RESUMEN
Glucose lowering independently reduces liver fibrosis in human nonalcoholic fatty liver disease. This study investigated the impact of diabetes on steatohepatitis and established a novel mouse model for diabetic steatohepatitis. Male C57BL/6J mice were fed a 60% high-fat diet (HFD) and injected with carbon tetrachloride (CCl4) and streptozotocin (STZ) to induce diabetes. The HFD+CCl4+STZ group showed more severe liver steatosis, hepatocyte ballooning, and regenerative nodules compared with other groups. Diabetes up-regulated inflammatory cytokine-associated genes and increased the M1/M2 macrophage ratios in the liver. Single-cell RNA sequencing analysis of nonparenchymal cells in the liver showed that diabetes reduced Kupffer cells and increased bone marrow-derived recruited inflammatory macrophages, such as Ly6Chi-RM. Diabetes globally reduced liver sinusoidal endothelial cells (LSECs). Furthermore, genes related to the receptor for advanced glycation end products (RAGE)/Toll-like receptor 4 (TLR4) were up-regulated in Ly6Chi-RM and LSECs in mice with diabetes, suggesting a possible role of RAGE/TLR4 signaling in the interaction between inflammatory macrophages and LSECs. This study established a novel diabetic steatohepatitis model using a combination of HFD, CCl4, and STZ. Diabetes exacerbated steatosis, hepatocyte ballooning, fibrosis, regenerative nodule formation, and the macrophage M1/M2 ratios triggered by HFD and CCl4. Single-cell RNA sequencing analysis indicated that diabetes activated inflammatory macrophages and impairs LSECs through the RAGE/TLR4 signaling pathway. These findings open avenues for discovering novel therapeutic targets for diabetic steatohepatitis.
Asunto(s)
Diabetes Mellitus , Enfermedad del Hígado Graso no Alcohólico , Ratones , Masculino , Humanos , Animales , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Células Endoteliales/metabolismo , Transcriptoma , Ratones Endogámicos C57BL , Hígado/metabolismo , Enfermedad del Hígado Graso no Alcohólico/patología , Cirrosis Hepática/patología , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patología , Dieta Alta en Grasa/efectos adversosRESUMEN
Muscle atrophy is the cause and consequence of obesity. Proteasome dysfunction mediates obesity-induced endoplasmic reticulum (ER) stress and insulin resistance in the liver and adipose tissues. However, obesity-associated regulation of proteasome function and its role in the skeletal muscles remains underinvestigated. Here, we established skeletal muscle-specific 20S proteasome assembly chaperone-1 (PAC1) knockout (mPAC1KO) mice. A high-fat diet (HFD) activated proteasome function by â¼8-fold in the skeletal muscles, which was reduced by 50% in mPAC1KO mice. mPAC1KO induced unfolded protein responses in the skeletal muscles, which were reduced by HFD. Although the skeletal muscle mass and functions were not different between the genotypes, genes involved in the ubiquitin proteasome complex, immune response, endoplasmic stress, and myogenesis were coordinately upregulated in the skeletal muscles of mPAC1KO mice. Therefore, we introduced an immobilization-induced muscle atrophy model in obesity by combining HFD and immobilization. mPAC1KO downregulated atrogin-1 and MuRF1, together with their upstream Foxo1 and Klf15, and protected against disused skeletal muscle mass reduction. In conclusion, obesity elevates proteasome functions in the skeletal muscles. PAC1 deficiency protects mice from immobilization-induced muscle atrophy in obesity. These findings suggest obesity-induced proteasome activation as a possible therapeutic target for immobilization-induced muscle atrophy.
Asunto(s)
Atrofia Muscular , Complejo de la Endopetidasa Proteasomal , Ratones , Masculino , Animales , Complejo de la Endopetidasa Proteasomal/metabolismo , Ratones Obesos , Atrofia Muscular/metabolismo , Músculo Esquelético/metabolismo , Obesidad/metabolismoRESUMEN
Selenoprotein P is upregulated in type 2 diabetes, causing insulin and exercise resistance. We have previously reported that eicosapentaenoic acid (EPA) negatively regulates Selenop expression by suppressing Srebf1 in H4IIEC3 hepatocytes. However, EPA downregulated Srebf1 long before downregulating Selenop. Here, we report additional novel mechanisms for the Selenop gene regulation by EPA. EPA upregulated Foxo1 mRNA expression, which was canceled with the ERK1/2 inhibitor, but not with the PKA inhibitor. Foxo1 knockdown by siRNA initiated early suppression of Selenop, but not Srebf1, by EPA. However, EPA did not affect the nuclear translocation of the FoxO1 protein. Neither ERK1/2 nor PKA inhibitor affected FoxO1 nuclear translocation. In summary, FoxO1 knockdown accelerates the EPA-mediated Selenop downregulation independent of SREBP-1c in hepatocytes. EPA upregulates Foxo1 mRNA via the ERK1/2 pathway without altering its protein and nuclear translocation. These findings suggest redundant and conflicting transcriptional networks in the lipid-induced redox regulation.
Asunto(s)
Diabetes Mellitus Tipo 2 , Ácido Eicosapentaenoico , Regulación hacia Abajo , Proteína Forkhead Box O1 , Hepatocitos , Humanos , Insulina , ARN Mensajero , Selenoproteína P , Proteína 1 de Unión a los Elementos Reguladores de Esteroles , EsterolesRESUMEN
In this study, we carried out a heterogeneous cytoplasmic lipid content screening of Neochloris oleoabundans microalgae by dielectrophoresis (DEP), using castellated glassy carbon microelectrodes in a PDMS microchannel. For this purpose, microalgae were cultured in nitrogen-replete (N+) and nitrogen-deplete (N-) suspensions to promote low and high cytoplasmic lipid production in cells, respectively. Experiments were carried out over a wide frequency window (100 kHz-30 MHz) at a fixed amplitude of 7 VPP. The results showed a statistically significant difference between the dielectrophoretic behavior of N+ and N- cells at low frequencies (100-800 kHz), whereas a weak response was observed for mid- and high frequencies (1-30 MHz). Additionally, a finite element analysis using a 3D model was conducted to determine the dielectrophoretic trapping zones across the electrode gaps. These results suggest that low-cost glassy carbon is a reliable material for microalgae classification-between low and high cytoplasmic lipid content-through DEP, providing a fast and straightforward mechanism.