ABSTRACT
BACKGROUND: Increasing global overweight and obesity rates not only increase the prevalence of myocardial infarction (MI), but also exacerbate ischemic injury and result in worsened prognosis. Currently, there are no drugs that can reverse myocardial damage once MI has occurred, therefore discovering drugs that can potentially limit the extent of ischemic damage to the myocardium is critical. Resveratrol is a polyphenol known for its antioxidant properties, however whether prolonged daily intake of resveratrol during obesity can protect against MI-induced damage remains unexplored. METHODS: We established murine models of obesity via high-fat/high-fructose diet, along with daily administrations of resveratrol or vehicle, then performed surgical MI to examine the effects and mechanisms of resveratrol in protecting against myocardial ischemic injury. RESULTS: Daily administration of resveratrol in obese mice robustly protected against myocardial ischemic injury and improved post-MI cardiac function. Resveratrol strongly inhibited oxidative and DNA damage via activating SIRT3/FOXO3a-dependent antioxidant enzymes following MI, which were completely prevented upon administration of 3-TYP, a selective SIRT3 inhibitor. Hence, the cardioprotective effects of prolonged resveratrol intake in protecting obese mice against myocardial ischemic injury was due to reestablishment of intracellular redox homeostasis through activation of SIRT3/FOXO3a signaling pathway. CONCLUSION: Our findings provide important new evidence that supports the daily intake of resveratrol, especially in those overweight or obese, which can robustly decrease the extent of ischemic damage following MI. Our study therefore provides new mechanistic insight and suggests the therapeutic potential of resveratrol as an invaluable drug in the treatment of ischemic heart diseases.
Subject(s)
Forkhead Box Protein O3 , Homeostasis , Mice, Inbred C57BL , Mice, Obese , Obesity , Oxidation-Reduction , Resveratrol , Signal Transduction , Sirtuin 3 , Animals , Resveratrol/pharmacology , Signal Transduction/drug effects , Sirtuin 3/metabolism , Male , Oxidation-Reduction/drug effects , Obesity/drug therapy , Obesity/metabolism , Obesity/complications , Forkhead Box Protein O3/metabolism , Homeostasis/drug effects , Mice , Antioxidants/pharmacology , Myocardial Ischemia/metabolism , Myocardial Ischemia/drug therapy , Oxidative Stress/drug effects , Diet, High-Fat/adverse effects , Myocardial Infarction/prevention & control , Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Myocardial Infarction/drug therapy , Cardiotonic Agents/pharmacology , Stilbenes/pharmacology , Stilbenes/therapeutic useABSTRACT
Type 2 diabetes (T2D) is a metabolic disorder that causes numerous complications including impaired wound healing and poses a significant challenge for the management of diabetic patients. Epigallocatechin-3-gallate (EGCG) is a natural polyphenol that exhibits anti-inflammatory and anti-oxidative benefits in skin wounds, however, the direct effect of EGCG on epidermal keratinocytes, the primary cells required for re-epithelialization in wound healing remains unknown. Our study aims to examine the underlying mechanisms of EGCG's ability to promote re-epithelialization and wound healing in T2D-induced wounds. Murine models of wound healing in T2D were established via feeding high-fat high-fructose diet (HFFD) and the creation of full-thickness wounds. Mice were administered daily with EGCG or vehicle to examine the wound healing response and underlying molecular mechanisms of EGCG's protective effects. Systemic administration of EGCG in T2D mice robustly accelerated the wound healing response following injury. EGCG induced nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2) and promoted cytokeratin 16 (K16) expression to activate epidermal keratinocytes and robustly promoted re-epithelialization of wounds in diabetic mice. Further, EGCG demonstrated high binding affinity with Kelch-like ECH-associated protein 1 (KEAP1), thereby inhibiting KEAP1-mediated degradation of NRF2. Our findings provide important evidence that EGCG accelerates the wound healing response in diabetic mice by activating epidermal keratinocytes, thereby promoting re-epithelialization of wounds via K16/NRF2/KEAP1 signaling axis. These mechanistic insights into the protective effects of EGCG further suggest its therapeutic potential as a promising drug for treating chronic wounds in T2D.
Subject(s)
Catechin/analogs & derivatives , Diabetes Mellitus, Experimental , Diabetes Mellitus, Type 2 , Humans , Mice , Animals , Re-Epithelialization , Kelch-Like ECH-Associated Protein 1/metabolism , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/metabolism , NF-E2-Related Factor 2/metabolism , Keratinocytes , Wound HealingABSTRACT
BACKGROUND AND AIMS: Anti-hypertensive agents are one of the most frequently used drugs worldwide. However, no blood pressure-lowering strategy is superior to placebo with respect to survival in diabetic hypertensive patients. Previous findings show that Wnt co-receptors LDL receptor-related proteins 5 and 6 (LRP5/6) can directly bind to several G protein-coupled receptors (GPCRs). Because angiotensin II type 1 receptor (AT1R) is the most important GPCR in regulating hypertension, this study examines the possible mechanistic association between LRP5/6 and their binding protein Dickkopf-1 (DKK1) and activation of the AT1R and further hypothesizes that the LRP5/6-GPCR interaction may affect hypertension and potentiate cardiac impairment in the setting of diabetes. METHODS: The roles of serum DKK1 and DKK1-LRP5/6 signalling in diabetic injuries were investigated in human and diabetic mice. RESULTS: Blood pressure up-regulation positively correlated with serum DKK1 elevations in humans. Notably, LRP5/6 physically and functionally interacted with AT1R. The loss of membrane LRP5/6 caused by injection of a recombinant DKK1 protein or conditional LRP5/6 deletions resulted in AT1R activation and hypertension, as well as ß-arrestin1 activation and cardiac impairment, possibly because of multiple GPCR alterations. Importantly, unlike commonly used anti-hypertensive agents, administration of the anti-DKK1 neutralizing antibody effectively prevented diabetic cardiac impairment in mice. CONCLUSIONS: These findings establish a novel DKK1-LRP5/6-GPCR pathway in inducing diabetic injuries and may resolve the long-standing conundrum as to why elevated blood DKK1 has deleterious effects. Thus, monitoring and therapeutic elimination of blood DKK1 may be a promising strategy to attenuate diabetic injuries.
Subject(s)
Diabetes Mellitus, Experimental , Diabetic Cardiomyopathies , Hypertension , Receptors, LDL , Animals , Humans , Mice , Antihypertensive Agents , Diabetic Cardiomyopathies/prevention & control , Hypertension/prevention & control , Receptors, LDL/antagonists & inhibitorsABSTRACT
BACKGROUND: Pathological cardiac hypertrophy is a hallmark of various cardiovascular diseases (CVD) including chronic heart failure (HF) and an important target for the treatment of these diseases. Aberrant activation of Angiotensin II (Ang II)/AT1R signaling pathway is one of the main triggers of cardiac hypertrophy, which further gives rise to excessive inflammation that is mediated by the key transcription factor NF-κB. Resveratrol (REV) is a natural polyphenol with multiple anti-inflammatory and anti-oxidative effects, however the ability of REV in preventing Ang II-induced cardiac hypertrophy in combination with NF-κB signaling activation remains unclear. METHODS: Murine models of cardiac hypertrophy was conducted via implantation of Ang II osmotic pumps. Primary neonatal rat cardiomyocyte and heart tissues were examined to determine the effect and underlying mechanism of REV in preventing Ang II-induced cardiac hypertrophy. RESULTS: Administrations of REV significantly prevented Ang II-induced cardiac hypertrophy, as well as robustly attenuated Ang II-induced cardiac fibrosis, and cardiac dysfunction. Furthermore, REV not only directly prevented Ang II/AT1R signal transductions, but also prevented Ang II-induced expressions of pro-inflammatory cytokines and activation of NF-κB signaling pathway. CONCLUSIONS: Our study provides important new mechanistic insight into the cardioprotective effects of REV in preventing Ang II-induced cardiac hypertrophy via inhibiting adverse NF-κB signaling activation. Our findings further suggest the therapeutic potential of REV as a promising drug for the treatment of cardiac hypertrophy and heart failure.
Subject(s)
Heart Failure , NF-kappa B , Rats , Mice , Animals , NF-kappa B/metabolism , Resveratrol/adverse effects , Angiotensin II/pharmacology , Signal Transduction , Cardiomegaly/chemically induced , Cardiomegaly/drug therapy , Cardiomegaly/metabolism , Myocytes, Cardiac , Heart Failure/chemically induced , Heart Failure/drug therapy , Heart Failure/metabolismABSTRACT
BACKGROUND: Cancer cachexia is a common but severe condition that causes muscle wasting, body weight loss, and progressive functional impairment, affecting over 50% of cancer patients. Currently, there are no effective treatments that can alleviate cachexia, and hence the discovery of new therapeutics that can effectively prevent or even reverse cancer cachexia is crucial. Babao Dan (BBD) is a Traditional Chinese Medicine (TCM) formula that has been used clinically in combating various cancers, however, its therapeutic potential in alleviating cancer cachexia remains unexplored. Our current study aims to determine the anti-cachectic effects of BBD treatment in alleviating cancer cachexia, as well as determining the underlying mechanisms involved. METHODS: Mouse models of cancer cachexia were induced via implantation of CT26 colon adenocarcinoma cells, and the anti-cachectic effects and mechanisms of BBD were determined via examinations of body weight and muscle mass, as well as serum and muscle markers of cachexia and muscle atrophy. RESULTS: CT26 tumor implantation reduced in the rapid occurrence of cancer cachexia characterized by marked reductions in body weight and muscle mass, functional decrease in muscle function and accelerated deaths. BBD administration not only demonstrated robust anti-cachectic ability via preventing decreases in body weight, muscle mass, and muscle atrophy, but also markedly prolonged survival. The effects of BBD in alleviating cancer cachexia and its associated adverse effects were due to its ability in preventing the activation of IL-6/STAT3 signaling post-CT26 tumor implantation. CONCLUSION: Our findings demonstrated the robust ability of BBD in preventing cancer cachexia and alleviating the main cachexia-induced symptoms as well as prolonging survival via inhibiting activation of IL-6/STAT3 signaling pathway. Therefore, our study demonstrating the strong anti-cachectic effects of BBD in mice may provide a theoretical basis for the use of BBD as a safe and effective drug in the treatment of cancer cachexia.
Subject(s)
Adenocarcinoma , Colonic Neoplasms , Mice , Animals , Cachexia/drug therapy , Cachexia/etiology , Cachexia/metabolism , Interleukin-6 , Adenocarcinoma/drug therapy , Colonic Neoplasms/complications , Colonic Neoplasms/drug therapy , Muscular Atrophy , Signal Transduction , Body WeightABSTRACT
BACKGROUND: Myocardial infarction (MI) is the leading cause of deaths worldwide, triggering widespread and irreversible damage to the heart. Currently, there are no drugs that can reverse ischemic damage to the myocardium and hence, finding novel therapeutic agents that can limit the extent of myocardial damage following MI is crucial. Liensinine (LSN) is a naturally derived bisbenzylisoquinoline alkaloid that is known to exhibit numerous antioxidative and cardiovascular beneficial effects. However, the role of LSN in MI-induced injury and its underlying mechanisms remain unexplored. PURPOSE: Our study aims to evaluate the cardioprotective effects of LSN following MI and its underlying molecular mechanisms. METHODS: We constructed murine models of MI in order to examine the potential cardioprotective effects and mechanisms of LSN in protecting against myocardial ischemic damage both in vivo and in vitro. RESULTS: Administration with LSN strongly protected against cardiac injuries following MI by decreasing the extent of ischemic damage and improving cardiac function. Additionally, LSN was found to be a potent inhibitor of Wnt/ßcatenin signaling pathway. Hence, the beneficial effects of LSN in preventing oxidative and DNA damage following ischemia was due to its ability to inhibit aberrant activation of Wnt/ßcatenin signaling. CONCLUSIONS: Our findings reveal for the first time a novel cardioprotective role of LSN during myocardial infarction and most notably, its ability to protect cardiomyocytes against oxidative stress-induced damage via inhibiting Wnt/ß-catenin signaling. Our study therefore suggests new therapeutic potential of LSN or plants that contain the natural alkaloid LSN in ischemic heart diseases.
Subject(s)
Myocardial Infarction , Wnt Signaling Pathway , Mice , Animals , beta Catenin/metabolism , Myocardial Infarction/metabolism , Myocardium/metabolism , Myocytes, CardiacABSTRACT
Obesity rates have rapidly increased worldwide and obesity-related diseases such as hypertension and cardiovascular diseases have become leading factors for global morbidity and mortality. Currently, there are no effective treatments that can prevent or reverse obesity long-term, and hence the prevention of obesity-related adverse effects such as hypertension is critical. Qingda granule (QDG) is a condensed Traditional Chinese Medicine (TCM) formula that has been used clinically for treating hypertension, however, its effectiveness in obesity-induced hypertension and cardiac dysfunction remains explored. Mouse models of obesity via long-term feeding of high-fat high-fructose diet (HFFD) were established to examine the effect and mechanism of QDG in protecting against obesity-induced hypertension and cardiac dysfunction. C57BL/6 mice were fed with either normal diet or HFFD over a period of 16 weeks and administered with either saline or QDG for assessment of obesity-induced blood pressure and cardiac function. QDG administration demonstrated robust anti-hypertensive effects and significantly attenuated HFFD-induced elevations in blood pressures. Moreover, QDG treatment also demonstrated robust cardioprotective effects during obesity-induced hypertension by markedly improving cardiac function and preventing cardiac hypertrophy. QDG protected against obesity-induced hypertension and cardiac dysfunction was due to its ability to prevent adverse chronic activation of Akt signaling pathway during long-term feeding of HFFD. Long-term usage of QDG treatments exhibited no observable side effects and also completely prevented obesity-induced organ damage, demonstrating the feasibility and safety of prolonged use. Our findings thus elucidated the role of QDG in preventing obesity-induced hypertension and cardiac hypertrophy via inhibiting adverse activation of Akt signaling activation. Therefore, our study provides the theoretical basis for the utilization of QDG as both a safe and effective drug in the therapeutic treatment of metabolic diseases such as obesity-induced hypertension.
ABSTRACT
BACKGROUND: Myocardial infarction (MI) is the most common cause of cardiac injury, resulting in widespread and irreversible damage to the heart. The incidence of MI gives rise to the excessive production of inflammatory cytokines that further promotes myocardial dysfunction. Wnt/ß-catenin signaling pathway is adversely activated during MI and plays an important role in the modulation of the inflammatory response following tissue injury. Huoxin pill (HXP) is a Traditional Chinese Medicine formulation that has been long used in the treatment of cardiovascular diseases, however its mechanisms of cardioprotection remain unclear. METHODS: We performed murine models of MI in order to model myocardial ischemic damage and examine the effect and underlying mechanism of HXP in protecting against myocardial ischemic injury. We further constructed conditional cardiomyocyte-specific ß-catenin knockout mice and induced surgical MI in order to better understand the role of Wnt/ß-catenin signaling following myocardial infarction in the adult heart. RESULTS: HXP administration strongly protected against cardiac ischemic injury, improved cardiac function, and markedly decreased the expression of pro-inflammatory cytokines following MI. Nuclear activation of ßcatenin resulted in significantly increased nuclear translocation and activation of NF-κB. In contrast, cardiomyocyte-specific deletion of ß-catenin decreased NF-κB activation and exhibited beneficial effects following ischemic injury. Hence, HXP protected against MI-induced ischemic injury and excessive inflammatory response via inhibiting Wnt/ßcatenin signaling. CONCLUSIONS: Our study elucidated the role of HXP in protecting against ischemic myocardial injury via preventing MI-induced inflammatory response, which was mediated by its ability to inhibit adverse Wnt/ßcatenin signaling activation. Thus, our study provides the basis for the implementation of HXP as an effective therapeutic strategy in protecting against myocardial ischemic diseases.
Subject(s)
Heart Injuries , Myocardial Infarction , Animals , Cytokines/metabolism , Disease Models, Animal , Inflammation/drug therapy , Inflammation/metabolism , Mice , Myocardial Infarction/drug therapy , Myocardial Infarction/metabolism , Myocytes, Cardiac/metabolism , NF-kappa B/metabolism , Wnt Signaling Pathway , beta Catenin/metabolismABSTRACT
Myocardial infarction (MI) is one of the leading causes of death worldwide, and due to the widespread and irreversible damage caused, new therapeutic treatments are urgently needed in order to limit the degree of ischaemic damage following MI. Aberrant activation of Wnt/ß-catenin signalling pathway often occurs during cardiovascular diseases including MI, which results in excess production of reactive oxygen species (ROS) and further promotes myocardial dysfunction. Huoxin pill (HXP) is a Traditional Chinese Medicine formula that has been widely used in the treatment of coronary heart disease and angina; however, its mechanisms remain unclear. Here, we performed mouse models of MI and examined the effects and mechanisms of HXP in protecting against MI-induced ischaemic damage. Our study showed that administration with HXP robustly protected against MI-induced cardiac injuries, decreased infarct size and improved cardiac function. Moreover, HXP attenuated ischaemia-induced DNA damage occurrence in vivo and H2 O2 -induced DNA damage occurrence in vitro, via potent inhibition of adverse Wnt/ß-catenin signalling activation. Our study thus elucidated the role and mechanism of HXP in protecting against MI and oxidative stress-induced injuries and suggests new therapeutic strategies in ischaemic heart disease via inhibition of Wnt/ß-catenin signalling pathway.
Subject(s)
Drugs, Chinese Herbal/pharmacology , Myocardial Infarction/etiology , Myocardial Infarction/metabolism , Myocardial Ischemia/complications , Wnt Signaling Pathway/drug effects , Animals , Cells, Cultured , DNA Damage/drug effects , Disease Models, Animal , Drugs, Chinese Herbal/therapeutic use , Echocardiography , Heart Function Tests , Male , Medicine, Chinese Traditional , Mice , Myocardial Infarction/diagnosis , Myocardial Infarction/prevention & control , Myocardial Ischemia/diagnosis , Myocardial Ischemia/metabolism , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , Ventricular Remodeling/drug effectsABSTRACT
CONTEXT: Huoxin pill (HXP) is a commonly used TCM prescription for treatment of cardiovascular diseases. However, its mechanism in protecting against myocardial infarction (MI) remains unknown. OBJECTIVE: We performed a network pharmacology analysis to explore the bioactive ingredients, therapeutic effects, and mechanisms of HXP in protecting against MI. MATERIALS AND METHODS: HPLC was used to identify major bioactive compounds, and overlap with MI target genes were visualised. 10-Week old C57BL/6 mice were randomly assigned as: Sham-operated control, MI + Phosphate buffered saline (PBS), and MI + HXP (3 mg/mL and 9 mg/mL) treatment groups, received oral gavage administration once every two-days starting from 1-week prior to MI, and subsequently MI models were established for one-week before sacrifice. RESULTS: AKT1, VEGFA, TNF and RELA were identified as core target proteins among eighty-five candidate bioactive compounds identified in HXP with overlapping MI-related genes. HXP protection against MI was mainly via regulation of inflammatory pathways, notably TNF signalling pathway. Mouse models of MI and cardiac myoblasts demonstrated that HXP improved MI-induced injury via improving regulation of inflammatory response. DISCUSSION AND CONCLUSION: Stellasterol, deoxycholic acid, kaempferol, and quercetin are important active compounds contained in HXP with anti-inflammatory properties in the therapeutic treatment of MI. Due to the straightforward nature and effectiveness of taking oral HXP medications, our findings provide a theoretical basis for the clinical application of HXP in treating patients with angina or myocardial ischaemia. Future research into the combination of surgical procedures or medications that restore blood flow together with HXP as supportive medication would be worthwhile.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Drugs, Chinese Herbal/pharmacology , Myocardial Infarction/prevention & control , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/chemistry , Cardiotonic Agents/administration & dosage , Cardiotonic Agents/chemistry , Cardiotonic Agents/pharmacology , Chromatography, High Pressure Liquid , Disease Models, Animal , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/administration & dosage , Drugs, Chinese Herbal/chemistry , Mice , Mice, Inbred C57BL , Network Pharmacology , Signal Transduction/drug effectsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese Medicine (TCM) is being increasingly used worldwide due to its diverse efficacy and relatively low side effects. Babao Dan (BBD) is a well-known TCM formula that is currently used for the effective treatment of various cancers, however its underlying molecular mechanism remains unknown. AIM OF THE STUDY: Tumor growth and tumor recurrence are characterized by two distinct populations of cells, namely the well-differentiated cancer cells composing the majority of tumor bulk, and cancer stem cells (CSCs) involved in tumor relapse, which are both strongly associated with excessive activation of Wnt/ß-catenin signaling. Our study aims to elucidate the underlying molecular mechanisms associated with the anti-tumor proliferative effects of Babao Dan (BBD). MATERIALS AND METHODS: We used a hepatoblastoma cell line HepG2 with stem cell-like traits that harbors a constitutively active mutant of ß-catenin in order to study the anti-tumor ability of BBD via targeting Wnt/ß-catenin signaling. RESULTS: BBD robustly attenuated both the intrinsic and extrinsic activation of Wnt/ß-catenin pathway in HepG2 hepatoblastoma cells, as well as Wnt target genes. Moreover, BBD significantly inhibited both the proliferation of well-differentiated cancer cells, as well as the stem-like property of CSCs as evidenced by EpCAM, a Wnt target gene and a novel marker of cancer cell stemness. In addition, mice administered with BBD using HepG2 cell line derived xenograft model had marked reductions in tumor size and weight, as well as significantly decreased expressions of Wnt target genes and cancer cell stemness. CONCLUSION: Our findings elucidated the underlying molecular mechanisms associated with the robust anti-tumor effects of BBD via potent inhibition of Wnt/ß-catenin signaling, and implicate its use in the clinical treatment of cancers.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Drugs, Chinese Herbal/pharmacology , Hepatoblastoma/drug therapy , Liver Neoplasms/drug therapy , Animals , Cell Proliferation/drug effects , Hep G2 Cells , Hepatoblastoma/genetics , Hepatoblastoma/pathology , Humans , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Mutation , Neoplastic Stem Cells/drug effects , Wnt Signaling Pathway/drug effects , Xenograft Model Antitumor Assays , beta Catenin/geneticsABSTRACT
Vascular endothelial growth factor (VEGF) is a well-known angiogenic factor, however its ability in promoting therapeutic angiogenesis following myocardial infarction (MI) is limited. Here, we aimed to investigate whether dual treatment with insulin-like growth factor binding protein-4 (IGFBP-4), an agent that protects against early oxidative damage, can be effective in enhancing the therapeutic effect of VEGF following MI. Combined treatment with IGFBP-4 enhanced VEGF-induced angiogenesis and prevented cell damage via enhancing the expression of a key angiogenic factor angiopoietin-1. Dual treatment with the two agents synergistically decreased cardiac fibrosis markers collagen-I and collagen-III following MI. Importantly, while the protective action of IGFBP-4 occurs at an early stage of ischemic injury, the action of VEGF occurs at a later stage, at the onset angiogenesis. Our findings demonstrate that VEGF treatment alone is often not enough to protect against oxidative stress and promote post-ischemic angiogenesis, whereas the combined treatment with IGFBP4 and VEGF can utilize the dual roles of these agents to effectively protect against ischemic and oxidative injury, and promote angiogenesis. These findings provide important insights into the roles of these agents in the clinical setting, and suggest new strategies in the treatment of ischemic heart disease.
Subject(s)
Disease Models, Animal , Insulin-Like Growth Factor Binding Protein 4/metabolism , Myocardial Infarction/complications , Neovascularization, Pathologic/pathology , Vascular Endothelial Growth Factor A/metabolism , Animals , Cells, Cultured , Insulin-Like Growth Factor Binding Protein 4/genetics , Male , Mice , Mice, Inbred C57BL , Neovascularization, Pathologic/etiology , Neovascularization, Pathologic/metabolism , Vascular Endothelial Growth Factor A/geneticsABSTRACT
BACKGROUND: LRP5/6 are co-receptors in Wnt/ß-catenin pathway. Recently, we discovered multiple ß-catenin independent functions of LRP5/6 in tumor cells and in the diseased heart. Nucleoporin 37 (NUP37) is an important component of the nuclear pore complex (NPC), whose elevated expression is associated with worsened prognosis in liver cancer. Previous studies have shown that NUP37 interacted with YAP and activated YAP/TEAD signaling in liver cancer. Our preliminary findings showed a nuclear location of LRP5. We thus tested the hypothesis that LRP5 may act as a genuine regulator of YAP/TEAD signaling via modulating NUP37 in a ß-catenin-independent way. METHODS: We performed siRNA knockdown of LRP5, LRP6, or ß-catenin in liver cancer HepG2 cells to determine the effect on tumor cell proliferation. Protein expressions and interaction between LRP5 and NUP37 were determined using immunoprecipitation and western blot analyses. RESULTS: HepG2 cell proliferation was markedly inhibited by knockdown of LRP5 but not LRP6 or ß-catenin, suggesting that LRP5 has a specific, ß-catenin-independent role in inhibiting HepG2 cell proliferation. Knockdown of NUP37 by siRNA inhibited the proliferation of HepG2 cells, whereas overexpression of NUP37 reversed the decrease in cell proliferation induced by LRP5 knockdown. Immunoprecipitation assays confirmed that LRP5 bound to NUP37. Furthermore, LRP5 overexpression restored NUP37 knockdown-induced downregulation of YAP/TEAD pathway. CONCLUSIONS: LRP5 deletion attenuates cell proliferation via destabilization of NUP37, in a ß-catenin-independent manner. LRP5 therefore acts as a genuine regulator of YAP/TEAD signaling via maintaining the integrity of the NPC, and implicates a therapeutic strategy in targeting LRP5 for inhibiting liver cancer cell proliferation.
Subject(s)
Low Density Lipoprotein Receptor-Related Protein-5/deficiency , Low Density Lipoprotein Receptor-Related Protein-5/metabolism , Nuclear Pore Complex Proteins/metabolism , Adaptor Proteins, Signal Transducing/metabolism , Cell Proliferation/genetics , Hep G2 Cells , Humans , Low Density Lipoprotein Receptor-Related Protein-5/genetics , Protein Stability , RNA, Small Interfering/genetics , Signal Transduction , Transcription Factors/metabolism , YAP-Signaling ProteinsABSTRACT
PURPOSE: Lung metastasis is an important cause of breast cancer-related deaths, in which SDF-1/CXCR4 signaling pathway plays a critical role. Single transmembrane protein LRP6 is viewed as an oncogene via activating the Wnt/ß-catenin signaling pathway. Our work aims to investigate the relationship between SDF-1/CXCR4 and LRP6 in breast cancer lung metastasis. EXPERIMENTAL DESIGN: We examined the expressions and functions of SDF-1/CXCR4 and LRP6 as well as their relationship in breast cancer in vitro and in vivo. RESULTS: LRP6 ectodomain (LRP6N) directly bound to CXCR4 and competitively prevented SDF-1 binding to CXCR4. LRP6N prevented SDF-1/CXCR4-induced metastasis to lung and prolonged survival in mice bearing breast tumors, whereas LRP6 knockdown activated SDF-1/CXCR4 signal transduction and promoted lung metastasis and tumor death. Furthermore, patients with breast cancer with high CXCR4 expression had poor prognosis, which was exacerbated by low LRP6 expression but improved by high LRP6 expression. Interestingly, a secreted LRP6N was found in the serum of mice and humans, which was downregulated by the onset of cancer metastasis in both mice bearing breast cancer as well as in patients with breast cancer. CONCLUSIONS: LRP6N might be a promising diagnostic marker for the early detection of breast cancer metastasis as well as an inhibitor of SDF-1/CXCR4-induced breast cancer metastasis. LRP6N also provides an interesting link between Wnt signaling and SDF-1/CXCR4 signaling, the two key pathways involved in cancer development.
Subject(s)
Biomarkers, Tumor/metabolism , Breast Neoplasms/prevention & control , Chemokine CXCL12/adverse effects , Chemokine CXCL12/metabolism , Low Density Lipoprotein Receptor-Related Protein-6/metabolism , Lung Neoplasms/prevention & control , Receptors, CXCR4/metabolism , Adolescent , Adult , Aged , Animals , Breast Neoplasms/etiology , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Cell Line, Tumor , Cell Movement , Female , Humans , Lung Neoplasms/metabolism , Lung Neoplasms/secondary , Mice , Mice, Inbred BALB C , Mice, Nude , Middle Aged , Prognosis , Receptors, CXCR4/administration & dosage , Survival Rate , Wnt Signaling Pathway , Xenograft Model Antitumor Assays , Young AdultABSTRACT
The protective role of alkaloids from Nelumbinis Plumula (AFNP) on the aorta during hypertension is not yet fully understood. We hypothesize that AFNP exerts protective effects against Ang II-induced hypertension by mediating RhoA/ROCK pathway and phenotypic switching during hypertension. In the present study, we evaluated the effect of AFNP on angiotensin II (Ang II)-induced actin cytoskeleton reorganization and aorta remodeling, as well as the involvement of RhoA/Rho-associated coiled kinase (ROCK) pathway in protecting against hypertension. We used rat aortic tissues to investigate the vasodilatation effect of AFNP on Ang II-induced constriction. AFNP was shown to significantly relax the endothelium-intact arteries induced by Ang II. We further investigated the vasodilation effect of AFNP in endothelium denuded arteries, which showed that the action of AFNP was endothelial independent. Male SHR rats were treated with saline or AFNP and morphological changes were examined following 8 weeks. AFNP treatment normalized the effects of hypertension in SHRs. HE staining showed that AFNP treatment improved the tunica media and wall thickness and ratio of MT/LD and MA/LA. Western blotting showed that AFNP treatment markedly decreased the Ang II-induced expression of collagen I and increased α-SMA in aorta. Furthermore, MTT assay showed that AFNP inhibited the proliferation of Ang II treated VSMCs in a concentration-dependent manner. AFNP treatment also ameliorated F-actin cytoskeleton remodeling in Ang II treated VSMCs, as visualized under fluorescence microscopy. Western blot analysis showed that RhoA transposition and ROCK activation and phosphorylation of MYPT1 was increased following Ang II treatment but were inhibited by AFNP treatment, showing that the cardio-protective effect of AFNP is likely mediated by the RhoA/ROCK signaling pathway. The anti-hypertension and aortic protection effects of AFNP are due to non-endothelial dependent inhibition of the VSMC cytoskeleton remodeling and regulation of RhoA/ROCK pathway.
Subject(s)
Alkaloids/pharmacology , Aorta, Thoracic/drug effects , Nelumbo , Vascular Remodeling/drug effects , rho GTP-Binding Proteins/physiology , rho-Associated Kinases/physiology , Alkaloids/isolation & purification , Animals , Aorta, Thoracic/physiology , Cell Survival/drug effects , Cell Survival/physiology , Dose-Response Relationship, Drug , Male , Rats , Rats, Inbred SHR , Rats, Inbred WKY , Rats, Sprague-Dawley , Seeds , Signal Transduction/drug effects , Signal Transduction/physiology , Vascular Remodeling/physiologyABSTRACT
BACKGROUND: The Midwife-led maternity services have been implemented in China in response to the high rates of primiparous women and Caesarean Sections (CS) which may be related to China's one-child policy. However, few studies in China have been reported on the effectiveness of Midwife-led Care at Delivery (MCD) and the Continuity of Midwife-led Care (CMC) on postpartum wellbeing and other clinical outcomes. Therefore, evidence-based clinical validation is needed to develop an optimal maternity service for childbearing women in China. METHODS: A concurrent cohort study design was conducted with 1730 pregnant women recruited from 9 hospitals in Shanghai. Among the 1730 participants at baseline, 1568 participants completed the follow-up questionnaire, with a follow-up rate of 90.6%. RESULTS: Compared with the routine Obstetrician-led Maternity Care (OMC), Midwife-led Care at Delivery (MCD) was associated with CS rate (OR were 0.16; 95%CI: 0.11 to 0.25) and a higher total score of postpartum wellbeing (ßwere 2.70; 95%CI: 0.70 to 4.70) when adjusting for the baseline differences and other confounders during delivery or postpartum period. Moreover, continuity of Midwife-led Care (CMC) was associated with CS rate (OR were 0.30; 95%CI: 0.23 to 0.41), as well as increased rate of breastfeeding within the first 24 h (OR were 2.49; 95% CI: 1.47 to 4.23), higher postpartum satisfaction (ß = 4.52; 95% CI: 1.60 to 12.68), lower anxiety (ßwere 0.66; 95% CI: 0.16 to 1.17), increased self-control (ßwere 0.39; 95% CI: 0.02 to 0.76) and a higher total score of postpartum wellbeing (ßwere 3.14; 95% CI: 1.54 to 4.75). CONCLUSION: CMC is the optimal service for low-risk primiparous women under China's one-child policy, and is worthwhile for a general implementation across China.
Subject(s)
Delivery of Health Care/methods , Maternal Health Services/organization & administration , Mental Health , Midwifery , Postpartum Period , Adult , Anxiety , Breast Feeding/statistics & numerical data , Cesarean Section/statistics & numerical data , China , Family Planning Policy , Female , Humans , Obstetrics , Parity , Patient Satisfaction/statistics & numerical data , Perinatal Care , Pregnancy , Self-Control , Young AdultABSTRACT
Baicalin is one of the active ingredients extracted from the dry root of Scutellaria baicalensis Georgi, which has been reported to be effective in preventing myocardial ischemia reperfusion injury. However, the mechanisms underlying its cardioprotective activities remain to be elucidated. In the present study, H2O2treated cardiomyocyte H9c2 cell line served as an in vitro model of oxidationdamaged cardiomyocytes to evaluate the effects of baicalin on the cardiac injury, and to investigate the underlying molecular mechanism. The results of the TOPFlash/Renilla reporter gene assay indicated that baicalin significantly suppressed the activation of protooncogene Wnt1 (Wnt)/ßcatenin in 293 cells, in a dose dependent manner. In addition, baicalin significantly inhibited H2O2induced loss of H9c2 cell viability in MTT assay. Furthermore, western blotting analysis demonstrated that baicalin markedly attenuated H2O2induced cell apoptosis, as demonstrated by the downregulation of cleaved caspase3 and the increase in the apoptosis regulator Bcl2 (Bcl2)/apoptosis regulator BAX (Bax) ratio following baicalin treatment in H2O2treated H9c2 cells. Furthermore, baicalin markedly decreased the expression of ßcatenin and downstream Axin2 and myc protooncogene protein in H2O2treated H9c2 cells. Knockdown of ßcatenin expression inhibited H2O2induced cell apoptosis. Finally, LiCl (a ßcatenin stabilizer) induced apoptosis of H9c2 cells by upregulating the expression of ßcatenin, which was significantly neutralized by the treatment with baicalin. Taken together, it is hypothesized that baicalin exerts cardioprotective effects via suppression of the Wnt/ßcatenin signaling pathway.
Subject(s)
Antioxidants/pharmacology , Flavonoids/pharmacology , Hydrogen Peroxide/pharmacology , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/metabolism , Wnt Signaling Pathway/drug effects , Apoptosis/drug effects , Cell Survival/drug effects , Cells, Cultured , Gene Expression , Genes, Reporter , Humans , Oxidative Stress/drug effects , beta Catenin/genetics , beta Catenin/metabolismABSTRACT
BACKGROUND: Myocardial infarction is one of the leading causes of morbidity and mortality worldwide, triggering irreversible myocardial cell damage and heart failure. The role of low-density lipoprotein receptor-related proteins 5 and 6 (LRP5/6) as coreceptors of the Wnt/ß-catenin pathway in the adult heart remain unknown. Insulin-like growth factor binding protein 4 and dickkopf-related protein 1 (Dkk1) are 2 secreted LRP5/6 binding proteins that play a crucial role in heart development through preventing Wnt/ß-catenin pathway activation. However, their roles in the adult heart remain unexplored. METHODS: To understand the role of LRP5/6 and ß-catenin in the adult heart, we constructed conditional cardiomyocyte-specific LRP5/6 and ß-catenin knockout mice and induced surgical myocardial infarction. We also directly injected recombinant proteins of insulin-like growth factor binding protein 4 and Dkk1 into the heart immediately following myocardial infarction to further examine the mechanisms through which these proteins regulate LRP5/6 and ß-catenin. RESULTS: Deletion of LRP5/6 promoted cardiac ischemic insults. Conversely, deficiency of ß-catenin, a downstream target of LRP5/6, was beneficial in ischemic injury. It is interesting to note that although both insulin-like growth factor binding protein 4 and Dkk1 are secreted Wnt/ß-catenin pathway inhibitors, insulin-like growth factor binding protein 4 protected the ischemic heart by inhibiting ß-catenin, whereas Dkk1 enhanced the injury response mainly through inducing LRP5/6 endocytosis and degradation. CONCLUSIONS: Our findings reveal previously unidentified dual roles of LRP5/6 involved in the cardiomyocyte response to ischemic injury. These findings suggest new therapeutic strategies in ischemic heart disease by fine-tuning LRP5/6 and ß-catenin signaling within the Wnt/ß-catenin pathway.
Subject(s)
Insulin-Like Growth Factor Binding Protein 4/pharmacology , Intercellular Signaling Peptides and Proteins/pharmacology , Low Density Lipoprotein Receptor-Related Protein-5/metabolism , Low Density Lipoprotein Receptor-Related Protein-6/metabolism , Myocardial Ischemia/pathology , Wnt Signaling Pathway/drug effects , beta Catenin/metabolism , Animals , DNA Damage/drug effects , Disease Models, Animal , Down-Regulation/drug effects , Histones/metabolism , Hydrogen Peroxide/toxicity , Insulin-Like Growth Factor Binding Protein 4/genetics , Insulin-Like Growth Factor Binding Protein 4/therapeutic use , Intercellular Signaling Peptides and Proteins/genetics , Intercellular Signaling Peptides and Proteins/therapeutic use , Low Density Lipoprotein Receptor-Related Protein-5/antagonists & inhibitors , Low Density Lipoprotein Receptor-Related Protein-5/genetics , Low Density Lipoprotein Receptor-Related Protein-6/antagonists & inhibitors , Low Density Lipoprotein Receptor-Related Protein-6/genetics , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Myocardial Ischemia/metabolism , Myocardial Ischemia/prevention & control , Myocytes, Cardiac/cytology , Myocytes, Cardiac/metabolism , RNA Interference , RNA, Small Interfering/metabolism , Recombinant Proteins/biosynthesis , Recombinant Proteins/pharmacology , Recombinant Proteins/therapeutic use , Wnt Proteins/antagonists & inhibitors , Wnt Proteins/metabolism , beta Catenin/antagonists & inhibitors , beta Catenin/geneticsABSTRACT
Nuclear respiratory factor 1 (NRF-1) has important roles in the regulation of several key metabolic genes required for cellular growth and respiration. A previous study by our group indicated that NRF1 is involved in mitochondrial dysfunction induced by the environmental pollutant benzo[a]pyrene in the 16HBE human bronchial epithelial cell line. In the present study, it was observed that its genotoxic metabolite, antibenzopyrene7,8diol9,10epoxide (BPDE), triggered cell cycle arrest in Sphase in 16HBE cells by activating ataxia-telangiectasia (ATM)/checkpoint kinase (Chk)2 and ATM and Rad3 related (ATR)/Chk1 signaling pathways. NRF1 expression was suppressed by BPDE after treatment for 6 h. Flow cytometric analysis revealed that NRF1 overexpression attenuated cell cycle arrest in Sphase induced by BPDE. In line with this result, DNAdamage checkpoints were activated following NRF1 overexpression, as demonstrated by increased phosphorylation of ATM, Chk2 and γH2AX, but not ATR and Chk1, according to western blot analysis. It was therefore indicated that NRF1 overexpression attenuated BPDEinduced Sphase arrest via the ATM/Chk2 signaling pathway.
Subject(s)
Bronchi/metabolism , Dihydroxydihydrobenzopyrenes/toxicity , Epithelial Cells/metabolism , Gene Expression Regulation/drug effects , Nuclear Respiratory Factor 1/biosynthesis , Respiratory Mucosa/metabolism , S Phase Cell Cycle Checkpoints/drug effects , Ataxia Telangiectasia Mutated Proteins/genetics , Ataxia Telangiectasia Mutated Proteins/metabolism , Bronchi/pathology , Checkpoint Kinase 2/genetics , Checkpoint Kinase 2/metabolism , Epithelial Cells/pathology , Gene Expression Regulation/genetics , Humans , Nuclear Respiratory Factor 1/genetics , Respiratory Mucosa/pathology , S Phase Cell Cycle Checkpoints/genetics , Signal Transduction/drug effects , Signal Transduction/geneticsABSTRACT
AIM: The aim of this study was to examine the effects of home and educational environments on children's motor performance in China. METHOD: We conducted a cross-sectional study of 4001 preschool children selected from 160 classes. The children's motor performance was assessed using the Movement Assessment Battery for Children, 2nd edition (MABC-2). Home and educational environments were evaluated using validated checklists. The effects of home and educational environments on motor performance were analysed using mixed and multilevel logistic regression models. RESULTS: The results showed that one score increase in the outside space of the family home was positively associated with the increase in total test score (0.104) subtest score of aiming and catching (0.037), and balance (0.034) of the MABC-2, after adjusting for potential confounders (each p<0.05). Possession of motor toys at home and parental rearing behaviours were also related to total test score, manual dexterity, and balance (ß=0.022-0.104, each p<0.05). Space and furnishings, activity, and interaction in the classroom had a significant positive association with total test score (ß=0.069-0.201), and with subtest scores of manual dexterity, aiming and catching, and balance respectively (ß=0.115-0.206). Space and furnishings of classrooms and possession of toys in the household were protective factors for 'at risk' or significant poor performance (odds ratio 0.942-0.973, each p<0.05). INTERPRETATION: A permissive and accepting family and educational environment made a positive contribution to children's motor performance. Access to sufficient space and furnishings within the classroom, as well as toys in the family, were protective factors for poor motor performance. Future assistance is needed to support an advantageous environment in early childhood programmes in China.