RESUMO
Polycystic ovary syndrome is a very common disease of gynecological endocrine, accompanied by irregular menstruation, hyperandrogenism, metabolic abnormalities, reproductive disorders and other clinical symptoms, which seriously endangers women's physical and mental health, but its etiology and pathogenesis are not completely clear. Recently, the contribution of exosomes to the diagnosis and treatment of various diseases in the biomedical field has attracted much attention, including PCOS. Exosomes are extracellular vesicles secreted by cells, containing various biologically active molecules such as cell-specific proteins, lipids, and nucleic acids. They are important signaling regulators in vivo and widely participate in various physiopathological processes. They are new targets for disease diagnosis and treatment. Considering the important role of non-coding RNAs during the development and treatment of PCOS, this article takes exosomal miRNAs as the breakthrough point for elucidating the physiological functions and therapeutic potential of exosomes during the development and treatment of PCOS through analyzing the effects of exosomal miRNAs on ovarian follicle development, hormone secretion, oxidative stress, inflammatory response and insulin resistance, thus providing new research directions and theoretical basis for PCOS pathogenesis, clinical diagnosis and prognosis improvement.
RESUMO
Chlorogenic acid (CGA) is a powerful antioxidant polyphenol molecule found in many diets and liquid beverages, playing a preventive and therapeutic role in various diseases caused by oxidative stress and inflammation. Recent research has found that CGA can not only improve clinical symptoms in PCOS patients but also improve follicular development, hormone status, and oxidative stress in PCOS rats, indicating the therapeutic effect of CGA on PCOS. Notably, our previous series of studies has demonstrated the expression changes and regulatory mechanisms of HIF-1alpha signaling in PCOS ovaries. Considering the regulatory effect of CGA on the HIF-1alpha pathway, the present article systematically elucidates the therapeutic role and molecular mechanisms of HIF-1alpha signaling during the treatment of PCOS by CGA, including follicular development, steroid synthesis, inflammatory response, oxidative stress, and insulin resistance, in order to further understand the mechanisms of CGA effects in different types of diseases and to provide a theoretical basis for further promoting CGA-rich diets and beverages simultaneously.
Assuntos
Síndrome do Ovário Policístico , Humanos , Feminino , Ratos , Animais , Síndrome do Ovário Policístico/tratamento farmacológico , Síndrome do Ovário Policístico/metabolismo , Ácido Clorogênico/farmacologia , Ácido Clorogênico/uso terapêutico , Transdução de Sinais , Estresse OxidativoRESUMO
Irinotecan is a topoisomerase I inhibitor which has been widely used to combat several solid tumors, whereas irinotecan therapy can induce liver injury. Liver injury generally leads to tissue hypoxia, and hypoxia-inducible factor-1α (HIF-1α), a pivotal transcription factor, mediates adaptive pathophysiological responses to lower oxygen condition. Previous studies have reported a relationship between HIF-1α and autophagy, and autophagy impairment is a common characteristic in a variety of diseases. Here, irinotecan (50 mg/kg) was employed on mice, and HepG2 and L-02 cells were cultured with irinotecan (10, 20 and 40 µM). In vivo study, we found that irinotecan treatment increased final liver index, serum aminotransferase level and hepatic lipid accumulation. Impaired autophagic flux and activation of HIF-1α/BNIP3 pathway were also demonstrated in the liver of irinotecan-treated mice. Moreover, irinotecan treatment significantly deteriorated hepatic oxidative stress, evidenced by increased MDA and ROS contents, as well as decreased GSH-Px, SOD and CAT contents. Interestingly, protein levels of NLRP3, cleaved-caspase 1 and IL-1ß were enhanced in the liver of mice injected with irinotecan. In vitro study, irinotecan-treated HepG2 and L-02 cells also showed impaired autophagic flux, while HIF-1α inhibition efficaciously removed the accumulated autophagosomes induced by irinotecan. Additionally, irinotecan treatment aggravated lipid accumulation in HepG2 and L-02 cells, and HIF-1α inhibition reversed the effect of irinotecan. Furthermore, HIF-1α inhibition weakened irinotecan-induced NLRP3 inflammasome activation in HepG2 cells. Taken together, our results suggest that irinotecan induces liver injury by orchestrating autophagy via HIF-1α/BNIP3 pathway, and HIF-1α inhibition could alleviate irinotecan-induced lipid accumulation in HepG2 and L-02 cells, which will provide a new clue and direction for the prevention of side effects of clinical chemotherapy drugs.
Assuntos
Autofagia , Doença Hepática Crônica Induzida por Substâncias e Drogas , Subunidade alfa do Fator 1 Induzível por Hipóxia , Metabolismo dos Lipídeos , Animais , Camundongos , Autofagia/genética , Doença Hepática Crônica Induzida por Substâncias e Drogas/genética , Doença Hepática Crônica Induzida por Substâncias e Drogas/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Irinotecano/toxicidade , Metabolismo dos Lipídeos/genética , Proteínas de Membrana/metabolismo , Proteínas Mitocondriais/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLRRESUMO
Testes produce sperms, and gamete generation relies on a proper niche environment. The disruption of hierarchical regulatory homeostasis in Leydig or Sertoli cells may evoke a sterile phenotype in humans. In this study, we recapitulated type 2 diabetes mellitus by using a high-fat diet- (HFD-) fed mouse model to identify the phenotype and potential mechanism of diabetes-induced testicular impairment. At the end of the study, blood glucose levels, testosterone structure, testicular antioxidant capacity, and testosterone level and the expression of hypoxia-inducible factor- (HIF-) 1α, apoptosis-related protein cleaved-caspase3, and autophagy-related proteins such as LC3I/II, p62, and Beclin1 were evaluated. We found that long-term HFD treatment causes the development of diabetes mellitus, implicating increased serum glucose level, cell apoptosis, and testicular atrophy (P < 0.05 vs. Ctrl). Mechanistically, the results showed enhanced expression of HIF-1α in both Sertoli and Leydig cells (P < 0.05 vs. Ctrl). Advanced glycation end products (AGEs) were demonstrated to be a potential factor leading to HIF-1α upregulation in both cell types. In Sertoli cells, high glucose treatment had minor effects on Sertoli cell autophagy. However, AGE treatment stagnated the autophagy flux and escalated cell apoptosis (P < 0.05 vs. Ctrl+Ctrl). In Leydig cells, high glucose treatment was adequate to encumber autophagy induction and enhance oxidative stress. Similarly, AGE treatment facilitated HIF-1α expression and hampered testosterone production (P < 0.05 vs. Ctrl+Ctrl). Overall, these findings highlight the dual effects of diabetes on autophagy regulation in Sertoli and Leydig cells while imposing oxidative stress in both cell types. Furthermore, the upregulation of HIF-1α, which could be triggered by AGE treatment, may negatively affect both cell types. Together, these findings will help us further understand the molecular mechanism of diabetes-induced autophagy dysregulation and testicular impairment, enriching the content of male reproductive biology in diabetic patients.
Assuntos
Diabetes Mellitus Tipo 2 , Testículo , Camundongos , Animais , Humanos , Masculino , Estresse Oxidativo , Autofagia , Testosterona , Glucose/farmacologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/farmacologiaRESUMO
The PEPC family proteins are ubiquitous in various plants and play an important role in the process of photosynthetic carbon assimilation and have many non-photosynthetic biological functions. However, PEPC genes have not been reported in apple. In this study, the members of apple MdPEPC family were identified based on the new apple genome data by bioinformatics analysis, and their expression patterns in different tissues and the apple axillary bud transcriptome treated by decapitation and TDZ (cytokinin) were analyzed in order to explore the role of MdPEPC genes in apple axillary bud outgrowth. The results showed that 6 MdPEPC family members were identified in apple, which distributed on 6 different chromosomes, and had similar physicochemical characteristics. Phylogenetic tree and sequence alignment analysis showed that the MdPEPC could be divided into two subgroups (Group â and Group â ¡), in which four members in MdPEPC family were clustered into Group â , belonging to plant-type PEPCs. However, MdPEPC4 and MdPEPC5 were clustered into Group â ¡ with AtPPC4, belonging to bacterial-type PEPCs. There were 7 pairs of fragments repeats among MdPEPC members, but no tandem repeats existed. The promoter cis-acting element analysis showed that MdPEPC genes were not only affected by light and stress, but also regulated by multiple hormones. The expression profiles showed that all MdPEPCs except MdPEPC4 and MdPEPC5 were expressed in different apple tissues. Transcriptome data analysis showed that the expression levels of MdPEPC1 and MdPEPC3 were up-regulated after decapitation and TDZ treatment, whereas MdPEPC2 was significantly down-regulated at 48 h after treatments. In conclusion, MdPEPC1, MdPEPC2 and MdPEPC3 were selected as the candidate genes involved in axillary bud outgrowth regulation for further study.
Assuntos
Malus , Regulação da Expressão Gênica de Plantas , Malus/genética , Malus/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
In plants, ammonium (NH4+) is the main nitrogen source and acts as a physiological and morphological response signaling molecule. Melatonin and dopamine are associated with plant responses to abiotic stress. However, previous studies have rarely focused on nutrient stress, and the roles of melatonin and dopamine in the uptake and metabolism of nitrogen in plants remain unclear. In this study, we investigated the regulatory effects of melatonin and dopamine on nitrogen utilization efficiency in apple seedlings under two NH4+ concentrations (2 and 0.1 mM) by measuring plant growth, root system architecture, 15NH4+ content, and related enzyme activity and gene expression. Under low nitrogen supply, apple seedling growth slowed and showed marked reductions in biomass accumulation, chlorophyll content, and nutrient uptake. However, both melatonin and dopamine significantly improved plant growth, chlorophyll content, and root development and enhanced antioxidant enzyme activity. Exogenous application of melatonin or dopamine also promoted the absorption and accumulation of 15NH4+ and enhanced nitrogen metabolism-related enzyme activity. At the molecular level, melatonin and dopamine significantly increased the expression levels of nitrogen metabolism genes and transporter genes. Overall, these results suggest that melatonin and dopamine can relieve nutrient stress caused by low concentrations of NH4+ through regulating the absorption and metabolism of nitrogen.
Assuntos
Compostos de Amônio , Malus , Melatonina , Dopamina , Melatonina/farmacologia , Nitrogênio , PlântulaRESUMO
Non-alcoholic fatty liver disease (NAFLD) is an independent risk factor for acute liver injury caused by overuse of acetaminophen (APAP). Caveolin-1 (CAV1), a regulator of hepatic energy metabolism and oxidative stress, was found to have a protective effect against NAFLD in our previous study. However, it remains unclear whether CAV1 has a protective effect against APAP-induced hepatotoxicity in NAFLD. The aim of this study was to determine whether CAV1 inhibits oxidative stress through the AMPK/Nrf2/HO-1 pathway to protect the liver from fat accumulation exacerbated by APAP in NAFLD. In this study, seven-week-old C57BL/6 male mice (18-20 g) were raised under similar conditions for in vivo experiment. In vitro, L02 cells were treated with A/O (alcohol and oleic acid mixture) for 48 h, and APAP was added at 24 h for further incubation. The results showed that the protein expression of the AMPK/Nrf2 pathway was enhanced after CAV1 upregulation. The effects of CAV1 on fat accumulation, ROS, and the AMPK/Nrf2 anti-oxidative pathway were reduced after the application of CAV1-siRNA. Finally, treatment with compound C (an AMPK inhibitor) prevented CAV1 plasmid-mediated alleviation of oxidative stress and fat accumulation and reduced the protein level of Nrf2 in the nucleus, demonstrating that the AMPK/Nrf2/HO-1 pathway was involved in the protective effect of CAV1. These results indicate that CAV1 exerted a protective effect against APAP-aggravated lipid deposition and hepatic injury in NAFLD by inhibiting oxidative stress. Therefore, the upregulation of CAV1 might have clinical benefits in reducing APAP-aggravated hepatotoxicity in NAFLD.
RESUMO
Alcoholic fatty liver (AFL) is a disease characterized by the abnormal structure and dysfunction of hepatocytes caused by long-term, excessive drinking. Acetaminophen (APAP) is a commonly used painkiller, but it can aggravate lipid deposition in the liver and cause liver injury when used in fatty liver disease. Here, we investigated the effect of caveolin-1 (CAV-1), an intracellular stent protein, on the pathogenesis of APAP aggravated lipid deposition in AFL mice. This study shows that lipid accumulation was more severe in APAP groups than in alcohol-treated mice. The CAV-1 stent-like domain (CSD, 82-101 amino acids of caveolin-1), used to upregulate CAV-1 expression, could reduce lipid accumulation and activate autophagy in AFL mice treated with APAP. The levels of CAV-1 and autophagy-related proteins (LC3-II/I and Beclin-1) had decreased, whereas SREBP-1c had increased in A/O (alcohol and oleic acid) and APAP-co-treated L02 cells. CAV-1 small interfering RNA and CAV1-overexpressing plasmid were separately transfected into A/O and APAP co-treated L02 cells. When CAV-1 was downregulated, the levels of Pink-1, Parkin, and autophagy-related proteins (LC3-II/I and Beclin-1) were decreased, whereas SREBP-1c was increased. The opposite trend was observed when CAV-1 was overexpressed. The results show that CAV-1 reduced lipid accumulation in L02 cells and activated Pink-1/Parkin-related mitophagy. This study highlights the positive role of CAV-1 in APAP-increased lipid accumulation under the AFL status and provides a new understanding of the function of CAV-1 in the liver through mitophagy associated with the Pink-1/Parkin pathway.
Assuntos
Acetaminofen , Mitofagia , Animais , Caveolina 1 , Fígado Gorduroso Alcoólico , Hepatócitos , Camundongos , Ubiquitina-Proteína LigasesRESUMO
Owing to the avascular structure of the ovarian follicle, proliferation of granulosa cells (GCs) and development of follicles occur under hypoxia, which is obviously different from the cell survival requirements of most mammalian cells. We hypothesized that autophagy may exert an inhibitory effect on GC apoptosis. To decipher the underlying mechanism, we constructed a rat follicular development model using pregnant mare serum gonadotropin and a cell culture experiment in hypoxic conditions (3% O2). The present results showed that the autophagy level was obviously increased and was accompanied by the concomitant elevation of hypoxia inducible factor (HIF)-1α and BNIP3 (Bcl-2/adenovirus E1B 19kDa-interacting protein 3) in GCs during follicular development. The levels of Bax (Bcl2-associated X) and Bcl-2 (B-cell lymphoma-2) were increased, while the activation of caspase-3 exhibited no obvious changes during follicular development. However, inhibition of HIF-1α attenuated the increase in Bcl-2 and promoted the increase in Bax and cleaved caspase-3. Furthermore, we observed the downregulation of BNIP3 and the decrease in autophagy after treatment with a specific HIF-1α activity inhibitor (echinomycin), indicating that HIF-1α/BNIP3 was involved in autophagy regulation in GCs in vivo. In an in vitro study, we also found that hypoxia did not obviously promote GC apoptosis, while it significantly enhanced the activation of HIF-1α/BNIP3 and the induction of autophagy. Expectedly, this effect could be reversed by 3-methyladenine (3-MA) treatment. Taken together, these findings demonstrated that hypoxia drives the activation of HIF-1α/BNIP3 signaling, which induces an increase in autophagy, protecting GC from apoptosis during follicular development.
RESUMO
Paeonol is a natural phenolic compound and isolated as an active ingredient from Moutan Cortex. Paeonol derivative-6 (DPF-6) is a derivative of paeonol improved in water solubility and bioavailability. Previous studies have reported that paeonol possesses a variety of pharmacological activities, such as antioxidant and anti-inflammatory properties. Moreover, we have previously verified that DPF-6 has anti-inflammatory effects. However, the role and fundamental mechanism of DPF-6 in acute liver injury (ALI) was still unclear. In this study, we indicated that DPF-6 inhibited inflammation and the expression of TNF-α, IL-6 and IL-1ß in liver tissues and LPS-mediated L-02 cells, concomitant with the upregulated expression of ZEB2. More importantly, it was demonstrated that overexpression of ZEB2 inhibited the expression level of TNF-α, IL-6 and IL-1ß in LPS-mediated L-02 cells. In contrast, knockdown of ZEB2 increased the expression level of TNF-α, IL-6 and IL-1ß in LPS-mediated L-02 cells. Further studies showed that ZEB2 inhibited the inflammation cytokine secretion via JNK signaling pathway in L-02 cells. Taken together, all the above results indicate that DPF-6 increased the expression of ZEB2, consequently inhibited inflammation cytokine secretion through JNK signaling pathway, which may be utilized as a potential anti-inflammation monomeric compound in the treatment of ALI.
Assuntos
Anti-Inflamatórios/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Fígado/efeitos dos fármacos , Homeobox 2 de Ligação a E-box com Dedos de Zinco/metabolismo , Acetofenonas , Animais , Linhagem Celular , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Galactosamina , Humanos , Mediadores da Inflamação/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Lipopolissacarídeos/toxicidade , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Fosforilação , Transdução de Sinais , Homeobox 2 de Ligação a E-box com Dedos de Zinco/genéticaRESUMO
The inflammasome is widely acknowledged for its crucial role in the pathogenesis of cancers and many neurodegenerative, metabolic, and auto-inflammatory diseases in recent years. Multiple types of inflammasomes exist. However, nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is the most often investigated inflammasome and has come to limelight in recent studies. NLRP3 inflammasome is a multi-protein complex. Its activation can cause the cleavage of inactive pro-caspase-1 into activated caspase-1, that ultimately promotes the transformation of pro-interleukin (IL)-1ß and pro-IL-18 into biologically-active IL-1ß and IL-18, respectively. These processes lead to the local inflammatory responses and induce pyroptosis, causing disparaging effects. Recently, numerous studies have shown that NLRP3 inflammasome plays an important role in the pathogenesis of liver diseases, including non-alcoholic fatty liver disease, liver fibrosis, cirrhosis, and hepatocellular carcinoma. Liver diseases have become a severe health burden worldwide, and there is adequate evidence indicating that the regulation of NLRP3 inflammasome acts as a guard against hazard to liver. In this review, we provide a straightforward overview of NLRP3 inflammasome as well as several frequent liver diseases. We then discuss the contribution and regulation of NLRP3 inflammasome during the pathogenesis of liver diseases, which may provide an important indication for the prevention and treatment of various liver diseases.
RESUMO
Non-alcoholic fatty liver disease (NAFLD) is the most burgeoning chronic liver disease worldwide whose pathogenesis is complex and controversial. Here, we investigated the impact of caveolin-1 (CAV1), a scaffolding protein of caveolae for lipid homeostasis and endocytosis, on the pathogenesis of NAFLD. CAV1 and caveolae play crucial roles in the regulation of autophagy and hepatic energy metabolism. However, it remains unclear whether CAV1 could affect hepatic lipid metabolism by regulating autophagy. In this study, results showed that the expressions of CAV1 and autophagy-related proteins (Beclin1 and LC3-II/â ) were decreased, while the level of p62 was increased in HFD (high-fat diet) fed mice liver and in A/O (alcohol and oleic acid mixture) treated L02 cells, compared to the corresponding controls. In vivo study, upregulation of CAV1 with CAV1 scaffolding domain peptides (CSD, amino acids 82-101 of caveolin-1) could alleviate lipid accumulation and promote autophagy in NAFLD mice. In vitro study, CAV1 overexpression plasmid and its small interfering RNA were cultured with A/O treated L02 cells respectively. The results also demonstrated that CAV1 reduced lipid accumulation and promoted autophagy in L02 cells. Treatment with chloroquine, an inhibitor of autophagic degradation, abrogated CAV1 plasmid-mediated alleviation of lipid accumulation. Mechanistically, the inhibition of Akt/mTOR pathway was involved in the protective role of CAV1 in autophagy induction and lipid metabolism in NAFLD. Together, these results provided novel perception into the function of CAV1 in liver through autophagy and emphasized its positive role in NAFLD.
Assuntos
Autofagia , Caveolina 1/metabolismo , Metabolismo dos Lipídeos , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Animais , Linhagem Celular , Regulação para Baixo , Humanos , Fígado/enzimologia , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Transdução de SinaisRESUMO
During the luteinization after ovulation in mammalian ovary, the containing cells undergo an energy consuming function re-determination process to differentiate into luteal cells under avascular environment. Previous evidences have delineated the contribution of autophagy to the cell differentiation and the catabolic homeostasis in various types of mammalian cells, whereas few interest had been focused on the involvement of autophagy in the luteinization of granulosa cells during the formation of early corpus luteum. Herein, the present study investigated that expression and contribution of autophagy during granulosa cell luteinization and early luteal development through in vivo and in vitro experiments. The results clearly demonstrated that HIF-1α/BNIP3-mediated autophagy plays a vital role in the luteinization of granulosa cells during the early luteal formation in vivo and in vitro. In the neonatal corpus luteum, HIF-1α up-regulated BNIP3 expressions, which contributed to the autophagic initiation by disrupting beclin1 from Bcl-2/beclin1 complex and protected cells from apoptosis by curbing the skew of mitochondria balance under avascular niche. Notably, Inhibition of HIF-1α activity by echinomycin enhanced the levels of cytoplasmic cytochrome c and cell apoptosis in the nascent corpus luteum. These findings revealed that HIF-1α/BNIP3-mediated autophagy enabled the process of granulosa cell luteinization and protected the granulosa-lutein cells from further apoptosis under hypoxia niche. To our knowledge, the present study firstly clarified that HIF-1α/BNIP3-mediated autophagy contributes to the luteinization of granulosa cells during the formation of pregnant corpus luteum, which will help us further understanding the luteal biology and provide us new clues for the treatment of luteal insufficiency.
RESUMO
BACKGROUND: Acute myeloid leukemia (AML) is an aggressive and mostly incurable hematological malignancy with frequent relapses after an initial response to standard chemotherapy. Therefore, novel therapies are urgently required to improve AML clinical outcomes. 4-Amino-2-trifluoromethyl-phenyl retinate (ATPR), a novel all-trans retinoic acid (ATRA) derivative designed and synthesized by our team, has been proven to show biological anti-tumor characteristics in our previous studies. However, its potential effect on leukemia remains unknown. The present research aims to investigate the underlying mechanism of treating leukemia with ATPR in vitro. METHODS: In this study, the AML cell lines NB4 and THP-1 were treated with ATPR. Cell proliferation was analyzed by the CCK-8 assay. Flow cytometry was used to measure the cell cycle distribution and cell differentiation. The expression levels of cell cycle and differentiation-related proteins were detected by western blotting and immunofluorescence staining. The NBT reduction assay was used to detect cell differentiation. RESULTS: ATPR inhibited cell proliferation, induced cell differentiation and arrested the cell cycle at the G0/G1 phase. Moreover, ATPR treatment induced a time-dependent release of reactive oxygen species (ROS). Additionally, the PTEN/PI3K/Akt pathway was downregulated 24 h after ATPR treatment, which might account for the anti-AML effects of ATPR that result from the ROS-mediated regulation of the PTEN/PI3K/AKT signaling pathway. CONCLUSIONS: Our observations could help to develop new drugs targeting the ROS/PTEN/PI3K/Akt pathway for the treatment of AML.
Assuntos
Antineoplásicos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Retinoides/farmacologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação para Baixo , Fluorimunoensaio , Humanos , Leucemia Mieloide Aguda , PTEN Fosfo-Hidrolase/efeitos dos fármacos , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de SinaisRESUMO
Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease which affects millions of people worldwide. Acetaminophen (APAP) overdose is the leading cause of acute liver failure. In this study, APAP (50, 100, 200â¯mg/kg) were employed on mice fed with a high-fat diet, and APAP (2, 4, 8â¯mM) were cultured with L02 cells in the presence of alcohol and oleic acid. APAP treatment significantly aggravated hepatic lipid accumulation, increased the serum levels of triglyceride (TG), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and increased hepatic lipid accumulation in H&E and Oil red O staining results. Transmission electron microscopy (TEM) found fewer number of autophagosomes in APAP (100â¯mg/kg) treated group. Immunohistochemistry analysis showed the intensity of hepatic mTOR was increased and AMPK was decreased in 200â¯mg/kg APAP treated group. Western blot analysis showed APAP treatment decreased the levels of LC3-â ¡, Beclin1 and AMPK, while increased the levels of mTOR and SREBP-1c, respectively. In vitro study showed APAP treatment obviously increased TG activities in cell supernatant, and Oil red O staining had the same results. Western blot analysis demonstrated APAP treatment decreased the levels of LC3-â ¡, Beclin1 and AMPK, increased the levels of mTOR and SREBP-1c, but rapamycin treatment significantly reversed these effects of APAP. In conclusion, therapeutic dosages of APAP aggravates fat accumulation in NAFLD, the potential mechanism might be involved in inhibiting autophagy associated with the AMPK/mTOR pathway, and patients with NAFLD should use a lower dose of APAP.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Acetaminofen/efeitos adversos , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/patologia , Autofagia/efeitos dos fármacos , Hepatopatia Gordurosa não Alcoólica/patologia , Serina-Treonina Quinases TOR/metabolismo , Animais , Autofagossomos/efeitos dos fármacos , Autofagossomos/patologia , Peso Corporal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND: Acute myeloid leukemia (AML) is an aggressive and mostly incurable hematological malignancy with frequent relapses after an initial response to standard chemotherapy. Therefore, novel therapies are urgently required to improve AML clinical outcomes. 4-Amino-2-trifluoromethyl-phenyl retinate (ATPR), a novel all-trans retinoic acid (ATRA) derivative designed and synthesized by our team, has been proven to show biological anti-tumor characteristics in our previous studies. However, its potential effect on leukemia remains unknown. The present research aims to investigate the underlying mechanism of treating leukemia with ATPR in vitro. METHODS: In this study, the AML cell lines NB4 and THP-1 were treated with ATPR. Cell proliferation was analyzed by the CCK-8 assay. Flow cytometry was used to measure the cell cycle distribution and cell differentiation. The expression levels of cell cycle and differentiation-related proteins were detected by western blotting and immunofluorescence staining. The NBT reduction assay was used to detect cell differentiation. RESULTS: ATPR inhibited cell proliferation, induced cell differentiation and arrested the cell cycle at the G0/G1 phase. Moreover, ATPR treatment induced a time-dependent release of reactive oxygen species (ROS). Additionally, the PTEN/PI3K/Akt pathway was downregulated 24 h after ATPR treatment, which might account for the anti-AML effects of ATPR that result from the ROS-mediated regulation of the PTEN/PI3K/AKT signaling pathway. CONCLUSIONS: Our observations could help to develop new drugs targeting the ROS/PTEN/PI3K/Akt pathway for the treatment of AML.