity/complications , Obesity/physiopathology , Phosphorylation/drug effects , Transcription Factor AP-1/drug effectsABSTRACT
This study investigated the pathways involved in the effect of green tea epigallocatechin gallate (EGCG) on mitogenesis in BeWo, JEG-3, and JAR placental choriocarcinoma cells. EGCG inhibited cell proliferation in dose-dependent and time-dependent manners, as indicated by the number of cells and incorporation of bromodeoxyuridine (BrdU). A catechin-specific effect of green tea was evident; EGCG was more effective than epicatechin, epicatechin gallate, and epigallocatechin in suppressing cell growth. When all three of the mitogen-activated protein kinase (MAPK) subfamilies, i.e., ERK, p38, and JNK, were examined, EGCG significantly increased levels of phospho-ERK1/2 (pERK1/2) and phospho-p38 (pp38) and did not alter the total protein levels of ERK1/2, p38 MAPK, JNK, and phospho-JNK. EGCG-induced increases in the levels of pERK1/2 and pp38 proteins were prevented by pre-treatment with specific inhibitors of ERK1/2 MAPK and p38 MAPK, respectively. These inhibitors also suppressed EGCG-induced decreases in both cell number and BrdU incorporation. Moreover, pre-treatment with an AMP-activated protein kinase (AMPK) inhibitor prevented the actions of EGCG on proliferation and AMPK phosphorylation. These data suggest that EGCG mediates choriocarcinoma cell growth via the AMPK, ERK, and p38 pathways, but not JNK pathway.
Subject(s)
Catechin/analogs & derivatives , Cell Proliferation/drug effects , Choriocarcinoma/pathology , Tea , Uterine Neoplasms/pathology , AMP-Activated Protein Kinases/drug effects , AMP-Activated Protein Kinases/metabolism , Antimitotic Agents , Catechin/pharmacology , Cell Line, Tumor , Cell Proliferation/physiology , Extracellular Signal-Regulated MAP Kinases/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , Female , Humans , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , MAP Kinase Signaling System/drug effects , MAP Kinase Signaling System/physiology , Phosphorylation , Pregnancy , p38 Mitogen-Activated Protein Kinases/drug effects , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
BACKGROUND: Clinical use of chemotherapeutic drug, cisplatin is limited by its toxicity and drug resistance. Therefore, efforts continue for the discovery of novel combination therapies with cisplatin, to increase efficacy and reduce its toxicity. Here, we screened 16 medicinal plant extracts from Northeast part of India and found that leaf extract of Zanthoxylum armatum DC. (ZALE) induced cytotoxicity as well as an effect on the increasing of the efficiency of chemotherapeutic drugs (cisplatin, mitomycin C and camptothecin). This work shows detail molecular mechanism of anti-cancer activity of ZALE and its potential for combined treatment regimens to enhance the apoptotic response of chemotherapeutic drugs. RESULTS: ZALE induced cytotoxicity, nuclear blebbing and DNA fragmentation in HeLA cells suggesting apoptosis induction in human cervical cell line. However, the apoptosis induced was independent of caspase 3 activation and poly ADP ribose polymerase (PARP) cleavage. Further, ZALE activated Mitogen-activated protein kinases (MAPK) pathway as revealed by increased phosphorylation of extracellular-signal-regulated kinases (ERK), p38 and c-Jun N-terminal kinase (JNK). Inhibition of ERK activation but not p38 or JNK completely blocked the ZALE induced apoptosis suggesting an ERK dependent apoptosis. Moreover, ZALE generated DNA double strand breaks as suggested by the induction γH2AX foci formation. Interestingly, pretreatment of certain cancer cell lines with ZALE, sensitized the cancer cells to cisplatin and other chemotherapeutic drugs. Enhanced caspase activation was observed in the synergistic interaction among chemotherapeutic drugs and ZALE. CONCLUSION: Purification and identification of the bio-active molecules from the ZALE or as a complementary treatment for a sequential treatment of ZALE with chemotherapeutic drugs might be a new challenger to open a new therapeutic window for the novel anti-cancer treatment.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Cisplatin/pharmacology , Plant Extracts/pharmacology , Zanthoxylum/chemistry , Apoptosis/drug effects , Enzyme Activation/drug effects , HeLa Cells , Humans , JNK Mitogen-Activated Protein Kinases/drug effects , Mitogen-Activated Protein Kinases/drug effectsABSTRACT
BACKGROUND: Clinical use of chemotherapeutic drug, cisplatin is limited by its toxicity and drug resistance. Therefore, efforts continue for the discovery of novel combination therapies with cisplatin, to increase efficacy and reduce its toxicity. Here, we screened 16 medicinal plant extracts from Northeast part of India and found that leaf extract of Zanthoxylum armatum DC. (ZALE) induced cytotoxicity as well as an effect on the increasing of the efficiency of chemotherapeutic drugs (cisplatin, mitomycin C and camptothecin). This work shows detail molecular mechanism of anti-cancer activity of ZALE and its potential for combined treatment regimens to enhance the apoptotic response of chemotherapeutic drugs. RESULTS: ZALE induced cytotoxicity, nuclear blebbing and DNA fragmentation in HeLA cells suggesting apoptosis induction in human cervical cell line. However, the apoptosis induced was independent of caspase 3 activation and poly ADP ribose polymerase (PARP) cleavage. Further, ZALE activated Mitogen-activated protein kinases (MAPK) pathway as revealed by increased phosphorylation of extracellular-signal-regulated kinases (ERK), p38 and c-Jun N-ter-minal kinase (JNK). Inhibition of ERK activation but not p38 or JNK completely blocked the ZALE induced apoptosis suggesting an ERK dependent apoptosis. Moreover, ZALE generated DNA double strand breaks as suggested by the induction γH2AX foci formation. Interestingly, pretreatment of certain cancer cell lines with ZALE, sensitized the cancer cells to cisplatin and other chemotherapeutic drugs. Enhanced caspase activation was observed in the synergistic interaction among chemotherapeutic drugs and ZALE. CONCLUSION: Purification and identification of the bio-active molecules from the ZALE or as a complementary treatment for a sequential treatment of ZALE with chemotherapeutic drugs might be a new challenger to open a new therapeutic window for the novel anti-cancer treatment.
Subject(s)
Humans , Plant Extracts/pharmacology , Cisplatin/pharmacology , Zanthoxylum/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , HeLa Cells , Apoptosis/drug effects , Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/drug effects , Enzyme Activation/drug effectsABSTRACT
Magnolol, a neolignan from the traditional medicinal plant Magnolia obovata, has been shown to possess neuroprotective, anti-inflammatory, anticancer and anti-angiogenic activities. However, the precise mechanism of the anti-angiogenic activity of magnolol remains to be elucidated. In the present study, the anti-angiogenic effect of magnolol was evaluated in mouse embryonic stem (mES)/embryoid body (EB)-derived endothelial-like cells. The endothelial-like cells were obtained by differentiation from mES/EB cells. Magnolol (20 µM) significantly suppressed the transcriptional and translational expression of platelet endothelial cell adhesion molecule (PECAM), an endothelial biomarker, in mES/EB-derived endothelial-like cells. To further understand the molecular mechanism of the suppression of PECAM expression, signaling pathways were analyzed in the mES/EB-derived endothelial-like cells. Magnolol induced the generation of reactive oxygen species (ROS) by mitochondria, a process that was associated with the induction of apoptosis as determined by positive Annexin V staining and the activation of cleaved caspase-3. The involvement of ROS generation by magnolol was confirmed by treatment with an antioxidant, N-acetyl-cysteine (NAC). NAC inhibited the magnolol-mediated induction of ROS generation and suppression of PECAM expression. In addition, magnolol suppressed the activation of MAPKs (ERK, JNK and p38) and the PI3K/AKT/mTOR signaling pathway in mES/EB-derived endothelial-like cells. Taken together, these findings demonstrate for the first time that the anti-angiogenic activity of magnolol may be associated with ROS-mediated apoptosis and the suppression of the PI3K/AKT/mTOR signaling pathway in mES/EB-derived endothelial-like cells.
Subject(s)
Angiogenesis Inhibitors/pharmacology , Apoptosis/drug effects , Biphenyl Compounds/pharmacology , Lignans/pharmacology , MAP Kinase Signaling System/drug effects , Neovascularization, Pathologic/drug therapy , Reactive Oxygen Species/metabolism , Acetylcysteine/metabolism , Animals , Caspase 3/biosynthesis , Caspase 3/drug effects , Caspase 3/metabolism , Cell Differentiation , Cell Line , Embryonic Stem Cells/cytology , Embryonic Stem Cells/drug effects , Embryonic Stem Cells/metabolism , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Enzyme Activation/drug effects , Extracellular Signal-Regulated MAP Kinases/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Membrane Potential, Mitochondrial/drug effects , Mice , Mitochondria/drug effects , Mitochondria/metabolism , Nitric Oxide Synthase/antagonists & inhibitors , Phosphatidylinositol 3-Kinases/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Platelet Endothelial Cell Adhesion Molecule-1/biosynthesis , Platelet Endothelial Cell Adhesion Molecule-1/drug effects , Proto-Oncogene Proteins c-akt/drug effects , Proto-Oncogene Proteins c-akt/metabolism , TOR Serine-Threonine Kinases/drug effects , TOR Serine-Threonine Kinases/metabolism , p38 Mitogen-Activated Protein Kinases/drug effects , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
Oxidative stress increases with age and is postulated to be a major causal factor for sarcopenia in aging. Here, we examined whether the administration of a cystine-based antioxidant (F1) can alleviate/delay age-specific changes in skeletal muscles. C57BL6 male mice aged 17 months (middle aged) were fed with normal diet with or without supplementation of F1 (3 mg/kg food) for 6 months. Compared with young (5 months old) mice old mice exhibited increased markers of oxidative stress, inflammation, and muscle cell apoptosis and decreased muscle weight. These age-related changes were further associated with inactivation of adenosine-5'-monophosphate-activated protein kinase (AMPK), increased lipogenesis, activation of c-Jun NH2-terminal kinase, and decreased expression of Delta 1, phospho-Akt, and proliferating cell nuclear antigen in aged skeletal muscle. Such alterations were significantly prevented by F1. These results demonstrate the beneficial effects of F1 to attenuate loss of muscle mass associated with aging.
Subject(s)
Aging , Antioxidants/therapeutic use , Apoptosis/drug effects , Cystine/therapeutic use , Muscle, Skeletal/drug effects , Oxidative Stress/drug effects , Sarcopenia/drug therapy , AMP-Activated Protein Kinases/drug effects , Animals , Antioxidants/metabolism , Cystine/metabolism , Disease Models, Animal , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Male , Mice , Mice, Inbred C57BL , Treatment OutcomeABSTRACT
BACKGROUND: Nonalcoholic fatty liver disease is a chronic metabolic disorder with significant impact on cardiovascular and liver mortality. AIMS: In this study, we examined the effects of silibinin on liver and myocardium injury in an experimental model of nonalcoholic fatty liver disease. METHODS: A four-week daily dose of silibinin (20 mg/kg i.p.) was administrated to db/db mice fed a methionine-choline deficient diet. Hepatic and myocardial histology, oxidative stress and inflammatory cytokines were evaluated. RESULTS: Silibinin administration decreased HOMA-IR, serum ALT and markedly improved hepatic and myocardial damage. Silibinin reduced isoprostanes, 8-deoxyguanosine and nitrites/nitrates in the liver and in the heart of db/db fed the methionine-choline deficient diet, whereas glutathione levels were restored to lean mice levels in both tissues. Consistently, liver mitochondrial respiratory chain activity was significantly impaired in untreated mice and was completely restored in silibinin-treated animals. TNF-α was increased whereas IL-6 was decreased both in the liver and heart of db/db fed methionine-choline deficient diet. Silibinin reversed heart TNF-α and IL-6 expression to control mice levels. Indeed, liver JNK phosphorylation was reduced to control levels in treated animals. CONCLUSIONS: This study demonstrates a combined effectiveness of silibinin on improving liver and myocardial injury in experimental nonalcoholic fatty liver disease.
Subject(s)
Antioxidants/therapeutic use , Fatty Liver/drug therapy , Fatty Liver/pathology , Myocardium/pathology , Silymarin/therapeutic use , Alanine Transaminase/blood , Alanine Transaminase/drug effects , Analysis of Variance , Animals , Antioxidants/pharmacology , Choline Deficiency/metabolism , Cytokines/drug effects , Cytokines/genetics , Cytokines/metabolism , Diet , Fatty Liver/blood , Gene Expression/drug effects , Glutathione/drug effects , Glutathione/metabolism , Insulin Resistance , Isoprostanes/metabolism , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Liver/metabolism , Liver/pathology , Male , Methionine/deficiency , Mice , Myocardium/metabolism , Nitrates/metabolism , Nitrites/metabolism , Non-alcoholic Fatty Liver Disease , Oxidative Stress/drug effects , Phosphorylation/drug effects , Silybin , Silymarin/pharmacology , Statistics, NonparametricABSTRACT
BACKGROUND AND OBJECTIVE: Host modulatory agents directed at inhibiting specific proinflammatory mediators could be beneficial in terms of attenuating periodontal disease progression and potentially enhancing therapeutic responses. The aim of this study was to investigate whether daidzein could modulate the production inflammatory mediators in macrophages stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen implicated in periodontal disease, and to delineate underlying mechanisms of action. MATERIAL AND METHODS: LPS was extracted from P. intermedia ATCC 25611 cells by the standard hot phenol-water method. The amounts of nitric oxide (NO) and interleukin-6 (IL-6) secreted into the culture medium were assayed. A real-time PCR was performed to quantify inducible nitric oxide synthase (iNOS) and IL-6 mRNA expression. We used immunoblot analysis to characterize iNOS protein expression, phosphrylation of c-Jun N-terminal kinase (JNK) and p38, degradation of inhibitory κB-α (IκB-α), nuclear translocation of nuclear factor-κB (NF-κB) subunits and phosphorylation of signal transducer and activator of transcription 1 (STAT1). The DNA-binding activity of NF-κB was assessed by using ELISA-based kits. RESULTS: Daidzein significantly inhibited the production of NO and IL-6, as well as their mRNA expression, in P. intermedia LPS-treated RAW264.7 cells. The JNK and p38 pathways were not involved in the regulation of LPS-induced NO and IL-6 release by daidzein. Daidzein inhibited the degradation of IκB-α induced by P. intermedia LPS. In addition, daidzein suppressed NF-κB transcriptional activity via regulation of the nuclear translocation and DNA-binding activity of NF-κB p50 subunit and blocked STAT1 phosphorylation. CONCLUSION: Although additional studies are required to dissect the molecular mechanism of action, our results suggest that daidzein could be a promising agent for treating inflammatory periodontal disease. Further research in animal models of periodontitis is necessary to better evaluate the potential of daidzein as a novel therapeutic agent to treat periodontal disease.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Growth Inhibitors/pharmacology , Isoflavones/pharmacology , Lipopolysaccharides/pharmacology , Macrophages/drug effects , Phytoestrogens/pharmacology , Prevotella intermedia , Animals , Bacteriological Techniques , Cell Culture Techniques , Cell Line , I-kappa B Kinase/drug effects , Inflammation Mediators/antagonists & inhibitors , Interleukin-6/antagonists & inhibitors , JNK Mitogen-Activated Protein Kinases/drug effects , Janus Kinase 2/drug effects , Mice , NF-kappa B/drug effects , NF-kappa B p50 Subunit/drug effects , Nitric Oxide Synthase Type II/drug effects , Phosphorylation , STAT1 Transcription Factor/drug effects , Transcription Factor RelA/drug effects , p38 Mitogen-Activated Protein Kinases/drug effectsABSTRACT
Hydroalcoholic extract of Solidago chilensis (Sc) is employed in popular medicine to treat inflammatory disease. The low-grade proinflammatory state and the activation of serine/threonine kinases in adipose tissue, like c-jun kinase (JNK) and IKK, and transcription factors, have an important role in obesity-associated insulin resistance. The aim of this study was to further investigate the effects of the Sc extract on glucose homeostasis in diet-induced obesity mice. Male Swiss mice were randomized to three groups: a control group (C) fed with standard laboratory chow; a group with an experimental high-fat diet (HFD); and a group fed with a high-fat (45% kcal from fat) diet + extract of Sc (via intraperitoneal, 3 mg/kg) (ScHFD). The dietary treatment lasted for eight weeks. Subsequently, the expression and phosphorylation of proteins of interest in the liver, hypothalamus and skeletal muscle were evaluated by Western blot analysis. Body weight, epididymal fat pad mass and liver triglycerides were higher in HFD than in control mice, but these parameters were reduced by intraperitoneal administration of the extracts (3 mg/kg) to the HFD group. AKT phosphorylation stimulated by insulin in the liver, hypothalamus and skeletal muscle was higher in ScHFD as compared with HFD mice. Additionally, liver expression of phosphoenolpyruvate carboxykinase (PEPCK) and fatty acid synthase were lower in ScHFD as compared with HFD mice. Nuclear factor κB, p-IκB and p-JNK levels were higher in HFD when compared with control mice, but they were lowered by treatment with extract (ScHFD). In addition, in db/db mice, Sc extract also improved liver AKT phosphorylation stimulated by insulin and reduced PEPCK expression. The data presented herein show that Sc improves AKT activation. This effect may be promoted by reduction of the proinflammatory pathway in the liver and hypothalamus. Therefore, systemic action of the Sc components may contribute to improve obesity-associated pathophysiology.
Subject(s)
I-kappa B Proteins/metabolism , Insulin Resistance , JNK Mitogen-Activated Protein Kinases/metabolism , Obesity/metabolism , Plant Extracts/therapeutic use , Solidago , Animals , Glucose/analysis , Glucose Tolerance Test , I-kappa B Proteins/drug effects , JNK Mitogen-Activated Protein Kinases/drug effects , Liver/chemistry , Liver/metabolism , Liver Glycogen/analysis , Male , Mice , Signal Transduction/drug effects , Triglycerides/analysisABSTRACT
Strong anti-neoplastic anthracyclines like daunorubicin (DNR) and doxorubicin (DOX) have high efficacy against systemic neoplasm and solid tumours. However, clinically, they cause chronic cardiomyopathy and congestive heart failure. Red palm oil (RPO) supplementation can protect the heart against ischemic injury. We therefore hypothesize that supplementation with RPO during chemotherapy may protect the heart. Control rats received a standard diet, and the experimental group received RPO in addition for 4 weeks. Each group was subsequently injected with either saline or DNR over a 12-day period towards the end of 4 weeks. Hearts were excised and perfused on a working heart system. Functional parameters were measured. Tissue samples were collected for analysis of mRNA and protein levels. DNR + RPO increased aortic output by 25% (p < 0.05) compared with DNR only. Furthermore, DNR treatment significantly reduced tissue mRNA levels of superoxide dismutase 1 (SOD1) and nitric oxide synthase 1 (NOS1) compared with untreated controls. Protein expression of SOD1 followed the same pattern as mRNA levels. NOS1 protein levels were significantly increased in DNR treated rats when compared with untreated controls. In addition, DNR increased phosphorylation of p38 and Jun N-terminal kinase compared with untreated controls, whereas DNR + RPO completely counteracted this activation. DNR + RPO significantly up regulated the protein extracellular signal-regulated kinase 1 level compared with DNR only. In this model of DNR treatment, RPO is associated with stabilization of important antioxidant enzymes such NOS and SOD, and inhibition of the 'stress' induced mitogen-activated protein kinase pathways. Dietary RPO also maintained function, similar to control, in DNR treated hearts.
Subject(s)
Anthracyclines/adverse effects , Antibiotics, Antineoplastic/adverse effects , Heart/physiology , Plant Oils/pharmacology , Animals , Antioxidants/pharmacology , Aorta/drug effects , Aorta/physiology , Daunorubicin/adverse effects , Dietary Supplements , Heart/drug effects , Heart Function Tests , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Male , Mitogen-Activated Protein Kinase 3/drug effects , Mitogen-Activated Protein Kinase 3/metabolism , Nitric Oxide Synthase/drug effects , Nitric Oxide Synthase/genetics , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type I , Palm Oil , Phosphorylation/drug effects , Protective Agents/pharmacology , RNA, Messenger , Rats , Rats, Wistar , Signal Transduction , Superoxide Dismutase/drug effects , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolism , Superoxide Dismutase-1 , p38 Mitogen-Activated Protein Kinases/drug effects , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
The role of individual supplements necessary for the long-term self-renewal of embryonic stem (ES) cells is poorly characterized in feeder/serum-free culture systems. This study sought to characterize the relationship between the effects of glucose on ES cell proliferation and fibronectin (FN) synthesis, and to assess the mechanisms responsible for these cellular effects of glucose. Treatment of the two ES cells (ES-E14TG2a and ES-R1) with 25 mM glucose (high glucose) increased the expression levels of FN mRNA and protein. In addition, high glucose and ANG II synergistically increased FN expression level, which coincident with data showing that high glucose increased the mRNA expression of angiotensin II (ANG II) type 1 receptor (AT(1)R), angiotensinogen, and FN, but not ANG II type 2 receptor. High glucose also increased the intracellular calcium (Ca(2+)) concentration and pan-protein kinase C (PKC) phosphorylation. Inhibition of the Ca(2+)/PKC pathway blocked high glucose-induced FN expression. High glucose or ANG II also synergistically increased transforming growth factor-beta1 (TGF-beta(1)) expression, while pretreatment with losartan abolished the high glucose-induced increase in TGF-beta(1) production. Moreover, TGF-beta(1)-specific small interfering RNA inhibited high glucose-induced FN expression and c-Jun N-terminal kinase (JNK) activation. The JNK inhibitor SP600125 blocked high glucose-induced FN expression and inhibited cell cycle regulatory protein expression induced by high glucose or TGF-beta(1). In this study, inhibition of AT(1)R, Ca(2+)/PKC, TGF-beta(1), JNK, FN receptor blocked the high glucose-induced DNA synthesis, increased the cell population in S phase, and the number of cells. It is concluded that high glucose increases FN synthesis through the ANG II or TGF-beta1 pathways, which in part mediates proliferation of mouse ES cells.
Subject(s)
Angiotensin II/metabolism , Cell Proliferation/drug effects , Embryonic Stem Cells/metabolism , Fibronectins/biosynthesis , Glucose/metabolism , Transforming Growth Factor beta1/metabolism , Angiotensin II/pharmacology , Angiotensin II Type 1 Receptor Blockers/pharmacology , Angiotensinogen/genetics , Animals , Calcium Signaling/drug effects , Calcium Signaling/physiology , Cell Count , DNA Replication/drug effects , DNA Replication/genetics , Embryonic Stem Cells/drug effects , Fibronectins/drug effects , Fibronectins/genetics , Glucose/pharmacology , Hyperglycemia/genetics , Hyperglycemia/metabolism , Hyperglycemia/physiopathology , JNK Mitogen-Activated Protein Kinases/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Losartan/pharmacology , Mice , Phosphorylation/drug effects , Protein Kinase C/drug effects , Protein Kinase C/metabolism , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Receptor, Angiotensin, Type 1/genetics , Signal Transduction/drug effects , Signal Transduction/physiology , Transforming Growth Factor beta1/drug effects , Transforming Growth Factor beta1/geneticsABSTRACT
The IGF axis is critical for the regulation of apoptosis in many human cancer cell lines. Recently, potent anti-tumorigenic effects of pomegranate juice and extracts have been reported. Consequently, pomegranate has potential not only as a treatment but also as a preventative measure against certain types of cancer, including prostate. In this study, we investigated the relationship between pomegranate-induced apoptosis in human prostate cancer cells and the IGF/IGFBP system. Treatment of LAPC4 prostate cancer cells with 10microg/ml POMx, a highly potent pomegranate extract prepared from skin and arils minus seeds and standardized to ellagitannin content (37% punicalagins by HPLC), resulted in inhibition of cell proliferation and induction of apoptosis. Interestingly, co-treatment with POMx and IGFBP-3 revealed synergistic stimulation of apoptosis and additive inhibition of cell growth. Western blot analysis revealed that treatment with POMx or POMx/IGFBP-3 combination resulted in increased JNK phosphorylation, and decreased Akt and mTOR activation, consistent with a growth inhibitory, pro-apoptotic function. We also investigated the relationship between IGF-1 and pomegranate-induced apoptosis in 22RV1 prostate cancer cells. Co-treatment with 100ng/ml IGF-1 completely blocked apoptosis induction by POMx. In contrast, IGF-I failed to inhibit POMx-induced apoptosis in R(-) cells, suggesting the importance of IGF-IR. POMx-treatment decreased Igf1 mRNA expression in a dose-dependent manner indicating that its actions also involve tumor-specific suppression of IGF-1. These studies revealed novel interactions between the IGF system and pomegranate-induced apoptosis.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis , Insulin-Like Growth Factor Binding Protein 3/metabolism , Insulin-Like Growth Factor I/metabolism , Lythraceae/chemistry , Plant Extracts/pharmacology , Prostatic Neoplasms/metabolism , Antineoplastic Agents, Phytogenic/chemistry , Cell Line, Tumor , Cell Proliferation/drug effects , Humans , Hydrolyzable Tannins/analysis , Hydrolyzable Tannins/pharmacology , Intracellular Signaling Peptides and Proteins/drug effects , JNK Mitogen-Activated Protein Kinases/drug effects , Male , Plant Extracts/chemistry , Protein Serine-Threonine Kinases/drug effects , TOR Serine-Threonine KinasesABSTRACT
Sedum telephium ssp. maximum is a medicinal plant that possesses anti-inflammatory, analgesic and keratolytic properties. We investigated the anti-inflammatory activity of its methanolic extract (STME) in rat peritoneal macrophages (MPhis) stimulated with lipopolysaccharide from Salmonella enteritidis. After stimulation with 10 microg/ml of LPS, MPhis were coincubated with different doses of STME (8, 16 and 32 microg/ml) or RPMI medium alone using different times of incubation. STME reduced levels of tumor necrosis factor-alpha, both mRNA and its protein, and significantly decreased IL-1beta and IL-6 production. Moreover, STME inhibited inducible nitric oxide synthase expression and blunted nitrite release and inhibited both extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinase activation in lipopolysaccharide-stimulated MPhis. Data show that STME might be useful as a potential anti-inflammatory agent.