ABSTRACT
The endocannabinoid system (ECS) is dysregulated in various liver diseases. Previously, we had shown that the major endocannabinoid 2-arachidonoyl glycerol (2-AG) promoted tumorigenesis of intrahepatic cholangiocarcinoma (ICC). However, biosynthesis regulation and clinical significance of 2-AG remain elusive. In the present study, we quantified 2-AG by gas chromatography/mass spectrometry (GC/MS) and showed that 2-AG was enriched in patients with ICC samples as well as in thioacetamide-induced orthotopic rat ICC model. Moreover, we found that diacylglycerol lipase ß (DAGLß) was the principal synthesizing enzyme of 2-AG that significantly upregulated in ICC. DAGLß promoted tumorigenesis and metastasis of ICC in vitro and in vivo and positively correlated with clinical stage and poor survival in patients with ICC. Functional studies showed that activator protein-1 (AP-1; heterodimers of c-Jun and FRA1) directly bound to the promoter and regulated transcription of DAGLß, which can be enhanced by lipopolysaccharide (LPS). miR-4516 was identified as the tumor-suppressing miRNA of ICC that can be significantly suppressed by LPS, 2-AG, or ectopic DAGLß overexpression. FRA1 and STAT3 were targets of miR-4516 and overexpression of miRNA-4516 significantly suppressed expression of FRA1, SATA3, and DAGLß. Expression of miRNA-4516 was negatively correlated with FRA1, SATA3, and DAGLß in patients with ICC samples. Our findings identify DAGLß as the principal synthesizing enzyme of 2-AG in ICC. DAGLß promotes oncogenesis and metastasis of ICC and is transcriptionally regulated by a novel AP-1/DAGLß/miR4516 feedforward circuitry.NEW & NOTEWORTHY Dysregulated endocannabinoid system (ECS) had been confirmed in various liver diseases. However, regulation and function of 2-arachidonoyl glycerol (2-AG) and diacylglycerol lipase ß (DAGLß) in intrahepatic cholangiocarcinoma (ICC) remain to be elucidated. Here, we demonstrated that 2-AG was enriched in ICC, and DAGLß was the principal synthesizing enzyme of 2-AG in ICC. DAGLß promotes tumorigenesis and metastasis in ICC via a novel activator protein-1 (AP-1)/DAGLß/miR4516 feedforward circuitry.
Subject(s)
Bile Duct Neoplasms , Cholangiocarcinoma , MicroRNAs , Rats , Animals , Transcription Factor AP-1/genetics , Endocannabinoids , Lipoprotein Lipase , Glycerol , Lipopolysaccharides , Cholangiocarcinoma/pathology , MicroRNAs/genetics , MicroRNAs/metabolism , Bile Ducts, Intrahepatic/metabolism , Bile Ducts, Intrahepatic/pathology , Bile Duct Neoplasms/metabolism , Carcinogenesis , Cell Line, TumorABSTRACT
Optical imaging provides noninvasive powerful tools not only for better understanding the physiological and pathological roles of methylglyoxal (MGO) in living systems but also for potential clinical diagnosis of MGO-related diseases, such as diabetic complications. However, so far only very few "turn-on" MGO fluorescent sensors have been developed, and they are all based on the reaction between MGO and benzenediamines. Due to the possible reactions of benzenediamines with other cellular molecules, such as NO and FA, these sensors suffer from limited selectivity and potential deactivation in cells. Herein, we report a novel MGO recognition reaction using 2-aminoacetamide. The reaction between MGO and 2-aminoacetamide was found to be highly efficient and specific, with no interference from NO and FA in particular. This reaction was used to develop the first ratiometric fluorescent probe (CMFP) for MGO. We have proven that CMFP could detect MGO at physiological concentrations in both aqueous buffer and living cells with excellent selectivity and sensitivity. Furthermore, we successfully utilized CMFP to study intracellular MGO generation routes and evaluated MGO levels of clinic blood samples from healthy and diabetic patients. These results highlight the potential utility of this probe in both basic science research and clinical diagnosis.
Subject(s)
Diabetes Complications/blood , Diabetes Mellitus/blood , Fluorescent Dyes/chemistry , Optical Imaging/methods , Pyruvaldehyde/blood , Spectrometry, Fluorescence/methods , Case-Control Studies , Cell Survival , HumansABSTRACT
Reactive sulfur species (RSS) play regulatory roles in biological systems. Many RSS are highly reactive and transient molecules, making their study difficult. RSS donors or precursors, which are used to specifically produce or deliver RSS, are useful research tools, as well as possible therapeutic agents. In this Minireview, we summarized the chemical strategies that have been used in the design of donors/precursors of hydrogen sulfide relevant RSS (including hydrogen sulfide, hydrogen polysulfides, persulfides, and S-nitroso-hydrogen sulfide). The potential problems of applying these strategies in biological settings are also discussed.
ABSTRACT
Three new diterpenoids, dumetoranes A (1) and B (2), melanocane B (3), together with four known ones including melanocane A (4), ent-15S,16-dihydroxypimar-8(14)-en-19-oic acid (5), ent-pimara-8(14),15-diene-19-oic acid (6), and ent-pimara-8(14),15-diene-19-ol (7) were obtained from the ethanol extract of the roots of Aralia dumetorum. Their structure elucidation was achieved by the methods of spectroscopic HRMS, IR, NMR, and by comparison with literature. The cytotoxicities of compounds 1-3 and 5 were assayed by in vitro MTT methods.
Subject(s)
Aralia/chemistry , Diterpenes/isolation & purification , Diterpenes/chemistry , Diterpenes/pharmacology , Magnetic Resonance Spectroscopy , Plant Roots/chemistryABSTRACT
Thionitrous acid (HSNO), the smallest S-nitrosothiol, is emerging as a potential key intermediate in cellular redox regulation linking two signaling molecules H2 S and NO. However, the chemical biology of HSNO remains poorly understood. A major hurdle is the lack of methods for selective detection of HSNO in biological systems. Herein, we report the rational design, synthesis, and evaluation of the first fluorescent probe TAP-1 for HSNO detection. TAP-1 showed high selectivity and sensitivity to HSNO in aqueous media and cells, providing a useful tool for understanding the functions of HSNO in biology.
Subject(s)
Fluorescent Dyes/metabolism , Hydrogen Sulfide/chemistry , Nitric Oxide/chemistry , S-Nitrosothiols/analysis , S-Nitrosothiols/metabolism , Fluorescent Dyes/chemistry , HeLa Cells , Humans , Molecular Structure , Oxidation-ReductionABSTRACT
Hydrogen sulfide (H2 S) is an important signaling molecule whose up- and down-regulation have specific biological consequences. Although significant advances in H2 S up-regulation, by the development of H2 S donors, have been achieved in recent years, precise H2 S down-regulation is still challenging. The lack of potent/specific inhibitors for H2 S-producing enzymes contributes to this problem. We expect the development of H2 S scavengers is an alternative approach to address this problem. Since chemical sensors and scavengers of H2 S share the same criteria, we constructed a H2 S sensor database, which summarizes key parameters of reported sensors. Data-driven analysis led to the selection of 30 potential compounds. Further evaluation of these compounds identified a group of promising scavengers, based on the sulfonyl azide template. The efficiency of these scavengers in inâ vitro and inâ vivo experiments was demonstrated.
Subject(s)
Free Radical Scavengers/chemistry , Hydrogen Sulfide/analysis , Animals , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Dose-Response Relationship, Drug , Free Radical Scavengers/pharmacology , HeLa Cells , Humans , Hydrogen Sulfide/antagonists & inhibitors , Hydrogen Sulfide/pharmacology , Methylene Blue/chemistry , Mice , Molecular Structure , Structure-Activity Relationship , Survival AnalysisABSTRACT
receptor potential canonical (TRPC) channels are presently an emerging target for airway disorders. Recent evidence has indicated that TRPC6 as a member of the TRPC family plays an important role in airway inflammation, but its precise function in bronchial epithelial cells remains unclear. The aim of this study was to investigate the role of TRPC6 in Toll-like receptor 4 (TLR4)-mediated inflammation in human bronchial epithelial cells stimulated by endotoxin [lipopolysaccharide (LPS)]. Hyp9 is a simplified phloroglucinol derivative of hyperforin that highly selectively activates TRPC6 channels. The results show that the activation of TRPC6 by Hyp9 induced the production of interleukin (IL)-8 and IL-6. LPS was also able to induce the release of IL-8 and IL-6, which was significantly aggravated by Hyp9 and reduced by knockdown of TRPC6. Treatment with LPS not only chronically induced the expression of TRPC6 mRNA and protein in a TLR4-dependent manner but also acutely increased Ca2+ influx through TRPC6 channels. In addition, LPS-induced overexpression of TRPC6 and Ca2+ influx were associated with the phosphorylation of phosphatidylinositol 3-kinase (PI3K) and Akt. Importantly, TRPC6 was required for the activation of ERK1/2, p38, and NF-κB. In conclusion, these data reveal that LPS induced the overexpression of TRPC6 and TRPC6-dependent Ca2+ influx via the TLR4/PI3K/Akt pathway resulting in Ca2+ mobilization, which subsequently promoted the activation of ERK1/2, p38, and NF-κB and the inflammatory response in bronchial epithelial cells.
Subject(s)
Bronchi/diagnostic imaging , Epithelial Cells/drug effects , Inflammation/chemically induced , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , TRPC6 Cation Channel/agonists , p38 Mitogen-Activated Protein Kinases/metabolism , Bronchi/enzymology , Calcium Signaling/drug effects , Cell Line , Cytokines/metabolism , Epithelial Cells/enzymology , Humans , Inflammation/enzymology , Inflammation/genetics , Inflammation Mediators/metabolism , NF-kappa B/metabolism , Phloroglucinol/analogs & derivatives , Phloroglucinol/pharmacology , Phosphatidylinositol 3-Kinase/metabolism , Phosphorylation , Proto-Oncogene Proteins c-akt/metabolism , TRPC6 Cation Channel/genetics , TRPC6 Cation Channel/metabolism , Terpenes/pharmacology , Toll-Like Receptor 4/agonists , Toll-Like Receptor 4/metabolismABSTRACT
Hydrogen sulphide (H2 S) serves as a vital gastric mucosal defence under acid condition. Non-steroidal anti-inflammatory drugs (NSAIDs) are among widely prescribed medications with effects of antipyresis, analgesia and anti-inflammation. However, their inappropriate use causes gastric lesions and endogenous H2 S deficiency. In this work, we reported the roles of a novel pH-controlled H2 S donor (JK-1) in NSAID-related gastric lesions. We found that JK-1 could release H2 S under mild acidic pH and increase solution pH value. Intragastrical administration of aspirin (ASP), one of NSAIDs, to mice elicited significant gastric lesions, evidenced by mucosal festering and bleeding. It also led to infiltration of inflammatory cells and resultant releases of IL-6 and TNF-α, as well as oxidative injury including myeloperoxidase (MPO) induction and GSH depletion. In addition, the ASP administration statistically inhibited H2 S generation in gastric mucosa, while up-regulated cyclooxygenase (COX)-2 and cystathionine gamma lyase (CSE) expression. Importantly, these adverse effects of ASP were prevented by the intragastrical pre-administration of JK-1. However, JK-1 alone did not markedly alter the property of mouse stomachs. Furthermore, in vitro cellular experiments showed the exposure of gastric mucosal epithelial (GES-1) cells to HClO, imitating MPO-driven oxidative injury, decreased cell viability, increased apoptotic rate and damaged mitochondrial membrane potential, which were reversed by pre-treatment with JK-1. In conclusion, JK-1 was proved to be an acid-sensitive H2 S donor and could attenuate ASP-related gastric lesions through reconstruction of endogenous gastric defence. This work indicates the possible treatment of adverse effects of NSAIDs with pH-controlled H2 S donors in the future.
Subject(s)
Aspirin/toxicity , Gastric Mucosa/drug effects , Hydrogen Sulfide/metabolism , Organothiophosphonates/pharmacology , Protective Agents/pharmacology , Animals , Anti-Inflammatory Agents, Non-Steroidal/toxicity , Cell Line , Cyclooxygenase 2/metabolism , Cystathionine gamma-Lyase/metabolism , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Gastric Mucosa/metabolism , Gastric Mucosa/pathology , Humans , Hydrogen-Ion Concentration , Interleukin-6/metabolism , Male , Mice , Molecular Structure , Organothiophosphonates/chemistry , Organothiophosphonates/metabolism , Protective Agents/chemistry , Protective Agents/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
BACKGROUND/AIM: Accumulation of advanced glycation end products (AGEs) is a major cause of diabetes mellitus (DM) skin complications. Methylglyoxal (MGO), a reactive dicarbonyl compound, is a crucial intermediate of AGEs generation. N-acetyl-L-cysteine (NAC), an active ingredient of some medicines, can induce endogenous GSH and hydrogen sulfide generation, and set off a condensation reaction with MGO. However, there is rare evidence to show NAC can alleviate DM-induced skin injury through inhibition of AGEs generation or toxicity. The present study aimed to observe the effects of NAC on MGO-induced inflammatory injury and investigate the roles of AGEs and its receptor (RAGE) in NAC's dermal protection in human HaCaT keratinocytes. METHODS: The cells were exposed to MGO to simulate a high MGO status in diabetic blood or tissues. The content of AGEs in serum or cell medium was measured with ELISA. The protective effects of NAC against MGO-induce injury were evaluated by administration before MGO one hour, in virtue of cell viability, mitochondrial membrane potential, inflammation reaction, nuclear factor (NF)-κB activation, matrix metalloproteinase (MMP)-9 expression, as well as cellular behavioral function. RESULTS: We found the AGEs levels of patients with DM were elevated comparing with healthy volunteers. The in vitro AGEs generation was also able to be enhanced by the exposure of HaCaT cells to MGO, which reduced dose-dependently cellular viability, damaged mitochondrial function, triggered secretion of interleukin (IL)-6 and IL-8, activated NF-κB and upregulated MMP-9 expression. Furthermore, the exposure caused cellular adhesion and migration dysfunction, as well as collagen type I inhibition. Importantly, before the exposure to MGO, the preconditioning with NAC significantly attenuated MGO-induced AGEs generation, improved cellular viability and mitochondrial function, partially reversed the overexpression of proinflammatory factors and MMP-9, as well as the activation of NF-κB. Lastly, NAC blocked MGO-induced RAGE upregulation, and inhibition of RAGE with its neutralizing antibody significantly alleviated MGO-induced NF-κB activation, MMP-9 upregulation and inflammatory injury in HaCaT cells. CONCLUSION: The present work indicates the administration of NAC can prevent MGO-induced dermal inflammatory injury through inhibition of AGEs/RAGE signal, which may provide a basal support for the treatment of diabetic skin complications with NAC-containing medicines in the future.
Subject(s)
Acetylcysteine/pharmacology , Glycation End Products, Advanced/analysis , Protective Agents/pharmacology , Pyruvaldehyde/pharmacology , Receptor for Advanced Glycation End Products/analysis , Up-Regulation/drug effects , Aged , Case-Control Studies , Cell Adhesion/drug effects , Cell Line , Cell Movement/drug effects , Cell Survival/drug effects , Diabetes Mellitus, Type 2/metabolism , Diabetes Mellitus, Type 2/pathology , Female , Glycation End Products, Advanced/blood , Humans , Interleukin-6/analysis , Interleukin-8/analysis , Male , Matrix Metalloproteinase 9/metabolism , Membrane Potential, Mitochondrial/drug effects , Middle Aged , Mitochondria/drug effects , Mitochondria/metabolism , Receptor for Advanced Glycation End Products/bloodABSTRACT
Hydrogen sulfide (H2S) is a critical signaling molecule that regulates many physiological and/or pathological processes. Modulation of H2S levels could have potential therapeutic value. In this work, we report the rational design, synthesis, and biological evaluation of a class of phosphonamidothioate-based H2S-releasing agents (i.e., H2S donors). A novel pH-dependent intramolecular cyclization was employed to promote H2S release from the donors. These water-soluble compounds showed slow, controllable, and pH-sensitive production of H2S in aqueous solutions. The donors also showed significant cytoprotective effects in cellular models of oxidative damage. Most importantly, the donors were found to exhibit potent cardioprotective effects in an in vivo murine model of myocardial ischemia-reperfusion (MI/R) injury through a H2S-related mechanism.
Subject(s)
Disease Models, Animal , Hydrogen Sulfide/metabolism , Myocardial Reperfusion Injury/metabolism , Animals , Hydrogen-Ion Concentration , MiceABSTRACT
Ammonium tetrathiomolybdate (TTM) was found to be a slow hydrogen sulfide (H2S) releasing agent. Its H2S generation capability in aqueous solutions was confirmed by UV-vis and fluorescence assays. TTM also showed H2S-like cytoprotective effects in hydrogen peroxide (H2O2)-induced oxidative damage in HaCaT cells.
Subject(s)
Hydrogen Sulfide/chemistry , Molybdenum/chemistry , Water/chemistry , Cell Line , Cell Survival/drug effects , Dose-Response Relationship, Drug , Humans , Hydrogen Peroxide/pharmacology , Molecular Structure , Molybdenum/pharmacology , Oxidation-Reduction/drug effects , Oxidative Stress/drug effects , Solubility/drug effects , Structure-Activity RelationshipABSTRACT
Five new diterpenoids (1-5) were isolated from the roots of Aralia melanocarpa, together with four known compounds, 7ß-hydroxy-ent-pimara-8(14),15-diene-19-oic acid (6), 18-norpimara-8(14),15-dien-4-ol (7), ent-16ßH,17-isovalerate-kauran-19-oic acid (8), and ent-16α,17-dihydroxykauran-19-oic acid (9). Based on the MS, IR, and NMR spectral analysis, the structures of the five new diterpenoids (1-5) were elucidated. The cytotoxic activities of compounds 1-9 were assayed, and compounds 1 and 2 showed cytotoxicity in four cancer cell lines with IC50s from 4.2 to 8.2 µM.
Subject(s)
Antineoplastic Agents, Phytogenic/isolation & purification , Aralia/chemistry , Diterpenes/isolation & purification , Antineoplastic Agents, Phytogenic/chemistry , Cell Line , Diterpenes/chemistry , Diterpenes/pharmacology , Drug Screening Assays, Antitumor , Molecular Structure , Plant Roots/chemistryABSTRACT
In this study, we investigated the protective effect of the antioxidant N-acetyl-L-cysteine (NAC) on the lung inflammation caused by ozone (O3) exposure in mice. Thirty-two C57BL/6 mice were randomly divided into control group, O3 group, O3+NAC group and NAC group. Mice were exposed to O3 (1.0 ppm) or fresh air for 3 h on the day 1, day 3 and day 5, respectively. NAC (100 mg/kg) was intraperitoneally applied to the mice 1 h before each exposure. At 24 h after the 3-time exposure, the alveolar wall structure was severely damaged and the infiltrated inflammatory cells were apparent perivascularly and peribronchiolarly. Significant increases in the total white blood cell count, macrophage, lymphocyte and neutrophil counts, as well as total protein concentration were observed in the bronchoalveolar lavage fluid (BALF) (P < 0.05). The IL-6, IL-8 (P < 0.01) and MDA levels (P < 0.05) in the lung homogenates were elevated coherently. Administration of NAC could attenuate the alveolar wall structure damage induced by O3 exposure and reduce the amount of infiltrated inflammatory cells, total and differential leukocyte counts (P < 0.05), as well as the IL-6, IL-8 (P < 0.01) and MDA release (P < 0.05). Western blotting results showed that the O3 exposure up-regulated the p38 MAPK and NF-κB p65 protein expression in the lung tissue of mice (P < 0.05), which could be alleviated by NAC (P < 0.05). These results indicated that NAC could protect against O3-induced pulmonary inflammation in mice. The beneficial effect of NAC might be related with the p38 MAPK and NF-κB p65 signal pathway.
Subject(s)
Pneumonia , Acetylcysteine , Animals , Antioxidants , Bronchoalveolar Lavage Fluid , Interleukin-6 , Lung , Mice , Mice, Inbred C57BL , NF-kappa B , Neutrophils , OzoneABSTRACT
BACKGROUND/AIM: Delayed wound healing is a common skin complication of diabetes, which is associated with keratinocyte injury and dysfunction. Levels of methylglyoxal (MGO), an α-dicarbonyl compound, are elevated in diabetic skin tissue and plasma, while levels of hydrogen sulfide (H2S), a critical gaseous signaling molecule, are reduced. Interestingly, the gas has shown dermal protection in our previous study. To date, there is no evidence demonstrating whether MGO affects keratinocyte viability and function or H2S donation abolishes these effects and improves MGO-related impairment of wound healing. The current study was conducted to examine the effects of MGO on the injury and function in human skin keratinocytes and then to evaluate the protective action of a novel H2S-releasing molecule. METHODS: An N-mercapto-based H2S donor (NSHD)-1 was synthesized and its ability to release H2S was observed in cell medium and cells, respectively. HaCaT cells, a cell line of human skin keratinocyte, were exposed to MGO to establish an in vitro diabetic wound healing model. NSHD-1 was added to the cells before MGO exposure and the improvement of cell function was observed in respect of cellular viability, apoptosis, oxidative stress, mitochondrial membrane potential (MMP) and behavioral function. RESULTS: Treatment with MGO decreased cell viability, induced cellular apoptosis, increased intracellular reactive oxygen species (ROS) content and depressed MMP in HaCaT cells. The treatment also damaged cell behavioral function, characterized by decreased cellular adhesion and migration. The synthesized H2S-releasing molecule, NSHD-1, was able to increase H2S levels in both cell medium and cells. Importantly, pretreatment with NSHD-1 inhibited MGO-induced decreases in cell viability and MMP, increases in apoptosis and ROS accumulation in HaCaT cells. The pretreatment was also able to improve adhesion and migration function. CONCLUSION: These results demonstrate that the novel synthesized H2S donor is able to protect human skin keratinocytes against MGO-induced injury and behavior dysfunction. We believe that more reasonable H2S-releasing molecules will bring relief to patients suffering from delayed wound healing in diabetes mellitus in the future.
Subject(s)
Hydrogen Sulfide/pharmacology , Keratinocytes/drug effects , Protective Agents/pharmacology , Pyruvaldehyde/pharmacology , Skin Diseases/chemically induced , Skin Diseases/drug therapy , Skin/drug effects , Apoptosis/drug effects , Cell Line , Cell Survival/drug effects , Humans , Keratinocytes/metabolism , Membrane Potential, Mitochondrial/drug effects , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , Skin Diseases/metabolism , Wound Healing/drug effectsABSTRACT
BACKGROUND: Cardiac hypertrophy is a compensatory stage of the heart in response to stress such as pressure overload (PO), which can develop into heart failure (HF) if left untreated. Resveratrol has been reported to prevent the development of hypertrophy and contractile dysfunction induced by PO. However, other studies found that resveratrol treatment for a longer period of time failed to regress cardiac hypertrophy. The aim of this study is to determine the timing of resveratrol treatment to achieve antihypertrophic effect and investigate whether resveratrol prevents the development of HF through preservation of myocardium structure and modulation of Ca(2+) handling proteins. METHODS: To generate rats with cardiac hypertrophy, male Sprague-Dawley rats were subjected to PO (aortic banding procedure) for 4 weeks. Sham-operated animals served as controls. Rats with cardiac hypertrophy were given resveratrol (4 mg/kg/day) for 4, 6, and 8 weeks, respectively. Histological and echocardiographic analysis and transmission electron microscopy were performed to assess cardiac structure and function. The levels of Ca(2+) handling proteins were measured by western blot analysis. RESULTS: Histological analysis showed that resveratrol treatment regressed developed cardiac hypertrophy at 8 and 10 weeks postsurgery, but not at 12 weeks. However, resveratrol strongly and continuously prevented the development of cardiac dysfunction and dilation of cardiac chamber as evaluated by echocardiography and H&E staining of heart cross-sections. In addition, PO-induced cardiac fibrosis was completely inhibited by resveratrol treatment. Resveratrol markedly prevented the disrupted myocardium but partially rescued mitochondrial abnormality in banded rats. Moreover, resveratrol prevented the alteration of Ca(2+) handling proteins induced by aortic banding, including downregulation of sarcoplasmic reticulum Ca(2+) ATPase 2 (SERCA2) and ryanodine receptor 2 (RyR2), hypophosphorylated phospholamban (PLB), upregulation of Na(+)/Ca(2+)-exchangers (NCX1) and increased expression and phosphorylation of Ca(2+)/calmodulin -dependent protein kinase II (CaMKII). Moreover, resveratrol alleviated the decreased SERCA activity induced by aortic banding. CONCLUSIONS: Resveratrol effectively prevented the transition from compensatory to decompensatory stage of cardiac hypertrophy induced by PO, but this effect is dependent on the timing of treatment. We suggest that resveratrol may exert beneficial effects on cardiac hypertrophy through protection of cardiac structure and modulation of Ca(2+) handling proteins.
Subject(s)
Calcium/metabolism , Heart/drug effects , Stilbenes/pharmacology , Animals , Heart/physiopathology , Male , Pressure , Rats , Rats, Sprague-Dawley , ResveratrolABSTRACT
Accumulating evidence indicates that angiotensin-(1-7) [Ang-(1-7)] offers protective effects against ischemia-reperfusion (I/R) induced arrhythmias and contractile dysfunction, which are related to disturbances of intracellular calcium homeostasis. However, whether or not Ang-(1-7) regulates intracellular calcium in I/R is not clear. To shed light on this issue, we carried out studies with a cellular model of simulated I/R in isolated rat ventricular myocytes and measured calcium transients using laser scanning confocal microscopy. Our results showed that Ang-(1-7) had no effects on the calcium transient in myocytes superfused with normal solution; however, in myocytes of simulated I/R, Ang-(1-7) significantly attenuated the increased diastolic intracellular Ca during reperfusion, restored the decreased peak Ca of calcium transient during ischemia, and reversed the decreased amplitude of calcium transient throughout the I/R periods. Additionally, Ang-(1-7) significantly suppressed the reactive oxygen species production in I/R, especially during the ischemic phase. These data indicated that Ang-(1-7) affords significant cytoprotective effects through directly improving calcium homeostasis independent of its anti-oxidative action. Most notably, the effects of Ang-(1-7) on intracellular Ca dynamics manifests only in the diseased states, that is, I/R. This unique property suggests that upregulation of Ang-(1-7) expression and/or activation of the Ang-(1-7)/Mas signaling cascade is a highly desirable strategy for the treatment of myocardial impairment induced by I/R.
Subject(s)
Angiotensin I/pharmacology , Calcium/metabolism , Myocardial Reperfusion Injury/drug therapy , Peptide Fragments/pharmacology , Animals , Cytoprotection/drug effects , Homeostasis/drug effects , Male , Microscopy, Confocal , Myocardial Reperfusion Injury/physiopathology , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/metabolism , Rats , Rats, Sprague-DawleyABSTRACT
Four new clerodane diterpenes, namely sagittatayunnanosides A-D (1-4), were isolated from the roots of Tinospora sagittata var. yunnanensis, together with two known compounds, tinospinoside C (5) and tinospinoside E (6). The structures of the four new compounds were well elucidated by extensive analyses of the MS, IR, and 1D and 2D NMR data. The cytotoxic and antifouling activities of compounds 1-6 were evaluated.
Subject(s)
Diterpenes, Clerodane/isolation & purification , Tinospora/chemistry , Cell Line, Tumor , Diterpenes, Clerodane/chemistry , Humans , Magnetic Resonance Spectroscopy/methods , Spectrometry, Mass, Electrospray Ionization , Spectrophotometry, InfraredABSTRACT
10 accessions of tetraploid wheat were radiated with 100 Gy (60)Co γ -ray. The germination energy, germination rate, special characters (secondary tillering, stalk with wax powder, and dwarf), meiotic process, and high-molecular-weight glutenin subunits (HMW-GSs) were observed. Different species has different radiation sensibility. With 1 seed germinated (5%), T. dicoccum (PI434999) is the most sensitive to this dose of radiation. With a seed germination rate of 35% and 40%, this dose also affected T. polonicum (As304) and T. carthlicum (As293). Two mutant dwarf plants, T. turgidum (As2255) 253-10 and T. polonicum (As302) 224-14, were detected. Abnormal chromosome pairings were observed in pollen mother cells of both T. dicoccoides (As835) 237-9 and T. dicoccoides (As838) 239-8 with HMW-GS 1Ax silent in seeds from them. Compared with the unirradiated seed of T. polonicum (As304) CK, a novel HMW-GS was detected in seed of T. polonicum (As304) 230-7 and its electrophoretic mobility was between 1By8 and 1Dy12 which were the HMW-GSs of Chinese Spring. These mutant materials would be resources for wheat breeding.
Subject(s)
Cobalt Radioisotopes , Gamma Rays/adverse effects , Mutation/radiation effects , Tetraploidy , Triticum/genetics , Triticum/radiation effects , Germination/genetics , Germination/radiation effects , Mitosis , Phenotype , Radiation DosageABSTRACT
BACKGROUND/AIM: The abnormal activation of the AKT/GSK3ß signal pathway in lymphocytes from systemic lupus erythematosus (SLE) patients plays an important role in the pathogenesis of the disease. Recently Hydrogen sulfide (H2S) has been recognized as a crucial gaseous signaling molecule, involved in regulation of cell proliferation. However, the role of H2S in regulating the abnormal activation of lymphocytes from SLE patients has not been established. This study was conducted to investigate the effect of H2S on lymphocytes and to explore the mechanisms involved. METHODS: The lymphocytes were isolated from SLE patients with or without renal disease and healthy controls. The cells were treated as indicated in each experiment. Cell viability was analyzed by CCK-8. Cell cycle distribution was determined by flow cytometry. Western blot was used to detect the expression of phosphorylated AKT (ser473), GSK3ß (ser9) and CDK2, p27(Kip1) and p21(WAF1/CIP1). RESULTS: Our findings showed that proliferation of lymphocytes was stimulated following treatment with NaHS (a H2S donor) at low NaHS concentrations (<1mM) but inhibited at high NaHS concentrations (>2mM). Similar results were observed using GYY4137, which is a slow-releasing H2S donor. Pretreatment of lymphocytes from SLE patients with NaHS at high concentrations prior to exposure to phytohemagglutinin (PHA) significantly attenuated proliferation, evidenced by decrease in cell viability and S phase distribution of cell cycle. Pretreatment with NaHS decreased PHA-induced expression of CDK2, phosphorylation levels of AKT (ser473) and GSK3ß (ser9) and increased the expression of p27(Kip1) and p21(WAF1/CIP1). Moreover, pretreatment with NaHS blunted the stimulation of SLE lymphocyte proliferation by GSK3ß inhibitor lithium chloride. CONCLUSION: These results demonstrate that H2S inhibits the abnormal activation of lymphocytes from SLE patients throuqh the AKT/GSK3ß signal pathway.
Subject(s)
Glycogen Synthase Kinase 3/metabolism , Hydrogen Sulfide/pharmacology , Lymphocytes/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Cyclin-Dependent Kinase 2/metabolism , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Glycogen Synthase Kinase 3/antagonists & inhibitors , Glycogen Synthase Kinase 3 beta , Humans , Lithium Chloride/pharmacology , Lupus Erythematosus, Systemic/metabolism , Lupus Erythematosus, Systemic/pathology , Lymphocytes/cytology , Lymphocytes/metabolism , Morpholines/pharmacology , Organothiophosphorus Compounds/pharmacology , Phosphorylation , Phytohemagglutinins/pharmacology , S Phase Cell Cycle Checkpoints/drug effects , Signal Transduction/drug effectsABSTRACT
BACKGROUND: Cerebral atherosclerosis (CA) is a chronic disease caused by multiple infarcts and atrophy causing nerve degenerative syndrome. Ginkgo Folium (GF) and Forsythiae Fructus (FF) have shown positive effects on vascular protection, but their relationship with CA is unclear. This study aimed to identify the potential CA targets and mechanisms of action of GF-FF, using network pharmacology. OBJECTIVE: This study used network pharmacology and molecular docking to examine the potential targets and pharmacological mechanism of GF-FF on CA. METHODS: Using the traditional Chinese medicine systems pharmacology database and analysis platform, components were screened and corresponding targets were predicted using boundary values and Swiss Target Prediction. Using Cytoscape 3.8.0, a network was established between GF-FF components and CA targets. We extracted disease genes and constructed a network of targets based on the protein-protein interaction networks functional enrichment analysis database. Using Metascape, the Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes of the enriched targets were determined. AutoDock Vina was used to perform molecular docking. RESULTS: Twenty-three active ingredients of GF-FF were confirmed to treat CA, covering 109 targets, of which 48 were CA-related. Luteolin, bicuculline, sesamin, kaempferol, quercetin, and ginkgolide B were the vital active compounds, and EGFR, CYP2E1, CREB1, CYP19A1, PTGS2, PPARG, PPARA, ESR1, MMP9, MAPK14, MAPK8, and PLG were the major targets. The molecular docking showed that these compounds and targets exhibited good intercalation. These 48 protein targets produced effects on CA by modulating pathways such as "apoptosis-multiple species," "IL-17 signaling pathway," and "relaxin signaling pathway." CONCLUSIONS: As predicted by network pharmacology, GF-FF exerts anti-tumor effects through multiple components and targets for treatment of CA, providing new clinical ideas for CA treatment.