Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP

Chronic kidney disease (CKD) progression is tightly associated with renal fibrosis, which is regulated by macrophage M2 polarization. The intestinal metabolite trimethylamine N-oxide (TMAO) has been reported to promote CKD, yet its underlying mechanism remains unclear. Here, we elucidated a mechanism wherein TMAO excreted through the kidneys alters the pyruvate metabolism of renal tubular epithelial cells, resulting in the production of lactic acid. Local lactic acid accumulation in the kidney promotes adjacent macrophage M2 polarization, a process speculated to be mediated by specific lactylation of macrophage genes. Through lactylation omics analysis, we identified histone H4 lysine 12 (H4K12) as the most significantly up-regulated lysine residue subjected to lactylation. Subsequent chromatin immunoprecipitation sequencing (ChIP-seq) assays revealed H4K12 lactylation on several glycometabolism gene promoters and genes. Furthermore, we found that this lactylation-mediated epigenetic regulation requires the assistance of the “porter”protein p300, as knockdown of p300 weakened the trend towards M2 polarization induced by lactic acid. Using an in vivo unilateral ureteral obstruction (UUO) mouse model, we verified the M2 polarization effect of TMAO and its detrimental role in CKD, as well as the protective effect of the TMAO inhibitor iodomethylcholine (IMC) on CKD. Clinical data validated the up-regulated TMAO’s effect on renal M2 polarization and fibrosis. Our findings suggest that CKD patients exhibit increased TMAO levels, which modulate the production of lactic acid by renal intrinsic cells. Epigenetic regulations mediated by lactic acid, particularly H4K12la on macrophage genes involved in glycometabolism, may contribute to M2 polarization. Targeting TMAO or its downstream pathways could have potential therapeutic benefits in CKD. Schematic diagram showing the whole TMAO modulation process. CKD dysfunction of microbiota leads to elevated TMA. TMA metabolized through liver into TMAO which excreted 90% through kidney. Renal tubular epithelial cells contact with TMAO and secrete lactic acid affecting adjacent macrophages more into M2 type through gene histone H4K12la under the help of p300 as a carrier. These genes include a large amount of glucose metabolism related genes which could at least partially explain this M2 polarization.

Spinal cord injury (SCI) is a severely disabling pathological condition and always results in sensory and motor functions loss. After SCI, inflammation-mediated secondary tissue damage and subsequent formation of neuroglial scar suppress neuron survival and regeneration, therefore resulting in severe neurofunctional dysfunction. Although nanoclay biomaterials demonstrated useful immunomodulation feature in non-neuronal tissue, it is unclear if they can inhibit inflammation in SCI lesions and promote neurofunctional recovery. In this study, we immobilized chondroitinase ABC enzyme (ChABC), which can degrade chondroitin sulfate proteoglycans (CSPGs) of neuroglial scar, inside nanoclay (Laponite®, Lap) hydrogel through electrostatic interaction. The presented ChABC-immobilized Lap hydrogel (ChABC@Lap) exhibited shear-thinning properties and excellent injectability, allowing to locally deliver it into injured area through a non-invasive manner. Cell biological experiments and SCI rat model research revealed that both pure Lap (without ChABC) and ChABC@Lap hydrogels significantly reduced pro-inflammatory macrophage marker (iNOS) intensity. Importantly, compared with Lap hydrogel, ChABC@Lap hydrogel significantly decreased GFAP and CS-56 marked scar deposition, increased Tuj-1 + neurons regeneration, and promoted NF200 + neurons survival, thereby improving SCI rat of electrophysiological characteristics and motor function. In summary, the designed enzyme-immobilized nanoclay hydrogel offers a promising strategy for SCI therapy through modulating simultaneously inflammation and glial scar deposition.

The immunosuppressive tumor microenvironment (ITME) promotes immune evasion and resistance to checkpoint blockade therapy. STING pathway activation offers a promising strategy to remodel the ITME, but existing agonists face limitations such as rapid degradation and poor bioavailability. Here, we developed natural compound rhein-based multifunctional bimetallic nanosheets (FGR NSs) composed of rhein (Rh), gadolinium (Gd 3+ ), and iron (Fe 3+ ) through a one-step symbiotic method to activate STING signaling and induce ferroptosis simultaneously. Upon decomposition in the tumor microenvironment, the natural product Rh triggers DNA double-strand breaks (dsDNA), promoting STING activation and increasing IFN-β secretion by 3.06-fold, while Fe 3+ drives ferroptosis through the Fenton reaction by catalyzing the conversion of endogenous hydrogen peroxide into highly toxic hydroxyl radicals and depleting glutathione (GSH), thereby promoting the generation of reactive oxygen species (ROS). Those mechanisms initiate both innate and adaptive immune responses and release abundant damage-associated molecular patterns (DAMPs, including CRT, ATP, HMGB1, and IFN-β) that promote dendritic cell maturation, CD8 + T-cell priming, and M2-to-M1 macrophage repolarization. In parallel, it suppresses the infiltration of immunosuppressive cells, including regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs). In vivo, FGR NSs elicit potent antitumor effects with 89.01 % tumor growth inhibition rates (TGI) in the subcutaneous breast cancer model. When combined with immune checkpoint inhibitor (αPD-1), a significant tumor inhibition in the lung metastatic model was achieved. Moreover, the Gd/Fe components enable T 1 /T 2 dual-mode MRI, positioning FGR as a promising theranostic platform for cancer immunotherapy.

Introduction Human beings and animals have been exposed to long-term artificial lighting environments to induce glucose metabolism disorder. Melatonin (MT) is involved in the regulation of glucolipid metabolism, and can prevent skeletal muscle wasting as well as sarcopenia-associated diseases. However, the effect of exogenous MT on skeletal muscle glucose metabolism and the involvement of the parasympathetic pathway have not been clarified. Objectives: To investigate the role of parasympathetic regulatory pathway in the mediating the effects of exogenous MT on skeletal muscle glucose metabolism following long-term light exposure. Methods: This study established rapid growth period broiler models, while characterized muscle histological analysis, glucose metabolism indexes and related genes expression through parasympathetic activation, exogenous MT administration and exogenous MT with parasympathetic inhibition experiments. Results: Long-term light exposure inhibited muscle glycogen synthesis, promoted muscle glycogen decomposition, increased anaerobic glycolysis, decreased aerobic respiration and induced the injury in breast muscle. Parasympathetic activation and exogenous MT caused a marked improvement in muscle glycogen accumulation, aerobic glycolysis and the injury in breast muscle. The exogenous MT beneficial functions were alleviated by parasympathetic inhibition. Furthermore, parasympathetic activation and exogenous MT administration protected cecal microbiota homeostasis, by improving stability of the gut microbiota community and increasing the relative abundance of Lactobacillus . Lactobacillus abundance was positively associated with muscle glycogen accumulation. Conclusion: Taken together, this study highlighted the role of the novel parasympathetic regulatory pathway in the effects of exogenous MT in maintaining glucose metabolism homeostasis and restoring the damage in skeletal muscle with long-term light exposure. The results indicate that gut microbiota are involved in the MT-parasympathetic regulatory network. This study filles the gap in autonomic nervous-endocrine regulation under long light exposure, and provides a new insight to alleviate long light exposure-induced glucose metabolism disorders to improve the growth and health of humans and animals.

Plain Language Summary Background: Sepsis-associated acute kidney injury (SA-AKI) drives high mortality in sepsis. The triggering receptor expressed on myeloid cells-1 (TREM1) plays critical roles in both infectious and non-infectious pathologies. However, the role of TREM1 in AKI still needs to be further clarified. Methods: Using both in vivo and in vitro experiments, we examined the role and underlying mechanism of TREM1 in AKI. Results: In this study, the level of sTREM in the urine of patients with SA-AKI was significantly higher than that of the healthy control group, although there was no significant difference in sTREM levels in the serum. In the SA-AKI mouse model, TREM1 deficiency markedly reduced serum creatinine levels in SA-AKI mice. Notably, TREM1 deficiency significantly promoted the expression levels of Il10 and Cd206 in the kidneys of SA-AKI mice. Cytometric Bead Array (CBA) analysis revealed that serum levels of the pro-inflammatory cytokines IL17A and IFN-γ were significantly diminished, whereas the anti-inflammatory factor IL10 was notably elevated. The ELISA results showed that the serum levels of CCL2 and CXCL1 in TREM1-deficient mice were significantly reduced. Mechanistically, experimental evidence indicated that TREM1 deficiency promoted M2 macrophage polarization by activating IRF4 via PI3K/AKT and STAT6 pathways. Conclusions: These findings confirmed that TREM1 is the primary regulatory factor of macrophage plasticity in SA-AKI, proposing a therapeutic strategy for the clinical intervention of SA-AKI-related kidney diseases. Plain Language Summary This study explored the role of TREM1 in sepsis-associated acute kidney injury (SA-AKI) using both human samples and mouse models. Researchers found that patients with SA-AKI had higher levels of sTREM in their urine compared to healthy individuals, but not in their serum. In mice, lacking TREM1 led to lower serum creatinine levels, indicating improved kidney function. TREM1 deficiency also increased anti-inflammatory markers and reduced pro-inflammatory cytokines, suggesting a shift towards a less inflammatory state. The study suggests that targeting TREM1 could be a promising strategy for treating SA-AKI by promoting beneficial macrophage activity in the kidneys. Text is machine generated and may contain inaccuracies. FAQ

Ferroptosis induction is particularly promising for cancer therapy when the apoptosis pathway is compromised. Current strategies in nanomedicine for inducing ferroptosis primarily focus on promoting the accumulation of reactive oxygen species (ROS). However, the presence of intracellular antioxidants, such as nuclear factor erythroid 2-related factor 2 (Nrf2), can limit the effectiveness of such therapy by activating detoxification systems and eliminating ROS. To overcome this challenge, we developed a synergistic ferroptosis-inducing agent by modifying manganese (Mn 2+ )–1,8-dihydroxy-3-hydroxymethyl-anthraquinone (aloe-emodin, AE) with polyvinyl pyrrolidone (PVP) to create nanoparticles (MAP NPs). In the tumor microenvironment, these NPs degraded and released AE and Mn(II), facilitating the generation of ROS and Mn(IV) through a Fenton-like reaction between hydrogen peroxide (H 2 O 2 ) and Mn(II). Mn(IV) subsequently interacts with glutathione (GSH) to induce a cyclic catalytic effect, and the depletion of GSH diminished the activation of glutathione-dependent peroxidase 4 (GPX4). Furthermore, AE inhibits the activity of Nrf2 and depleted GSH, thereby synergistically enhancing antitumor efficacy. Here it is demonstrated that MAP NPs effectively generate a robust ROS storm within tumor cells, suggesting that high-performance ferroptosis therapy is effective. Additionally, the inclusion of Mn(II) in the MAP NPs enables real-time monitoring of therapeutic efficacy via magnetic resonance T 1 -weighted contrast imaging.

Poly (adenosine 5′-diphosphate-ribose) polymerase inhibitors (PARPi) are increasingly important in the treatment of ovarian cancer. However, more than 40% of BRCA1/2-deficient patients do not respond to PARPi, and BRCA wild-type cases do not show obvious benefit. In this study, we demonstrated that progesterone acted synergistically with niraparib in ovarian cancer cells by enhancing niraparib-mediated DNA damage and death regardless of BRCA status. This synergy was validated in an ovarian cancer organoid model and in vivo experiments. Furthermore, we found that progesterone enhances the activity of niraparib in ovarian cancer through inducing ferroptosis by up-regulating palmitoleic acid and causing mitochondrial damage. In clinical cohort, it was observed that progesterone prolonged the survival of patients with ovarian cancer receiving PARPi as second-line maintenance therapy, and high progesterone receptor expression combined with low glutathione peroxidase 4 (GPX4) expression predicted better efficacy of PARPi in patients with ovarian cancer. These findings not only offer new therapeutic strategies for PARPi poor response ovarian cancer but also provide potential molecular markers for predicting the PARPi efficacy.

Background Functional dyspepsia (FD), characterized by complex pathophysiology and limited therapeutic options, is a prevalent gastrointestinal disorder. Wei-Dong Granules (WDGs) demonstrate significant clinical efficacy in FD treatment; however, their underlying mechanisms require elucidation. Purpose To investigate the therapeutic effects of WDGs on FD and delineate the associated molecular mechanisms. Methods An FD rat model was established using neonatal iodoacetamide-induced transient gastric injury followed by tail clamping and alternate-day fasting in adulthood. Rats received WDGs via gavage. Gastric motility (food intake, gastric emptying rate, residual food volume), behavioral symptoms, serum gastrointestinal hormones (Motilin, Gastrin, Ghrelin, CCK), and inflammatory factors were assessed. Gastric mucosal damage and repair were evaluated by H&E, Masson, TUNEL, and CD45 staining histologically. After that, UPLC-MS was carried out to identify chemical profiling of WDGs. Transcriptomics of gastric tissues and gut microbiota analysis of intestinal contents were performed, followed by multi-omics integration. ELISA, qPCR, immunohistochemistry, western blot, Masson staining, TUNEL staining, and CD45 staining were conducted to validate the proposed mechanism. Results WDG treatment significantly improved FD symptoms, evidenced by increased food intake and gastric emptying rate, decreased gastric residual volume, and alleviated behavioral abnormalities. WDGs mitigated gastric mucosal damage and promoted glandular and mucosal regeneration. It regulated gastrointestinal hormones (elevated Motilin, Gastrin, Ghrelin; decreased CCK) and reduced IL-6 levels. UPLC-MS identified 174 chemical components in WDG, with 132 confirmed by standards, predominantly flavonoids (50), organic oxides (26), and prenol lipids (23). Multi-omics analysis indicated that WDGs modulated gut microbiota dysbiosis (e.g., Bacteroides, Peptostreptococcus, Ruminococcus), promoted short-chain fatty acid (SCFA) production, activated the vagus nerve-hypothalamus Ghrelin pathway via the gut-brain axis, regulated gastrointestinal hormone secretion, enhanced antioxidant capacity, and facilitated gastric mucosal repair. Subsequent validation confirmed that WDGs alleviated FD through multi-targeted actions encompassing enhanced motility, anti-inflammation, antioxidant effects, and mucosal repair. Conclusion WDGs effectively treat FD by orchestrating gut microbiota-SCFA-gut-brain axis signaling, which enhances gastric motility, reduces inflammation and oxidative stress, and promotes mucosal repair. This integrated study elucidates the mechanism of WDGs and provides a scientific foundation for its clinical application in FD therapy.

Sugar substitutes that maintain the homeostasis of glucose, lipid, and protein metabolism are important for nutritional intervention in type 2 diabetes mellitus (T2DM). However, the specific metabolic effects remain unclear. The aim of this study was to systematically compare the effects of four common sugar substitutes on a high-fat diet (HFD) combined with a streptozotocin (STZ)-induced T2DM mouse model from the perspective of hepatic glucose, lipid, and protein metabolism. In this study, based on the establishment of a T2DM mouse model induced by an HFD combined with STZ and nontargeted metabolomics, the effects of four sugar substitutes on regulating and improving sugar, lipid, and protein metabolism were systematically evaluated. The results showed that mogroside V (MOG), stevioside (ST), and erythritol (ERT) enhanced protein synthesis via the mammalian target of the rapamycin/p-P70S6K pathway. MOG and ST also improved glucose and lipid metabolism by activating the phosphor-AMP-activated protein kinase (p-AMPK) pathway and upregulating peroxisome proliferator-activated receptor alpha/carnitine palmitoyltransferase 1. Sucralose primarily improves lipid metabolism by downregulating sterol regulatory element-binding protein 1, whereas ERT increases lipid droplet accumulation in the liver. These findings provide a foundation for the application of sugar substitutes in T2DM nutritional interventions.

Background Ambra1 has recently been identified as a key regulatory factor in the progression of mantle cell lymphoma (MCL). The objective of this study was to investigate the biological role and molecular mechanism of Ambra1 in MCL. Methods The m6A modification level of Ambra1 was detected by MeRIP-qPCR. Wild-type and mutant Ambra1 plasmids were constructed to verify the direct regulation of Ambra1 by METTL3-mediated m6A modification. The influence of METTL3/m6A/YTHDF2/Ambra1 on the viability, proliferation, migration, apoptosis, and cell cycle of MCL cells was evaluated by standard in vitro assays. RIP and RNA pull-down assays were performed to validate Ambra1 as a downstream target of YTHDF2. Xenograft tumor models were established using BALB/c nude mice to confirm the in vivo phenotype of METTL3 and Ambra1 silencing. Results Ambra1 was downregulated in MCL cells by METTL3-mediated m6A modification. Furthermore, knocking down METTL3 in the MCL cells inhibited their proliferation, migration, and invasion through the upregulation of Ambra1, while METTL3 overexpression had the opposite effect. The m6A reader protein YTHDF2 downregulated Ambra1 expression by binding to Ambra1-m6A. YTHDF2 knockdown inhibited the growth of MCL cells through Ambra1, while YTHDF2 overexpression had the opposite effect. Mechanistically, METTL3 downregulated Ambra1 in the MCL cells in an m6A-YTHDF2-dependent manner to inhibit apoptosis. Finally, METTL3 knockdown inhibited MCL progression in vivo by inducing Ambra1 expression. Conclusion METTL3 promotes MCL progression through YTHDF2-mediated degradation of Ambra1 mRNA, suggesting that the METTL3/YTHDF2/Ambra1 may serve as a potential therapeutic target for MCL.