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OBJECTIVE: This study investigated the potential of the slow-developing blastocysts using preimplantation genetic testing-aneuploidy (PGT-A) in patients undergoing frozen-thawed embryo transfer, stratified by age. STUDY DESIGN: A retrospective analysis was performed including a total of 743, the first frozen embryo transfer (FET) cycle with single embryo transfer (SET), who underwent treatment between January 2020 and July 2023 in a single fertility center, XXXX Fertility Center. A total of 743 cycles, in which we performed intracellular sperm injection and freeze all strategy, from 743 patients were included. The patient group was divided into 4 groups as follows: Group 1 (G1), 208 FET on day 5; Group 2 (G2), 177 FET with PGT-A on day 5; Group 3 (G3), 220 FET on day 6; Group 4 (G4), 138 FET with PGT-A on day 6. We also divided into two groups-under 35 years of age and over 35 years of age-and performed the analysis separately for each group. RESULTS: In under 35 years of age groups, there were no significant differences in clinical pregnancy and miscarriage rates in G1 and G2 (67.2% vs 63.8%, NS). Also, G4 had a higher clinical pregnancy rate than G3, but it was not significant (51.8% vs 54.7%, NS). In 35 years or older group, G2 had higher pregnancy rates than G1 and lower miscarriage rates (CPR: 43.3% vs 67.7%, P = 0.001, MR: 22.5% vs 3.4%, P = 0.001). In addition, G4 had a higher pregnancy rate than G3 and lower miscarriage rate (CPR: 31.8% vs. 46.9%, P = 0.003, MR: 22.9% vs 2.2%, P = 0.023). CONCLUSIONS: In ≥35 years group, PGT-A on day 5 and day 6 showed a high pregnancy rate and a low miscarriage rate. Therefore, using PGT-A seems advantageous for patients of an advanced maternal age.
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AIM: To evaluate the relationship between AMH and ovarian response to controlled ovarian hyperstimulation in women with PCOM and PCOS. METHODS: A retrospective study was conducted on 559 patients who underwent the IVF-ET cycle between January 2018 and December 2022 at Gangnam Cha Hospital. Patients were divided into 3 groups matched for age and BMI: the PCOS group (n = 54), based on the new 2023 PCOS guideline; the PCOM group (n = 53); and the control group (n = 452) with normal ovaries. Serum AMH levels were converted to multiples of the median (MoM) for each corresponding age. The ovarian sensitivity index (OSI) was calculated as the number of retrieved oocytes divided by the total dose of recombinant FSH administered (per 1000 IU). RESULTS: There were significant differences in AMH-MoM value among women with PCOS [2.7 ± 1.3 (95% CI 2.3-3.0)], those with PCOM [2.0 ± 1.0 (95% CI 1.7-2.3)], and controls [0.8 ± 0.7 (95% CI 0.8-0.9)] (p < 0.001). The abortion rates in the normoovulatory, PCOM, and PCOS groups were 18.2%, 21.1%, and 25.0%, respectively. OSI and live birth rate were positively correlated with the AMH-MoM value in normoovulatory women (r = 0.389, p < 0.05, r = 0.122, p < 0.05), while no such correlation was observed in women with PCOM and PCOS. CONCLUSIONS: Ovarian response and live birth rate are possibly correlated with the AMH-MoM value in normoovulatory women, but not in women with PCOM and PCOS.
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OBJECTIVES: This study aimed to evaluate the clinical efficacy of fertility-preserving therapy through in vitro fertilization (IVF) procedures in women who were pathologically diagnosed with endometrial hyperplasia or carcinoma. DESIGN: A retrospective cohort study on fertility-preserving therapy was conducted. Participants/Materials, Setting: A total of 82 women were enrolled who had simple endometrial hyperplasia (SH), complex hyperplasia (CH), complex atypical hyperplasia (CAH), and endometrioid endometrial carcinoma stage IA (EC IA) and underwent IVF at Gangnam CHA fertility center between January 2008 and December 2020. METHODS: The primary endpoints were oncologic outcomes and subsequent reproductive outcomes of patients who underwent fertility-preserving treatments analyzed by χ2 test or Fisher's exact test. RESULTS: Of the 82 patients, 33 had a cumulative clinical pregnancy (40.2%), and 25 had a cumulative live birth (30.5%) through IVF procedures following pathologic confirmation of complete remission or non-progressive status. The cumulative clinical pregnancy rates and live birth rates for SH were 50.0% and 30.0%, for CH were 37.8% and 28.9%, for CAH were 25.0% and 25.0%, and for EC were 38.5% and 38.5%, respectively. There were no significant differences in cumulative clinical pregnancy rates or live birth rates when comparing the four groups. There was a difference in endometrial thickness between medroxyprogesterone acetate (MPA) treatment group and intrauterine device (IUD) group (p = 0.036); however, there were no significant differences in clinical pregnancy rates among MPA, IUD, and MPA+IUD groups. LIMITATIONS: Because of the retrospective nature of the study, many factors relevant to the treatment decision were not strictly controlled. CONCLUSIONS: All endometrial hyperplasia and carcinoma groups had competent cumulative live birth rates by IVF procedures. There may be differences in endometrial thickness depending on the treatment methods, but this does not affect clinical pregnancy rates. Therefore, the fertility-preserving treatment for endometrial hyperplasia and carcinoma is a safe and feasible method that results in good IVF outcomes.
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Selective fascicular involvement of the median nerve trunk above the elbow leading to anterior interosseous nerve (AIN) syndrome is a rare form of peripheral neuropathy. This condition has recently garnered increased attention within the medical community owing to advancements in imaging techniques and a growing number of reported cases. In this article, we explore the topographical anatomy of the median nerve trunk and the clinical features associated with AIN palsy. Our focus extends to unique manifestations captured through MRI and ultrasonography (US) studies, highlighting noteworthy findings, such as nerve fascicle swelling, incomplete constrictions, hourglass-like constrictions, and torsions, particularly in the posterior/posteromedial region of the median nerve. Surgical observations have further enhanced the understanding of this complex neuropathic condition. High-resolution MRI not only reveals denervation changes in the AIN and median nerve territories but also illuminates these alterations without the presence of compressing structures. The pivotal roles of high-resolution MRI and US in diagnosing this condition and guiding the formulation of an optimal treatment strategy are emphasized.
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Imageamento por Ressonância Magnética , Nervo Mediano , Ultrassonografia , Humanos , Imageamento por Ressonância Magnética/métodos , Nervo Mediano/diagnóstico por imagem , Ultrassonografia/métodos , Braço/inervação , Braço/diagnóstico por imagem , Neuropatia Mediana/diagnóstico por imagem , SíndromeRESUMO
In this study, ferulic acid was investigated for its potential in suppressing TNF-α-treated inflammation and insulin resistance in adipocytes. Ferulic acid suppressed TNF-α, IL-6, IL-1ß, and MCP-1. TNF-α increased p-JNK and ERK1/2, but treatment with ferulic acid (1, 10, and 50 µM) decreased p-JNK and ERK1/2. TNF-α induced the activation of IKK, IκBα, and NF-κB p65 compared to the control, but ferulic acid inhibited the activation of IKK, IκBα, and NF-κB p65. Following treatment with TNF-α, pIRS-1ser307 increased and pIRS-1tyr612 decreased compared to the control. Conversely, as a result of treatment with 1, 10, and 50 µM ferulic acid, pIRS-1ser307 was suppressed, and pIRS-1tyr612 was increased. Therefore, ferulic acid reduced inflammatory cytokine secretion by regulating JNK, ERK, and NF-κB and improved insulin resistance by suppressing pIRS-1ser. These findings indicate that ferulic acid can improve inflammation and insulin resistance in adipocytes.
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Ácidos Cumáricos , Resistência à Insulina , NF-kappa B , Camundongos , Animais , Inibidor de NF-kappaB alfa , Fator de Necrose Tumoral alfa , Células 3T3-L1 , Inflamação/tratamento farmacológico , AdipócitosRESUMO
Oxidative stress is a major cause of hepatic insulin resistance. This study investigated whether (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone), a homoisoflavonoid compound isolated from Portulaca oleracea L., alleviates insulin resistance and inhibits gluconeogenesis by reducing palmitate (PA)-induced reactive oxygen species (ROS)/c-Jun NH2-terminal kinase (JNK) activation in HepG2 cells. PA treatment (0.5 mM) for 16 h resulted in the highest production of ROS and induced insulin resistance in HepG2 cells. HM-chromanone, like N-acetyl-1-cysteine, significantly decreased PA-induced ROS production in the cells. HM-chromanone also significantly inhibited PA-induced JNK activation, showing a significant reduction in tumor necrosis factor and interleukin expression levels. Thus, HM-chromanone decreased the phosphorylation of Ser307 in insulin receptor substrate 1, while increasing phosphorylation of serine-threonine kinase (AKT), thereby restoring the insulin signaling pathway impaired by PA. HM-chromanone also significantly increased the phosphorylation of forkhead box protein O, thereby inhibiting the expression of gluconeogenic enzymes and reducing glucose production in PA-treated HepG2 cells. HM-chromanone also increased glycogen synthesis by phosphorylating glycogen synthase kinase-3ß. Therefore, HM-chromanone may alleviate insulin resistance and inhibit gluconeogenesis by regulating PA-induced ROS/JNK activation in HepG2 cells.
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This study investigated whether scopoletin could protect INS-1 pancreatic ß cells from apoptosis and oxidative stress caused by high glucose. Cells were pretreated with glucose (5.5 or 30 mM) and then treated with 0, 5, 10, 25, or 50 µM Scopoletin. Cell viability and insulin secretion were measured in addition to ROS, TBARS, NO and antioxidant enzymes. Western blot analysis and flow cytometric assessment of apoptosis were also carried out. High glucose of 30 mM caused glucotoxicity and cell death in INS-1 pancreatic ß cells. However, 5, 10, 25 or 50 µM scopoletin increased the level of cell viability as concentrations increased. The levels of ROS, TBARS, and NO increased by high glucose were significantly decreased after scopoletin treatment. Scopoletin also raised antioxidant enzyme activities up against oxidative stress produced by high glucose. These effects influenced the apoptosis pathway, raising levels of anti-apoptotic protein, Bcl-2, and reducing levels of pro-apoptotic proteins, including JNK, Bax, cytochrome C, and caspase 9. Annexin V/propidium staining indicated that scopoletin significantly lowered high glucose-produced apoptosis. These results indicate that scopoletin can protect INS-1 pancreatic ß cells from glucotoxicity caused by high glucose and have potential as a pharmaceutical material to protect the pancreatic ß cells.
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Células Secretoras de Insulina , Células Secretoras de Insulina/metabolismo , Escopoletina/farmacologia , Escopoletina/metabolismo , Antioxidantes/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Apoptose , Estresse Oxidativo , Glucose/toxicidade , Glucose/metabolismo , Insulina/metabolismoRESUMO
Type 2 diabetes is a disease characterized by hyperglycemia and is a growing health problem worldwide. Since many known diabetes drugs are side effects, it is necessary to develop natural substances with guaranteed safety. HM-chromanone isolated from Portulaca oleracea L. is a homoisoflavonoid compound. We investigated the effects of HM-chromanone on hyperglycemia and its mechanism in C57BL/6J ob/ob mice. C57BL/6J-Jms Slc mice were used as the control group, and C57BL/6J-ob/ob mice were divided into three groups: ob/ob (control), metformin (Met; positive control), and HM-chromanone (HMC). Fasting blood glucose was lower in the HMC group than those in the ob/ob group. Insulin resistance was improved by reducing HbA1c, plasma insulin, and HOMA-IR levels in the HMC group. HMC administration decreased the phosphorylation of IRS-1ser307 and increased the phosphorylation of IRS-1tyr612, PI3K, phosphorylation of AKTser473, and PM-GLUT4 in the skeletal muscles of ob/ob mice, indicating improved insulin signaling. HMC administration also increased the phosphorylation of FOXO1 in the liver of ob/ob mice. This inhibited PEPCK and G6pase involved in gluconeogenesis and regulated phosphorylation of glycogen synthase kinase 3ß and glycogen synthase involved in glycogen synthesis. In conclusion, HM-chromanone ameliorates hyperglycemia by PI3K/AKT and improves the FOXO1 in ob/ob mice.
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Diabetes Mellitus Tipo 2 , Hiperglicemia , Insulinas , Camundongos , Animais , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Camundongos Endogâmicos C57BL , Diabetes Mellitus Tipo 2/tratamento farmacológico , Camundongos Endogâmicos , Hiperglicemia/tratamento farmacológicoRESUMO
We examined the effects of HM-chromanone (HMC) on alleviating hyperglycemia and protecting pancreatic ß-cells from streptozotocin (STZ)-induced damage in C57BL/6J mice. HMC was administered to STZ-induced diabetic mice at 10 or 30 mg/kg, for 14 days. Thereafter, changes in fasting blood glucose levels, insulin-secretion, histopathological examination of pancreas islet cell and apoptotic protein levels, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were determined. The results revealed that HMC dose-dependently improved blood glucose concentrations and alleviated pancreatic islet cells damage. In diabetic mice, degeneration of the islet cells was observed wherein they appeared shrunken, with hyaline deterioration, nuclear dissolution, and condensation. However, morphology of the islet cell was restored, and nuclei were visibly rounded in the HMC (30 mg/kg)-administered diabetic mice. In addition, ß-cell numbers were markedly increased in HMC mice compared to STZ-induced diabetic mice, and the number of cells stained with glucagon was decreased. HMC markedly decreased the expression of proapoptotic proteins and increased antiapoptotic proteins, and the number of apoptotic cells detected by TUNEL was elevated. HMC decreased expression of interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α in diabetic mice. Moreover, HMC increased antioxidant-enzymes activity, and decreased reactive oxygen species generation. In conclusion, the results demonstrate the potential of HMC to alleviate hyperglycemia by protecting the pancreatic ß-cells in diabetic mice.
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Diabetes Mellitus Experimental , Hiperglicemia , Células Secretoras de Insulina , Ilhotas Pancreáticas , Camundongos , Animais , Estreptozocina/efeitos adversos , Insulina , Glicemia/metabolismo , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Camundongos Endogâmicos C57BL , Ilhotas Pancreáticas/metabolismo , Hiperglicemia/tratamento farmacológico , Hiperglicemia/metabolismo , Células Secretoras de Insulina/metabolismo , Antioxidantes/farmacologia , Antioxidantes/metabolismoRESUMO
This study aimed to identify the effect of (E)-5-hydroxy-7-methoxy-3-(2'-hydroxybenzyl)-4-chromanone (HM-chromanone), isolated from Portulaca oleracea L., on tyrosine phosphatase 1B (PTP1B) and glucose production in insulin-resistant HepG2 cells. The results revealed that HM-chromanone significantly decreases PTP1B expression and glucose production in insulin-resistant HepG2 cells. Furthermore, a molecular docking stimulation showed HM-chromanone inhibits PTP1B by binding to its active site. Additionally, HM-chromanone was found to significantly modulate insulin receptor substrate-1 (IRS1) by decreasing phosphorylated serine 307 and increasing phosphorylated tyrosine 612 and activating phosphatidylinositol 3-kinase (PI3K) in insulin-resistant HepG2 cells. Furthermore, HM-chromanone augmented the phosphorylation of Akt and forkhead box protein O1 in insulin-resistant HepG2 cells in a dose-dependent manner at the concentrations of 15-60 µM. Additionally, it significantly reduced the expression of glucose 6-phosphatase and phosphoenolpyruvate carboxykinase, which are main enzymes included in hepatic gluconeogenesis. Consequently, HM-chromanone was confirmed to significantly decrease glucose production and increase glucose uptake in insulin-resistant HepG2 cells.
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Resistência à Insulina , Portulaca , Humanos , Insulina/metabolismo , Glucose/metabolismo , Células Hep G2 , Portulaca/química , Fosfatidilinositol 3-Quinases/metabolismo , Transdução de Sinais , Proteína Tirosina Fosfatase não Receptora Tipo 1 , Simulação de Acoplamento Molecular , Proteínas Proto-Oncogênicas c-akt/metabolismo , Resistência à Insulina/fisiologia , Estrutura Molecular , TirosinaRESUMO
Increased free fatty acid levels in the blood are common in obesity and cause insulin resistance associated with type 2 diabetes in the muscles. Previous studies have confirmed the antidiabetic and anti-obesity potential of (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone). However, it is unknown how HM-chromanone alleviates obesity-related insulin resistance in L6 skeletal muscle cells. Palmitate induced insulin resistance and reduced glucose uptake, whereas HM-chromanone significantly increased glucose uptake. In palmitate-treated L6 skeletal muscle cells, HM-chromanone stimulated liver kinase B1 (LKB1) and 5'-adenosine monophosphate-activated protein kinase (AMPK) phosphorylation. The AMPK inhibitor compound C, and the LKB1 inhibitor radicicol blocked the effects of HM-chromanone. Furthermore, HM-chromanone significantly inhibited mammalian target of rapamycin (mTOR) and ribosomal protein S6 kinase 1 (S6K1) activation, but there was no change in protein kinase C θ (PKC θ) expression. When pAMPK was inhibited with compound C, the effect of HM-chromanone on the inhibition of mTOR and S6K1 was significantly diminished. This indicates that HM-chromanone inhibits mTOR and S6K1 activation through pAMPK activation. Inhibition of mTOR and S6K1 by HM-chromanone significantly reduced IRS-1Ser307 and IRS-1Ser632 phosphorylation, leading to insulin resistance. This resulted in an increase in PM-GLUT4 (glucose transporter 4) expression, thereby stimulating glucose uptake in insulin-resistant muscle cells. HM-chromanone can improve palmitate-induced insulin resistance by inhibiting mTOR and S6K1 through activation of the AMPK pathway in L6 skeletal muscle cells. These results show the therapeutic potential of HM-chromanone for improving insulin resistance in type 2 diabetes.
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This study investigated whether (E)-5-hydroxy-7-methoxy-3-(2'-hydroxybenzyl)-4-chromanone (HM-chromanone) could counteract the high glucose level-induced blockade of insulin signaling in human HepG2 cells. Cells were pre-incubated with glucose (5.5 or 33 mM) and then incubated with a medium containing various concentrations of HM-chromanone. Assays for glucose uptake, glycogen synthesis, and glucose production were performed. Western blotting helped elucidate the underlying molecular mechanisms. High glucose concentration (33 mM) significantly increased p-IRS-1ser307 levels and decreased p-Akt levels. However, HM-chromanone significantly decreased p-IRS-1ser307 levels while increasing p-IRS-1tyr612 and Akt levels, which restored insulin signaling disturbed by high glucose concentration. HM-chromanone significantly increased p-AMPK levels, which were reduced by high glucose in HepG2 cells. Knockdown of AMPK using siRNA increased p-IRS-1ser307 and decreased p-Akt levels, even after treatment with HM-chromanone in high glucose concentration-treated cells. HM-chromanone stimulated glycogen synthesis by increasing p-GSK3ßser9 and decreasing p-GSser641 levels in HepG2 cells under high glucose concentration; this effect was blocked by AMPK siRNA. HM-chromanone significantly decreased PEPCK, G6Pase, and hepatic glucose production, which were also blocked by AMPK siRNA. These results suggest that HM-chromanone could reverse insulin signaling blockade (induced by high glucose levels) through the activation of AMPK and stimulation of glucose uptake and glycogen synthesis in HepG2 cells.
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Resistência à Insulina , Insulina , Humanos , Insulina/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células Hep G2 , Proteínas Quinases Ativadas por AMP/metabolismo , RNA Interferente Pequeno , Fosforilação , Glucose , GlicogênioRESUMO
This study investigated the effect of (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone) on palmitate-induced insulin resistance and elucidated the underlying mechanism in L6 skeletal muscle cells. Glucose uptake was markedly decreased due to palmitate-induced insulin resistance in these cells; however, 10, 25, and 50 µM HM-chromanone remarkably improved glucose uptake in a concentration-dependent manner. HM-chromanone treatment downregulated protein tyrosine phosphatase 1B (PTP1B) and phosphorylation of c-Jun N-terminal kinase (JNK) and inhibitor of nuclear factor kappa-B kinase subunit beta (IKKß), which increased because of palmitate mediating the insulin-resistance status in cells. HM-chromanone promoted insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation and suppressed palmitate-induced phosphorylation of IRS-1 serine. This activated phosphoinositide 3-kinase (PI3K) and stimulated protein kinase B (AKT) phosphorylation. Phosphorylated AKT promoted the translocation of Glucose transporter type 4 to the plasma membrane and significantly enhanced glucose uptake into muscle cells. Additionally, HM-chromanone increased glycogen synthesis through phosphorylating glycogen synthase kinase 3 alpha/beta (GSK3 α/ß) via AKT. Consequently, HM-chromanone may improve insulin resistance by downregulating the phosphorylation of IRS-1 serine through inhibition of negative regulators of insulin signaling and inflammation-activated protein kinases in L6 skeletal muscle cells.
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Resistência à Insulina , Portulaca , Glucose/metabolismo , Transportador de Glucose Tipo 4/metabolismo , Glicogênio/metabolismo , Quinase 3 da Glicogênio Sintase/metabolismo , Humanos , Quinase I-kappa B/metabolismo , Insulina/metabolismo , Proteínas Substratos do Receptor de Insulina/metabolismo , Resistência à Insulina/fisiologia , Isoflavonas , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Fibras Musculares Esqueléticas , Músculo Esquelético/metabolismo , Palmitatos/metabolismo , Palmitatos/farmacologia , Fosfatidilinositol 3-Quinase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Portulaca/química , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serina/metabolismo , Tirosina/metabolismoRESUMO
While a number of coding genes have explained the anticancer activity of ginsenoside Rh2, little is known about noncoding RNAs. This study was performed to elucidate the regulatory activity of long noncoding RNA (lncRNA) CFAP20DC-AS1, which is known to be downregulated by Rh2. MiR-3614-3p, which potentially binds CFAP20DC-AS1, was screened using the LncBase Predicted program, and the binding was verified by assaying the luciferase activity of a luciferase/lncRNA recombinant plasmid construct. The competitive endogenous RNA (ceRNA) relationship of the two RNAs was further validated by quantitative PCR after deregulation of each RNA using siRNA. The effect of miRNA and target genes on the MCF-7 cancer cell growth was determined by monitoring proliferation and apoptosis in the presence of Rh2 after deregulating the corresponding gene. The miRNA decreased the luciferase activity of the luciferase/CFAP20DC-AS1 fusion vector, confirming the binding. SiRNA-based deregulation of CFAP20DC-AS1 attenuated the expression of miR-3614-3p and vice versa. In contrast to CFAP20DC-AS1, miR-3614-3p was upregulated by Rh2, inhibiting proliferation but stimulating apoptosis of the MCF-7 cells. Target genes of miR-3614-3p, BBX and TNFAIP3, were downregulated by Rh2 and the miRNA but upregulated by the lncRNA. Rh2 inhibits CFAP20DC-AS1, which obscures the association of the lncRNA with miR-3614-3p, resulting in the suppression of oncogenic BBX and TNFAIP3. Taken together, the Rh2/CFAP20DC-AS1/miR-3614-3p/target gene axis contributes to the antiproliferation activity of Rh2 in cancer cells.
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Neoplasias da Mama , MicroRNAs , RNA Longo não Codificante , Apoptose/genética , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Ginsenosídeos , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , RNA Interferente Pequeno , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/genética , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/metabolismoRESUMO
We investigated whether (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone) could suppress the transcription factors expression and enzymes involved in glucose production by activating AMPK in hepatocytes. HepG2 cells were treated with a medium containing HM-chromanone (5-100 µM), compound C (10 µM) and insulin (100 nM). Glucose production and glycogen synthesis assay were determined using a glucose assay kit and glycogen assay kit, respectively. Activities of AMP-activated protein kinase (AMPK), acetyl CoA carboxylase (ACC), cAMP response element-binding protein (CREB), PPAR coactivator-1α (PGC1α), CREB-regulated transcription coactivator 2 (CRTC2), Glycogen synthase kinase (GSK3ß), Phosphoenolpyruvate carboxykinase (PEPCK), glycogen synthase (GS), Glucose 6-phosphatase (G6pase) and ß-actin were determined by Western blot analysis. HM-chromanone significantly inhibited hepatic glucose production and increased glycogen synthesis by activating glycogen synthase. HM-chromanone induced the phosphorylation of CRTC2 and GSK-3ß by phosphorylating AMPK in HepG2 cells, which was confirmed by compound C. Furthermore, it significantly decreased the phosphorylation of CREB in a time- and concentration-dependent manner, and the effect was reversed in the presence of compound C. Therefore, the complex formation of CRTC2 and CREB was inhibited. HM-chromanone inhibited the expression of PGC-1α, PEPCK, and G6Pase genes involved in production of hepatic glucose. The results showed that HM-chromanone activates AMPK in a time and concentration dependent manner, thus suppressing hepatic glucose production and increasing glycogen synthesis in HepG2 cells.
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Proteínas Quinases Ativadas por AMP , Glucose , Proteínas Quinases Ativadas por AMP/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Gluconeogênese , Glucose/metabolismo , Glicogênio/metabolismo , Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Células Hep G2 , Humanos , Hipoglicemiantes/farmacologia , Isoflavonas , Fígado/metabolismo , Fosfoenolpiruvato Carboxiquinase (ATP)/genética , FosforilaçãoRESUMO
The effects of (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HMC) on hyperglycemia and dyslipidemia were investigated in diabetic mice. Mice were separated into three groups: db/db, rosiglitazone and HMC. Blood glucose or glycosylated hemoglobin values in HMC-treated mice were significantly lower compared to db/db mice. Total cholesterol, LDL-cholesterol, and triglyceride values were lower, and HDL-C levels were higher, in the HMC group compared to the diabetic and rosiglitazone groups. HMC markedly increased IRS-1Tyr612, AktSer473 and PI3K levels and plasma membrane GLUT4 levels in skeletal muscle, suggesting improved insulin resistance. HMC also significantly stimulated AMPKThr172 and PPARα in the liver, and ameliorated dyslipidemia by inhibiting SREBP-1c and FAS. Consequently, HMC reduced hyperglycemia by improving the expression of insulin-resistance-related genes and improved dyslipidemia by regulating fatty acid synthase and oxidation-related genes in db/db mice. Therefore, HMC could ameliorate hyperglycemia and dyslipidemia in type 2 diabetic mice.
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Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Dislipidemias , Hiperglicemia , Resistência à Insulina , Animais , Glicemia/metabolismo , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/tratamento farmacológico , Dislipidemias/tratamento farmacológico , Hiperglicemia/tratamento farmacológico , Hiperglicemia/metabolismo , Hipoglicemiantes/farmacologia , Hipoglicemiantes/uso terapêutico , Insulina , Isoflavonas , Camundongos , Camundongos Endogâmicos C57BL , RosiglitazonaRESUMO
BACKGROUND: Tamoxifen (tam) is widely used to treat estrogen-positive breast cancer. However, cancer recurrence after chemotherapy remains a major obstacle to achieve good patient prognoses. In this study, we aimed to identify genes responsible for epigenetic regulation of tam resistance in breast cancer. METHODS: Methylation microarray data were analyzed to screen highly hypomethylated genes in tam resistant (tamR) breast cancer cells. Quantitative RT-PCR, Western blot analysis, and immunohistochemical staining were used to quantify expression levels of genes in cultured cells and cancer tissues. Effects of matrix metalloproteinase-1 (MMP1) expression on cancer cell growth and drug resistance were examined through colony formation assays and flow cytometry. Xenografted mice were generated to investigate the effects of MMP1 on drug resistance in vivo. RESULTS: MMP1 was found to be hypomethylated and overexpressed in tamR MCF-7 (MCF-7/tamR) cells and in tamR breast cancer tissues. Methylation was found to be inversely associated with MMP1 expression level in breast cancer tissues, and patients with lower MMP1 expression exhibited a better prognosis for survival. Downregulating MMP1 using shRNA induced tam sensitivity in MCF-7/tamR cells along with increased apoptosis. The xenografted MCF-7/tamR cells that stably expressed short hairpin RNA (shRNA) against MMP1 exhibited retarded tumor growth compared to that in cells expressing the control shRNA, which was further suppressed by tam. CONCLUSIONS: MMP1 can be upregulated through promoter hypomethylation in tamR breast cancer, functioning as a resistance driver gene. MMP1 can be a potential target to suppress tamR to achieve better prognoses of breast cancer patients.
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The conductive polymeric composites incorporating carbon nanotube (CNT) and carbonyl iron powder (CIP) have attracted much attention for various sensor applications. In this paper, a comprehensive study of the magneto-sensing property of a CNT-CIP embedded polymer composite is conducted to implement the composite as magneto-sensors. Thus, this study experimentally investigated the magneto-sensing performances of CNT-doped polymeric composites with the addition of CIP in terms of electrical conductivity, sensitivity, repeatability, and response time. First, the CNT-CIP clusters were manufactured and their interactions were analyzed with the zeta potential measurement and SEM observation. Then, the CNT-CIP clusters were embedded into the polymeric composites for the magneto-sensing evaluations. Experiments showed that the CNT contents in the range of percolation threshold (i.e., 0.5% and 0.75%) are optimal values for sensor applications. The addition of CNT 0.5% and 0.75% resulted in a high sensitivity of 7% and a faster response time within 400 ms. Experiment evaluation confirmed a high potential of implementing CNT-CIP composite as magneto-sensors.
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OBJECTIVES: In this study, we investigated whether scopoletin stimulated the secretion of insulin in pancreatic ß cells as well as the underlying mechanism involved in this process. METHODS: We incubated the INS-1 pancreatic ß cells with various concentrations of glucose (1.1, 5.6 or 16.7 mM) in the presence or absence of scopoletin. We then analysed the secretion of insulin in the cells treated with insulin secretion inhibitors or secretagogues. The intracellular influx of calcium induced by scopoletin was also analysed using the Fluo-2 AM dye. KEY FINDINGS: We found that scopoletin (1-20 µM) markedly induced the secretion of insulin in a glucose concentration-dependent manner compared with the control. At depolarizing concentrations of potassium chloride (KCl), scopoletin markedly enhanced the insulin secretion compared with the cells which were treated only with KCl. Moreover, the treatment with diazoxide-opening K+ATP channel and verapamil blocking Ca2+ channel significantly decreased the scopoletin-induced increase in insulin secretion. After the pre-treatment of cells with a Ca2+ fluorescent dye, treatment with 20 µM scopoletin resulted in a significant increase in the influx of intracellular Ca2+, exhibiting fluorescence changes in various spectra. CONCLUSIONS: Scopoletin stimulates the secretion of insulin via a K+ATP channel-dependent pathway in the INS-1 pancreatic ß cells.
Assuntos
Células Secretoras de Insulina , Ilhotas Pancreáticas , Trifosfato de Adenosina/metabolismo , Cálcio/metabolismo , Glucose/metabolismo , Insulina/metabolismo , Insulina/farmacologia , Células Secretoras de Insulina/metabolismo , Canais KATP , Escopoletina/metabolismo , Escopoletina/farmacologiaRESUMO
BACKGROUND: Ginsenoside Rh2, a major saponin derivative in ginseng extract, is recognized for its anticancer activities. Compared to coding genes, studies on long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) that are regulated by Rh2 in cancer cells, especially on competitive endogenous RNA (ceRNA) are sparse. METHODS: LncRNAs whose promoter DNA methylation level was significantly altered by Rh2 were screened from methylation array data. The effect of STXBP5-AS1, miR-4425, and RNF217 on the proliferation and apoptosis of MCF-7 breast cancer cells was monitored in the presence of Rh2 after deregulating the corresponding gene. The ceRNA relationship between STXBP5-AS1 and miR-4425 was examined by measuring the luciferase activity of a recombinant luciferase/STXBP5-AS1 plasmid construct in the presence of mimic miR-4425. RESULTS: Inhibition of STXBP5-AS1 decreased apoptosis but stimulated growth of the MCF-7 cells, suggesting tumor-suppressive activity of the lncRNA. MiR-4425 was identified to have a binding site on STXBP5-AS1 and proven to be downregulated by STXBP5-AS1 as well as by Rh2. In contrast to STXBP5-AS1, miR-4425 showed pro-proliferation activity by inducing a decrease in apoptosis but increased growth of the MCF-7 cells. MiR-4425 decreased luciferase activity from the luciferase/STXBP5-AS1 construct by 26%. Screening the target genes of miR-4425 and Rh2 revealed that Rh2, STXBP5-AS1, and miR-4425 consistently regulated tumor suppressor RNF217 at both the RNA and protein level. CONCLUSION: LncRNA STXBP5-AS1 is upregulated by Rh2 via promoter hypomethylation and acts as a ceRNA, sponging the oncogenic miR-4425. Therefore, Rh2 controls the STXBP5-AS1/miR-4425/RNF217 axis to suppress breast cancer cell growth.