Neutrophils and neutrophil-derived serine proteases (NSPs), including neutrophil elastase (NE) and proteinase 3 (PR3), are present in visceral fat of individuals and rodents with obesity, yet their roles in energy metabolism remain elusive. Here we show that neutrophil infiltration and NSP activation impair visceral fat browning in response to β-adrenergic receptor stimulation or cold exposure in male mice. Genetic deletion or local pharmacological inhibition of NE with sivelestat rescued visceral fat browning and enhanced thermogenesis. Mechanistically, NE/PR3 directly suppresses beige adipogenesis by arresting cell cycle via CDK4/cyclin D1 downregulation, impairs beige adipocyte differentiation by degrading insulin-like growth factor binding protein-3, and indirectly promotes M1 macrophage polarization. Administration of sivelestat suppressed diet-induced neutrophil infiltration, enhanced cold-induced visceral fat browning and decreased visceral fat content in mice. These findings reveal an unexpected inhibitory role of NSPs in visceral fat browning and suggest the potential of repurposing sivelestat as an anti-obese drug.

Obesity is closely associated with chronic low-grade inflammation in adipose tissue, which drives systemic metabolic dysfunction. Tetrandrine (TET), a bisbenzylisoquinoline alkaloid from Stephania tetrandra, exhibits potent anti-inflammatory and immunomodulatory effects, yet its impact on obesity-associated metabolic disorders remains unclear. Administration of TET significantly attenuated body weight gain, adipose tissue mass, glucose intolerance, and hyperinsulinemia in diet-induced and genetic obese mouse models. Histological analyses of adipose tissues revealed that TET treatment decreased adipocyte hypertrophy, macrophage infiltration, and fibrosis, which were corroborated by transcriptomic and protein-level assessments showing downregulation of pro-inflammatory cytokines and inhibition of NF-κB signaling. In vitro, TET suppressed NF-κB activation in macrophages without affecting adipocyte differentiation, indicating macrophages as one of potential cellular targets. Collectively, these findings support TET as a promising immunometabolic regulator capable of restoring adipose tissue homeostasis and improving systemic metabolic health, providing evidence for the therapeutic potential of natural bioactive compounds in obesity-related metabolic disorders.

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related mortality globally. Increasing evidence suggests that long non-coding RNAs play a critical role in cancer development, with the small nucleolar RNA host gene family being a key participant in multiple types of carcinogenesis, including HCC. Small nucleolar RNA host gene 1 (SNHG1) is a significant member of the SNHG family. SNHG1 expression consistently increases in various HCC-associated processes, such as cell proliferation, apoptosis, angiogenesis, migration, invasion, and treatment resistance. Higher SNHG1 expression levels predict worse prognosis by positively correlating with clinicopathological features, including larger tumour size, poor differentiation, and advanced stages in patients with HCC. Nevertheless, the precise role of SNHG1 in the initiation and progression of HCC remains unclear. Therefore, this review aims to summarise the current investigations on the pathogenesis of SNHG1 in HCC, highlighting its potential as a molecular marker for early prediction and prognostic assessment. As a multifunctional modulator, SNHG1 is extensively involved in molecular signalling pathways in HCC progression and is valuable for therapeutic targeting.

Current predictive biomarkers for clinical outcomes and treatment in hepatocellular carcinoma (HCC) are not reliable enough. Neddylation, a novel post-translational modification, plays a crucial role in the immunomodulation, metabolism, and pathogenesis of HCC. However, whether it can function as a powerful predictive biomarker for HCC remains unknown. In current research, we first identified NAE1 as the most significant neddylation-related gene affecting the prognosis of HCC patients mainly through weighted gene co-expression network (WGCNA) and machine learning. Subsequently, we determined NAE1 expression as an independent risk factor for HCC using univariate and multivariate Cox regression and constructed a nomogram integrating NAE1 expression with clinical characteristics to predict survival probabilities in HCC patients. Bulk and single-cell RNA sequencing analyses revealed that NAE1 expression was primarily positively connected with immune cell infiltration in HCC, as assessed by the six latest immune algorithms. In addition, drug sensitivity and molecular docking collectively revealed the influence of NAE1 expression on the IC50 values of the four agents and the binding interactions between NAE1 protein and these drugs. Furthermore, we found that NAE1 depletion suppressed proliferation, migration, and invasion of HCC cells in vitro experiments. In conclusion, NAE1 protein holds considerable potential as a valuable biomarker for predicting clinical outcomes, immune landscapes, and drug sensitivity in HCC, as well as a promising therapeutic target.

Pharmacological benefits of glucagon like peptide-1 receptor agonist (GLP-1RA) were mainly contributed to body weight reduction. This study aims to identify the novel mechanisms by which liraglutide improves adipose tissue homeostasis through cyclooxygenase (COX)-2 signaling. Liraglutide's impact on metabolic parameters in vivo was assessed in high-fat diet (HFD)-induced obese mice received liraglutide (1 mg/kg/day) treatment. In vitro experiments were conducted with differentiated adipocytes, and non-targeted metabolomics sequenced to identify key metabolites and COX-2 involvement during cold exposure. Liraglutide improved metabolic insulin resistance, accompanied with reduction of body weight and individual fat weight. Liraglutide suppressed adipocyte hypertrophy whereas induced lipolytic activity, including upregulation of browning-related markers PGC1α, UCP1, and ATGL in obese adipose tissues and differentiated 3T3-L1 adipocytes. Metabolomics analysis revealed elevated activation of COX-2 signaling, including increasing levels of COX-2 signaling and prostaglandin levels in subcutaneous adipose tissue from liraglutide-treated obese mice. COX-2 inhibition abolished liraglutide's effects on adipogenesis and lipolysis, and impaired adaptive thermogenesis in response to cold. In conclusions, liraglutide suppressed adipocyte hypertrophy dependent on upregulation of COX-2 activity. Targeting COX-2 pathway in adipose tissue was one of clinical concerns when obese patient treated with GLP-1RA.

Purpose This study aimed to explore the characteristics of intestinal microflora polymorphism in postmenopausal women, and to determine the pathophysiological changes of gene polymorphism of intestinal flora and bone metabolism in postmenopausal osteoporosis (PMOP) patients. Methods A total of 104 postmenopausal women with PMOP or normal bone density were included. Lifestyle, hip T-score, bone metabolism indexes (25(OH)D, PTH, beta-CTX, PINP), intestinal mucous membrane barrier function (diamine oxidase, D-lactic acid, LPS), gene polymorphisms, and characteristics of gut microbiota were examined. Results Women with PMOP had reduced physical activity, less dietary protein and calcium intake, lower levels of 25(OH)D, hip T-score, and BMD, but PMOP group had increased total energy and fat intake, and higher levels of PTH, beta-CTX, diamine oxidase, D-lactic acid, and LPS (p < .05 for all), as compared with normal subjects. Analyses of the alpha- and beta-diversity of fecal microbiota indicated remarkably differences in postmenopausal women with or without PMOP. In details, individuals with PMOP had increased abundances of some genera (e.g. Roseburia and Bacteroides), but decreased abundances of some genera (e.g. Streptococcus and Dorea). Furthermore, use of a random forest model based on differential abundant taxa and ROC analysis could efficiently identify women with PMOP in the present cohort (AUC = 0.93). Conclusion The incidence of PMOP was closely associated with fecal microbial compositions and intestinal functional changes. The present findings supported potential applications of gut microbiome analysis for early diagnosis of PMOP, and provided potential therapeutic targets.

Liver cirrhosis remains a major global health challenge, with liver transplantation currently representing the most effective treatment. Elucidating its molecular pathogenesis is therefore critical for the development of novel therapeutic strategies. Emerging evidence indicates that mitophagy plays a pivotal role in cirrhosis progression. In this study, we employed integrative bioinformatics to explore the association between mitophagy and liver cirrhosis, aiming to identify potential therapeutic targets. Differentially expressed genes (DEGs) were obtained from GSE77627 and GSE139602 data sets, followed by functional enrichment analysis. Mitophagy-related differentially expressed genes (mito-DEGs) were screened and assessed through receiver operating characteristic (ROC) analysis, immune cell infiltration profiling, and protein-protein interaction (PPI). Weighted gene coexpression network analysis (WGCNA) identified key modules, and intersecting genes with mito-DEGs highlighted RAB7B as a candidate hub gene, which was validated in external data sets. Experimental verification confirmed elevated RAB7B expression in activated hepatic stellate cells (HSCs) and a mouse model of liver cirrhosis. Functional assays demonstrated that RAB7B inhibition attenuated HSC activation, and molecular docking revealed strong binding affinity between RAB7B and predicted anticirrhosis compounds. We found that RAB7B knockdown restored TGF-β-induced mitophagy inhibition, enhanced mitochondria-lysosome colocalization, and showed dynamic regulation by mitophagy status, indicating its role as a negative and responsive regulator of mitochondrial clearance. Collectively, our findings identify RAB7B as a mitophagy-related hub gene driving liver cirrhosis progression and provide novel insights into its therapeutic potential.

Renal tubule cells act as a primary site of injury in diabetic kidney disease (DKD), with dysfunctional mitochondrial quality control (MQC) closely associated with progressive kidney dysfunction in this context. Our investigation delves into the observed inactivation of yes-associated protein 1 (YAP1) and consequential dysregulation of MQC within renal tubule cells among DKD subjects through bioinformatic analysis of transcriptomics data from the Gene Expression Omnibus (GEO) dataset. Receiver operating characteristic curve analysis unequivocally underscores the robust diagnostic accuracy of YAP1 and MQC-related genes for DKD. Furthermore, we observed YAP1 inactivation, accompanied by perturbed MQC, within cultured tubule cells exposed to high glucose (HG) and palmitic acid (PA). This pattern was also evident in the tubulointerstitial compartment of kidney sections from biopsy-approved DKD patients. Additionally, renal tubule cell-specific Yap1 deletion exacerbated kidney injury in diabetic mice. Mechanistically, Yap1 deletion disrupted MQC, leading to mitochondrial aberrations in mitobiogenesis and mitophagy within tubule cells, ultimately culminating in histologic tubular injury. Notably, Yap1 deletion-induced renal tubule injury promoted the secretion of C-X-C motif chemokine ligand 1 (CXCL1), potentially augmenting M1 macrophage infiltration within the renal microenvironment. These multifaceted events were significantly ameliorated by administrating the YAP1 activator XMU-MP-1 in DKD mice. Consistently, bioinformatic analysis of transcriptomics data from the GEO dataset revealed a noteworthy upregulation of tubule cells-derived chemokine CXCL1 associated with macrophage infiltration among DKD patients. Crucially, overexpression of YAP1 via adenovirus transfection sustained mitochondrial membrane potential, mtDNA copy number, oxygen consumption rate, and activity of mitochondrial respiratory chain complex, but attenuated mitochondrial ROS production, thereby maintaining MQC and subsequently suppressing CXCL1 generation within cultured tubule cells exposed to HG and PA. Collectively, our study establishes a pivotal role of tubule YAP1 inactivation-mediated MQC dysfunction in driving DKD progression, at least in part, facilitated by promoting M1 macrophage polarization through a paracrine-dependent mechanism.

Aim: Adipose tissue (AT) dysfunction that occurs in both obesity and lipodystrophy is associated with the development of cardiomyopathy. However, it is unclear how dysfunctional AT induces cardiomyopathy due to limited animal models available. We have identified vacuolar H+-ATPase subunit Vod1, encoded byAtp6v0d1, as a master regulator of adipogenesis, and adipose -specific deletion of Atp6v0d1 (Atp6v0d1(AKO)) in mice caused generalized lipodystrophy and spontaneous cardiomyopathy. Using this unique animal model, we explore the mechanism(s) underlying lipodystrophy-related cardiomyopathy. Methods and Results: Atp6v0d1(AKO) mice developed cardiac hypertrophy at 12 weeks, and progressed to heart failure at 28 weeks. The Atp6v0d1(AKO) mouse hearts exhibited excessive lipid accumulation and altered lipid and glucose metabolism, which are typical for obesity- and diabetes -related cardiomyopathy. The Atp6v0d1(AKO) mice developed cardiac insulin resistance evidenced by decreased IRS -1/2 expression in hearts. Meanwhile, the expression of forkhead box O1 (FoxO1), a transcription factor which plays critical roles in regulating cardiac lipid and glucose metabolism, was increased. RNA-seq data and molecular biological assays demonstrated reduced expression of myocardin, a transcription coactivator, in Atp6v0d1(AKO) mouse hearts. RNA interference (RNAi), luciferase reporter and ChIP-qPCR assays revealed the critical role of myocardin in regulating IRS -1 transcription through the CArG-like element in IRS -1 promoter. Reducing IRS -1 expression with RNAi increased FoxO1 expression, while increasing IRS -1 expression reversed myocardin downregulation-induced FoxO1 upregulation in cardiomyocytes. In vivo, restoring myocardin expression specifically in Atp6v0d1(AKO) cardiomyocytes increased IRS -1, but decreased FoxO1 expression. As a result, the abnormal expressions of metabolic genes in Atp6v0d1(AKO) hearts were reversed, and cardiac dysfunctions were ameliorated. Myocardin expression was also reduced in high fat diet -induced diabetic cardiomyopathy and palmitic acid -treated cardiomyocytes. Moreover, increasing systemic insulin resistance with rosiglitazone restored cardiac myocardin expression and improved cardiac functions in Atp6v0d1(AKO) mice. Conclusion: Atp6v0d1(AKO) mice are a novel animal model for studying lipodystrophy- or metabolic dysfunction -related cardiomyopathy. Moreover, myocardin serves as a key regulator of cardiac insulin sensitivity and metabolic homeostasis, highlighting myocardin as a potential therapeutic target for treating lipodystrophy- and diabetes -related cardiomyopathy.

Maintenance of intestinal barrier function contributes to gastrointestinal homeostasis and therefore cardiovascular diseases. A number of studies show that intestinal permeability is affected by excessive inflammatory responses. Krüppel-like factor (KLF) 4 is one of the critical transcriptional factors, which controls multiple immune responses. In this study we investigated the role of KLF4 in regulating intestinal inflammation and permeability during the atherosclerotic process. Atherosclerotic model was established in ApoE −/− mice by feeding a high fat high cholesterol (HFHC) diet. We showed that colon expression levels of KLF4 and tight junction proteins were significantly decreased whereas inflammatory responses increased in atherosclerotic mice. Overexpression of colon epithelial Klf4 decreased atherosclerotic plaque formation and vascular inflammation in atherosclerotic mice, accompanied by remarkable suppression of intestinal NF-κB activation. We found that overexpression of epithelial Klf4 in atherosclerotic mice significantly increased intestinal tight junction expression and ameliorated endotoxemia, whereas replenishment of LPS abolished these benefits. Overexpression of Klf4 reversed LPS-induced permeability and downregulation of ZO-1 and Occludin in Caco-2 cells in vitro. HFHC diet stimulated the expression of epithelial microRNA-34a, whereas silence of epithelial Klf4 abolished the benefits of microRNA-34a sponge, a specific miR-34a inhibitor, on intestinal permeability and atherosclerotic development. A clinical cohort of 24 atherosclerotic patients supported colon KLF4/NF-κB/tight junction protein axis mediated intestine/cardiovascular interaction in patients with atherosclerosis. Taken together, intestinal epithelial KLF4 protects against intestinal inflammation and barrier dysfunction, ameliorating atherosclerotic plaque formation.

BACKGROUND:Induction of beige fat for grafting is an emerging transplantation strategy. However, safety concerns associated with pharmaceutical interventions limit its wider application. Moreover, because beige fat is a special type of fat with strong metabolic functions, its effect on the metabolism of recipients after grafting has not been explored in the plastic surgery domain. OBJECTIVES:The aim of this study was to explore whether cold-induced inguinal white adipose tissue (iWAT) transplantation has a higher retention rate and beneficial effects on recipient metabolism. METHODS:C57/BL6 mice were subjected to cold stimulation for 48 hours to induce the browning of iWAT and harvested immediately. Subsequently, each mouse received a transplant of 0.2 mL cold-induced iWAT or normal iWAT. Fat grafts and recipients' iWAT, epididymal adipose tissue, and brown adipose tissue were harvested at 8 weeks after operation. Immunofluorescence staining, real-time polymerase chain reaction, and western blot were used for histological and molecular analysis. RESULTS:Cold-induced iWAT grafting had a higher mean [standard error of the mean] retention rate (67.33% [1.74%] vs 55.83% [2.94%], P < .01) and more satisfactory structural integrity than normal iWAT. Histological changes identified improved adipose tissue homeostasis after cold challenge, including abundant smaller adipocytes, higher levels of adipogenesis, angiogenesis, and proliferation, but lower levels of fibrosis. More importantly, cold-induced iWAT grafting suppressed the inflammation of epididymal adipose tissue caused by conventional fat grafting, and activated the glucose metabolism and thermogenic activity of recipients' adipose tissues. CONCLUSIONS:Cold-induced iWAT grafting is an effective nonpharmacological intervention strategy to improve the retention rate and homeostasis of grafts. Furthermore, it improves the adverse effects caused by traditional fat grafting, while also conferring metabolic benefits.

PDHA1 was associated with metabolic reprogramming in tumor progression. However, the clinical value of PDHA1, especially for prediction of drug sensitivity in hepatocellular carcinoma (HCC), has not been fully investigated. In this study, we found that PDHA1 expression was higher in HCC tissues compared to normal tissues and was correlated with poor prognosis in HCC patients. PDHA1 expression was mainly positively associated with immune cell infiltration using the TIMER, XCell, MCPCOUNTER, CIBERSORT, EPIC, and QUANTISEQ algorithms, which was validated by single-cell RNA-sequencing analysis. We also discovered that PDHA1 expression was correlated with six immune checkpoint-related genes. Univariate and multivariate Cox regression analyses revealed that PDHA1 expression was an independent prognostic indicator for HCC patients, and the nomogram incorporating PDHA1 expression exhibited excellent predictive capacity. Furthermore, PDHA1 expression was positively linked to the sensitivity of 5-fluorouracil, gemcitabine, paclitaxel, and sorafenib, and the molecular docking analysis demonstrated their excellent binding affinity.

BackgroundNuclear erythroid 2-related factor 2 (Nrf2), a transcription factor, is critically involved in the regulation of oxidative stress and inflammation. However, the role of endothelial Nrf2 in atherogenesis has yet to be defined. In addition, how endothelial Nrf2 is activated and whether Nrf2 can be targeted for the prevention and treatment of atherosclerosis is not explored.MethodsRNA-sequencing and single-cell RNA sequencing analysis of mouse atherosclerotic aortas were used to identify the differentially expressed genes. In vivo endothelial cell (EC)-specific activation of Nrf2 was achieved by injecting adeno-associated viruses into ApoE-/- mice, while EC-specific knockdown of Nrf2 was generated in Cdh5CreCas9floxed-stopApoE-/- mice.ResultsEndothelial inflammation appeared as early as on day 3 after feeding of a high cholesterol diet (HCD) in ApoE-/- mice, as reflected by mRNA levels, immunostaining and global mRNA profiling, while the immunosignal of the end-product of lipid peroxidation (LPO), 4-hydroxynonenal (4-HNE), started to increase on day 10. TNF-α, 4-HNE, and erastin (LPO inducer), activated Nrf2 signaling in human ECs by increasing the mRNA and protein expression of Nrf2 target genes. Knockdown of endothelial Nrf2 resulted in augmented endothelial inflammation and LPO, and accelerated atherosclerosis in Cdh5CreCas9floxed-stopApoE-/- mice. By contrast, both EC-specific and pharmacological activation of Nrf2 inhibited endothelial inflammation, LPO, and atherogenesis.ConclusionsUpon HCD feeding in ApoE-/- mice, endothelial inflammation is an earliest event, followed by the appearance of LPO. EC-specific activation of Nrf2 inhibits atherosclerosis while EC-specific knockdown of Nrf2 results in the opposite effect. Pharmacological activators of endothelial Nrf2 may represent a novel therapeutic strategy for the treatment of atherosclerosis.

BACKGROUND Competitive endogenous RNA (ceRNA) is an innovative way of gene expression modulation, which plays a crucial part in neoplasia. However, the intricacy and behavioral characteristics of the ceRNA network in hepatocellular carcinoma (HCC) remain dismal. AIM To establish a cyclin dependent kinase inhibitor 2A (CDKN2A)-related ceRNA network and recognize potential prognostic indicators for HCC. METHODS The mutation landscape of CDKN2A in HCC was first explored using the cBioPortal database. Differential expression analysis was implemented between CDKN2Ahigh and CDKN2Alow expression HCC samples. The targeted microRNAs were predicted by lncBasev3.0, and the targeted mRNAs were predicted by miRDB, and Targetscan database. The univariate and multivariate analysis were utilized to identify independent prognostic indicators. RESULTS CDKN2A was frequently mutated and deleted in HCC. The single-cell RNA-sequencing analysis revealed that CDKN2A participated in cell cycle pathways. The CDKN2A-related ceRNA network-growth arrest specific 5 (GAS5)/miR-25-3p/SRY-box transcription factor 11 (SOX11) was successfully established. GAS5 was recognized as an independent prognostic biomarker, whose overexpression was correlated with a poor prognosis in HCC patients. The association between GAS5 expression and methylation, immune infiltration was explored. Besides, traditional Chinese medicine effective components targeting GAS5 were obtained. CONCLUSION This CDKN2A-related ceRNA network provides innovative insights into the molecular mechanism of HCC formation and progression. Moreover, GAS5 might be a significant prognostic biomarker and therapeutic target in HCC.

Background Hepatocellular carcinoma (HCC), which is featured with high morbidity and mortality worldwide, is a primary malignant tumor of the liver. Recently, there is a wealth of supporting evidence revealing that NK cell-related immune traits are strongly associated with the development of HCC, but the causality between them has not been proven. Methods Two-sample Mendelian randomization (MR) study was performed to probe the causal correlation between NK cell-related immune traits and HCC. Genetic variations in NK cell-related immune traits were extracted from recent genome-wide association studies (GWAS) of individuals with European blood lineage. HCC data were derived from the UK Biobank Consortium's GWAS summary count data, including a total of 372,184 female and male subjects, with 168 cases and 372,016 controls, all of whom are of European ancestry. Sensitivity analysis was mainly used for heterogeneity and pleiotropy testing. Results Our research indicated the causality between NK cell-related immune traits and HCC. Importantly, CD8 + NKT cells had protective causal effects on HCC (OR = 0.9996;95%CI,0.9993–0.9999; P = 0.0489). CD16 − CD56 caused similar effects on NK cells (OR = 0.9997;95%CI,0.9996–0.9999; P = 0.0117) as CD8 + NKT cells. Intercepts from Egger showed no pleiotropy and confounding factors. Furthermore, insufficient evidence was found to support the existence of heterogeneity by Cochran's Q test. Conclusion MR analysis suggested that low CD8 + NKT cells and CD16 − CD56 expression on NK cells were linked with a higher risk of HCC.

OBJECTIVE:This study aimed to develop and validate a novel nomogram for diagnosing gastric cancer (GC). METHODS:In this prospective analysis, 146 patients of Wenzhou Central Hospital were recruited for a GC group and a benign lesion group and were divided into a training set and an internal validation set in a ratio of 7:3. Clinical and analytical characteristics were collected and analyzed by logistic regression analysis. The performance of the predictive model was evaluated using the receiver operating characteristic curve, calibration curve, and decision curve analysis. RESULTS:There were 5 variables, namely albumin, carcinoembryonic antigen, carbohydrate antigen 125, creatinine, and small proline-rich protein 2A, that were identified as the final parameters for the developed model. In the training and internal validation sets, the area under the curve of the model was 0.968 and 0.979, respectively, showing good diagnostic performance. CONCLUSION:This study developed and validated a new nomogram based on 5 parameters. This model shows good diagnostic performance in distinguishing GC from benign lesion groups and has certain significance in clinical application.