Expression Of Pro-inflammatory Factors Research Articles

Uterine Myometrial Distension Augments the Production of Angiogenic and Proinflammatory Factors

We recently found that myometrial distension stimulates maternal uterine vascular remodeling, and hypothesized that this may be a previously unrecognized mechanism for inducing arterial growth during pregnancy. The aim of this study was to further characterize a recently developed surgical model in which medical-grade silicone was infused into one uterine horn of a non-gravid rat to induce acute myometrial stretch, followed by an additional, gradual distension due to the secretion of an exudate into the uterine lumen. Our objectives were to better understand the effects of stretch on the myometrium and to look for the expression of proangiogenic and proinflammatory factors that may stimulate uterine vascular remodeling. Morphometric analysis showed hypertrophy of the uterine corpus that was primarily due to axial growth since the myometrial cross-sectional area was unchanged due to a thinning of the uterine wall secondary to stretch. This finding was supported by significantly increased myometrial smooth muscle cell mitosis. There was also an increase in the concentration of myometrial elastin but not collagen. The analysis showed modest increases in neutrophils, activated and unactivated macrophages, and the proinflammatory cytokines RANTES, MIP-3α, GRO-KC, and TNFα. The most dramatic change was the extremely high level of VEGF in the exudate, which was increased >900× above circulating levels.

Oral colon-targeted responsive chitosan/pectin-based nanoparticles propels the application of tofacitinib in colitis therapy

Tofacitinib (Tof), a commercially available pan-Janus kinases inhibitor, is approved for the treatment of moderate to severe ulcerative colitis. However, its clinical application is limited due to dose-dependent systemic side effects. The present study aims to develop an efficient oral colon-targeted drug delivery systems using prebiotic pectin (Pcn) and chitosan (Csn) polysaccharides as a shell, with Tof loaded into a Bovine Serum Albumin (BSA) core, and improving it with chondroitin sulfate (Chs), thus constructing Tof@BSA-Chs-CP nanoparticles (NPs). Our results suggest that the pH-sensitive characteristics of the Pcn/Csn shell contribute to its capacity for attenuating absorption and systemic diffusion in the gastrointestinal tract, and exhibiting targeted localization at inflamed colonic sites in mice. Additionally, the gut microbiota-secreted polysaccharide-degrading enzyme acts as the triggering agent for Pcn/Csn shell degradation. In mice colitis models, we demonstrated that oral administration of Tof@BSA-Chs-CP NPs effectively ameliorated colitis and expedited its resolution by modulating the expression of pro-inflammatory cytokines and immune regulatory factors. Collectively, our synthetic NPs demonstrate the promising potential of Tof for the therapy of UC.

Evodiamine rescues lipopolysaccharide-induced cognitive impairment via C/EBP-β-COX2 axis-regulated neuroinflammation.

Neuroinflammation is a key driver of neurological disorders. Evodiamine (EVO), an alkaloid from the traditional Chinese herb Evodia rutaecarpa, possesses potent biological activities, notably anticancer and anti-inflammatory effects. This study investigates EVO's potential to attenuate LPS-induced neuroinflammation, focusing on identifying its therapeutic targets and mechanisms of action. EVO treatment significantly improved mitochondrial function and reduced oxidative stress in LPS-stimulated BV2 cells, while also lowering levels of pro-inflammatory factors (IL-6, NO, IL-1β) in brain organoids. In mice, EVO treatment alleviated behavioral abnormalities, especially in cognition and memory, and lowered hippocampal inflammation marker levels. To elucidate the critical mechanisms by which EVO exerts its anti-inflammatory effects, we analyzed LPS-induced inflammatory injury in BV2 cells and used transcriptomics to investigate whether EVO modulates the C/EBP-β signaling pathway. Further validation using si-C/EBP-β confirmed EVO's regulatory effect on the C/EBP-β-COX2 axis, showing that knockdown significantly reduced pro-inflammatory factor expression, thereby providing neuroprotection. Moreover, molecular docking and dynamics simulations confirmed a stable interaction between EVO and C/EBP-β, supporting its role in attenuating LPS-induced neuroinflammation. In summary, these findings suggest that EVO regulates inflammation-related pathways by targeting the C/EBP-β-COX2 axis, offering neuroprotective benefits and mitigating neuroinflammatory responses.

Goose Deoxycholic Acid Ameliorates Liver Injury in Laying Hens with Fatty Liver Hemorrhage Syndrome by Inhibiting the Inflammatory Response

Fatty liver hemorrhagic syndrome (FLHS) in laying hens is a nutritional and metabolic disease involving liver enlargement, hepatic steatosis, and hepatic hemorrhage as the primary symptoms. The syndrome is prone to occur during the peak laying period of laying hens, which has resulted in significant economic losses in the laying hen breeding industry; however, the specific pathogenesis of FLHS remains unclear. Our group and previous studies have shown that bile acid levels are significantly decreased during the development of fatty liver and that targeted activation of bile acid–related signaling pathways is beneficial for preventing and treating fatty liver. In this study, we generated a FLHS laying hen model by feeding hens a high-energy, low-protein diet, with goose deoxycholic acid (CDCA) given as an intervention. HE staining, fluorescence quantitative PCR, and ELISA were used to evaluate the effects of CDCA on pathological changes and inflammatory responses in the liver. The results showed that hepatic hemorrhage in FLHS laying hens was reduced after CDCA treatment. Furthermore, fat vacuoles and transaminase levels decreased significantly. In addition, expression levels of M1-type macrophage markers and polarization products were significantly reduced, and the expression of pro-inflammatory regulatory factors related to the JAK-STAT signaling pathway, LPS-TLR4-Myd88–NF-kB signaling pathway, and NLRP3 inflammasomes decreased significantly as well. Expression levels of M2-type macrophage markers and polarization products increased significantly, as did the expression of anti-inflammatory regulators related to the JAK-STAT signaling pathway. These results suggest that CDCA ameliorates liver injury in laying hens with FLHS by inhibiting macrophage M1-type polarization and the resulting pro-inflammatory response, thereby promoting M2-type macrophage polarization and an anti-inflammatory response.

CAVIN2 attenuates ventilator-induced lung injury in rats by MAPK/ERK1/2 signaling pathway.

Mechanical ventilation is an important treatment in medical treatment, but it may cause or aggravate lung injury, which is called ventilator-induced lung injury (VILI). Studies have shown that CAVIN2 plays an important role in regulating inflammatory responses and cell death. However, its functional mechanism in VILI remains unclear. This study explores the potential role and mechanisms of CAVIN2 in the pathogenesis of VILI. We constructed rat and cell models of VILI. Real-time quantitative polymerase chain reactions (qRT-PCR), Western blot (WB), immunohistochemistry (IHC) and immunofluorescence (IF) were used to detect CAVIN2, ERK1/2, p-ERK1/2, autophagy-associated marker proteins LC3II/I, Beclin1 and P62, and the expression level of pro-inflammatory factors IL-1β and IL-6. Hematoxylin and eosin (H&E) staining were used to evaluate the degree of pathological injury of lung tissue, and the lung permeability was evaluated by measuring the wet-dry ratio of lung tissue and the total protein content in bronchoalveolar lavage fluid (BALF). Molecular docking, co-immunoprecipitation and immunofluorescence were used to verify the binding of CAVIN2 and ERK1/2, and the regulatory mechanisms of both were investigated by rescue experiments. CAVIN2 expression was downregulated in VILI rat lung tissues and ATII cells, whereas p-ERK1/2 expression was up-regulated. Overexpressing CAVIN2 alleviated pathological damage in VILI rat lung tissues and reduced the expression of pro-inflammatory factors and autophagy-related marker proteins in both lung tissues and ATII cells. Conversely, knockdown of CAVIN2 led to increased expression of pro-inflammatory factors and autophagy-related marker proteins in ATII cells. Further mechanism studies showed that CAVIN2 binds to ERK1/2 and inhibits ERK1/2 phosphorylation. Conversely when treated with the p-ERK1/2 agonist Ro67-7476, the protective, anti-inflammatory, and anti-autophagic effects of overexpressing CAVIN2 in VILI rats and ATII cells were reversed. CAVIN2 can bind to ERK1/2 and inhibit the activation of MAPK/ERK1/2 signaling pathway to reduce inflammatory response and autophagy in VILI, thereby reducing lung injury. Therefore, CAVIN2 may be a potential intervention target to provide a new strategy for the treatment of VILI.

GH inhibits ALV-J replication and restricts cell cycle by activating PI3K/Akt signaling pathway

Growth hormone (GH) plays a crucial role in growth, sexual maturity, and immunity in chickens. Avian leukosis virus subgroup J (ALV-J) is an exogenous tumorigenic retrovirus that primarily induces immunosuppression, growth retardation, decreased egg production, tumors formation, and even death in chickens. Previous studies have suggested that GH is involved in the regulation of innate immunity and inflammation. However, the specific role of GH in response to ALV-J remains unclear. In this study, we observed a significant upregulation of GH protein expression in the plasma of ALV infected chickens, and a marked increase in GH mRNA in ALV-J infected cells. We found that lower gp85 expression correlated with higher GH expression in immune tissues, suggesting that GH may inhibit gp85 expression. Additionally, GH overexpression enhanced the expression of interferons (IFN-α, IFN-β), interferon-stimulating genes (Mx1, ASCL1, CH25H), and pro-inflammatory factors (Mx1, ASCL1, CH25H) in DF-1 cells infected with ALV-J. GH also affected the cell cycle by regulating the expression of cell proliferation-related genes (p21, PCNA, Cyclin B2, Cyclin D1, Cyclin D2) and cell apoptosis-related genes (p53, Fas, Cyct, Caspase-1, Caspase-3, Caspase-8). More importantly, we found that GH restricted cell proliferation and apoptosis, and inhibited the replication of ALV-J by activating the PI3K/Akt signaling pathway in DF-1 cells. In conclusion, these results indicate GH plays a role in the antiviral response against the replication of ALV-J, providing evidence of an interaction between GH and the innate immunity in chickens.

Bioadhesive supramolecular polymer/hyaluronic acid hydrogel with zinc ion and dexamethasone slow release for diabetic wound healing

Diabetic patients often struggle with wound healing and are at higher risk of infections, necessitating the development of a stretchable, adhesive hydrogel dressing with antibacterial and angiogenesis-promoting properties. In this study, we synthesized a series of adhesive, antibacterial and anti-inflammatory hydrogels using free radical polymerization with materials including methacrylated hyaluronic acid (HAMA), N-[tris(hydroxymethyl)methyl]acrylamide (THMA), and 3-(bis(pyridin-2-ylmethyl)amino)propyl methacrylate (DPAMA). By leveraging the strong affinity of zinc(II)-dipicolylamine coordination complexes for the phosphorylated groups in dexamethasone sodium phosphate (DMSP), Zn2+ and DMSP were successfully incorporated into the hydrogel. The results demonstrated that the hydrogels possessed excellent adhesiveness and mechanical properties, enabling them to adhere closely to the skin while remaining easily removable without causing trauma. Antibacterial tests demonstrated significant inhibitory effects against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), attributed to the slow release of Zn2+, which effectively suppressed bacterial growth. Additionally, the slow release of DMSP provided strong anti-inflammatory effects. The DHTDZ2 hydrogel, containing 1.5 mg/mL Zn2+ and 4 mg/mL DMSP, significantly accelerated the healing of full-thickness skin wounds. In vitro angiogenesis, immunofluorescence, and immuno-histochemical results further confirmed that the DHTDZ2 hydrogel promoted angiogenesis and reduced the expression of pro-inflammatory factors. In summary, the hydrogel is an effective wound healing dressing that can reduce wound infections and inflammation.

Shikonin hastens diabetic wound healing by inhibiting M1 macrophage polarisation through the MAPK signaling pathway.

Diabetes is an endocrine disorder characterized by abnormally elevated blood glucose levels. Diabetic patients often exhibit impaired wound healing capabilities, particularly in the lower limbs, which is one of the numerous complications of diabetes. This is a significant factor leading to recurrent inflammation, disability, and even amputation. The primary objective of this study is to explore the mechanism by which shikonin accelerates diabetic wound healing by modulating macrophage phenotypes, particularly its role in the MAPK signaling pathway. To this end, we used a diabetic rat model and analyzed the effects of shikonin on the wound healing process and macrophage polarization in both in vivo and in vitro experiments. Additionally, we used immunofluorescence staining and Western blot techniques to detect the expression levels of macrophage polarization markers and proteins related to the MAPK signaling pathway. The results verify that shikonin significantly accelerated wound healing in diabetic rats and inhibited the polarization of M1 macrophages, reducing the expression of pro-inflammatory factors, while promoting the polarization of M2 macrophages, increasing the expression of anti-inflammatory factors. This process was accompanied by the regulation of the MAPK signaling pathway, indicating that shikonin accelerates diabetic wound healing by regulating the MAPK signaling pathway to inhibit the inflammatory phenotype of macrophages, showing significant clinical application prospects.

Chronic intermittent hypobaric hypoxia alleviates early-stage posttraumatic osteoarthritis via NF-κB/Nrf2 pathway in mice

BackgroundPosttraumatic osteoarthritis (PTOA) is directly associated with early acute articular cartilage injury. Inhibition of cartilage destruction immediately following joint damage can effectively slow or prevent PTOA progression. Therefore, we sought to determine intervention targets and therapeutic strategies in the acute stage of cartilage injury. The benefits of chronic intermittent hypobaric hypoxia (CIHH) extend to various body tissues, but its impact on acute cartilage injury remains unclear. We selected PTOA initiation as the therapeutic window and administered CIHH treatment immediately following cartilage injury initiation to investigate its protective effect on cartilage and molecular mechanism changing with time-varying.MethodsThe non-invasive PTOA mouse model was established by applying a single rapid specific impact force to the right knee’s tibial plateau, initiating load-induced PTOA development, closely resembling the pathological changes in human diseases. Following loading, we inhibited cartilage destruction by treating mice immediately in a hypobaric chamber with a hypobaric hypoxia mimic at 5000 m altitude. Cohorts of mice subjected to distinct experimental conditions were monitored for 3, 7, 14 or 28 days. Safranin O-Fast Green staining, Immunohistochemistry, immunofluorescence, ELISA, and western blotting were performed to evaluate the therapeutic effects of CIHH on cartilage in vivo. The nuclear translocation of NF-κB p65 and Nrf2 were detected by immunofluorescence.ResultsThe results showed that inhibiting cartilage destruction using CIHH immediately following acute articular cartilage injury initiation delayed the progression of PTOA, decreased the Mankin score and suppressed the expression of proinflammatory factors, including iNOS, NO, TNF-α, and IL-1β. Meanwhile, immediate CIHH treatment reduced levels of the catabolic enzymes ADAMTS5 and MMP13 in the cartilage matrix, reversed degradation of Collagen II and COMP, and inhibited oxidative stress by decreasing ROS levels. Moreover, CIHH suppressed NF-κB signaling by activating the Nrf2 in vivo studies.ConclusionOur study demonstrated that immediate CIHH treatment following cartilage injury initiation can attenuate load-induced cartilage damage by activating Nrf2/HO-1 and inhibiting the NF-κB p65 signalling pathways to counteract oxidative stress and inflammatory reactions, enhance the metabolic balance of the cartilage matrix and delay cartilage degeneration. This treatment may represent a potential therapeutic strategy for limiting PTOA progression.

Mitochondrial dysfunction in pancreatic acinar cells: mechanisms and therapeutic strategies in acute pancreatitis.

Acute pancreatitis (AP) is an inflammatory disease of the pancreas and a complex process involving multiple factors, with mitochondrial damage playing a crucial role. Mitochondrial dysfunction is now considered a key driver in the development of AP. This dysfunction often presents as increased oxidative stress, altered membrane potential and permeability, and mitochondrial DNA damage and mutations. Under stress conditions, mitochondrial dynamics and mitochondrial ROS production increase, leading to decreased mitochondrial membrane potential, imbalanced calcium homeostasis, and activation of the mitochondrial permeability transition pore. The release of mitochondrial DNA (mtDNA), recognized as damage-associated molecular patterns, can activate the cGAS-STING1 and NF-κB pathway and induce pro-inflammatory factor expression. Additionally, mtDNA can activate inflammasomes, leading to interleukin release and subsequent tissue damage and inflammation. This review summarizes the relationship between mitochondria and AP and explores mitochondrial protective strategies in the diagnosis and treatment of this disease. Future research on the treatment of acute pancreatitis can benefit from exploring promising avenues such as antioxidants, mitochondrial inhibitors, and new therapies that target mitochondrial dysfunction.

Licorice and dried ginger decoction inhibits inflammation and alleviates mitochondrial dysfunction in chronic obstructive pulmonary disease by targeting siglec-1.

Chronic obstructive pulmonary disease (COPD) is a chronic respiratory disease. Licorice and dried ginger decoction (LGD) is traditional Chinese medicine prescription with multiple effects. Glycyrrhetinic acid (GA) is the main bioactive components of LGD, which has been proven to have a relieving effect on various inflammatory diseases. Siglec-1 is a cell surface sialoadhesin and has been confirmed to be overexpressed in COPD and facilitate inflammatory reaction. This study is aimed to probe the interaction between LGD, GA, and siglec-1. Cigarette smoke (CS) combined with lipopolysaccharide (LPS) treatment was utilized to construct a COPD rat model. Cigarette smoke extract (CSE) was utilized to induce alveolar macrophage NR8383 to construct a COPD cell model. HE staining was applied for measuring histopathological changes of COPD rats. Enzyme-linked immunosorbent assay (ELISA), reverse transcription real-time polymerase chain reaction (RT-qPCR), and western blot were applied for testing the concentrations and expressions of proinflammatory factors. High performance liquid chromatography-tandem mass spectrometry (HPLC-MS) analysis was utilized to determine the combination between siglec-1 and GA. JC-1 assay was utilized to evaluate mitochondrial function. Reactive oxygen species (ROS) production was tested by dichloro-dihydro-fluorescein diacetate (DCFH-DA) staining. LGD treatment notably alleviated lung injury and inflammatory response in COPD rats. In CSE-induced cells, LGD treatment suppressed the contents of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8. Sialic-acid-binding Ig-like lectin 1 (Siglec-1) expression induced by CS was decreased after LGD treatment. Furthermore, we proved that GA could target siglec-1 to regulate the inflammatory response in COPD rats and cells. Additionally, GA could reduce ROS production and alleviate mitochondrial dysfunction to suppress COPD progression. LGD inhibits inflammation and alleviates mitochondrial dysfunction in COPD by targeting siglec-1.

Thermosensitive, injectable, antibacterial glabridin liposome/chitosan dual network hydrogel for diabetic wound healing.

Thermosensitive hydrogels show great potential in healing diabetic wounds, but they are still challenged by the long healing time, risk of infectivity, and accumulation of melanin. Herein, a dual network hydrogel is designed, which consists of chlorogenic acid (CA) modified chitosan (CS) (CA@CS), poly(N-isopropylacrylamide) (PNIPAm), and glabridin liposomes (GL). The gelation transition temperature of the hydrogel is 32-34°C, which thus endows it with superior injectability at ambient temperature. Moreover, GL/PNIPAm/CA@CS also exhibits excellent biocompatibility, and can promote the growth of epidermal cells, and the healing of diabetic wounds. GL/PNIPAm/CA@CS can also control the immune reactions by enhancing the release of CD206, and decreasing the formation of CD86 and ROS, which further promotes the production of CD31 and VEGF, and reduces the expression of pro-inflammatory factors, thus aiding in the healing of diabetic wounds. Furthermore, GL/PNIPAm/CA@CS can also suppress the growth bacterial, which can thus decrease the wound microbiota levels and facilitate the recovery of diabetic wounds. More importantly, it can reduce melanin production by 80% due to the action of glabridin. Consequently, GL/PNIPAm/CA@CS shows significant promise in enhancing the wound healing in future and decreasing the accumulation of melanin.

Flaxseed Linusorb Alleviates Collagen-Induced Rheumatoid Arthritis in Rats via Inhibiting the TLR4/NF-κb/MAPK Signal Pathway and Modulating Gut Microbiota.

Flaxseed linusorb (FL) has gradually garnered widespread attention in recent years because of its intriguing bioactivities like anti-inflammation, antimelanogenesis, and even anticancer effects. Based on its proven in vitro anti-inflammatory activity and mechanism, it is supposed that FL may also exhibit an in vivo effect in treating and preventing rheumatoid arthritis (RA). Collagen-induced arthritis (CIA) rat models were established to investigate the potential therapeutic effect of FL, which were intervened with FL via gavage (50 and 100 mg/kg B.W.) and intraperitoneal injection (10 and 20 mg/kg B.W.). After FL treatment, RA clinical symptoms were significantly alleviated, including reduced toe swelling volume and mitigated bone damage in CIA rats. Moreover, a decline in the expression of pro-inflammatory factors (i.e., TNF-α, IL-1β, and IL-6) and RA-related proteins (i.e., MMP-3, COX-2, and 5-LOX) was observed to effectively block the TLR4/NF-κB/MAPK signaling pathway. In addition, FL was discovered to modulate the diversity and composition of intestinal microbiota in CIA rats, where the level of g_Parvibacte, g_Allobaculum, g_Enterococcus, and unclassified_o_Lactobacillales could be significantly increased, whereas the level of Gram-negative bacteria g_Parabacteroides, g_Parasutterella, and g_Paludicola was notably reduced. In conclusion, FL shows promise in RA treatment by inhibiting inflammatory pathways and regulating the gut microbiota.

Polarization and Characterization of M1 and M2 Human Monocyte-Derived Macrophages on Implant Surfaces.

Foreign body reaction (FBR), an immune-mediated complex healing process, plays a crucial role in integrating implants into the body. Macrophages, as the first line of immune system interaction with implant surfaces, play a bidirectional role in modulating the inflammation-regeneration balance. For a deep understanding and the evaluation of the reactions between implant materials and immune responses, reliable in vitro methods and protocols are pivotal. Among different in vitro models, primary monocyte-derived macrophages (MDMs) present an excellent model for investigating macrophage-implant interactions. We have implemented an experimental protocol to evaluate the polarization of MDMs into M1 (classically activated) and M2 (alternatively activated) macrophages on implant surfaces. We isolated blood monocytes from healthy donors and differentiated them into macrophages using macrophage colony-stimulating factor (M-CSF). Differentiated macrophages were cultured on implant surfaces and polarized into M1 and M2 subtypes. M1 polarization was achieved in the presence of interferon (IFN)-γ and lipopolysaccharide (LPS), while M2 polarization was performed in a medium containing interleukin (IL)-4 and IL-13. We evaluated macrophage phenotypes by Enzyme-linkedImmunosorbent Assay (ELISA), confocal laser scanning microscopy (CLSM), and quantitative real-time PCR (qRT-PCR) based on panels of secreted cytokines, cell surface markers, and expressed genes. The extracted RNA was transformed into complementary DNA (cDNA), and qRT-PCR was used to quantify mRNA related to M1 and M2 macrophages. Accordingly, M1 macrophages have been characterized by higher expression of proinflammatory Tumor necrosis factor (TNF-α) cytokine and CCR7 surface marker compared to M2macrophages, which exhibited higher levels of CD209 and CCL13. Consequently, CCR7 and CD209 were identified as specific and reliable markers of M1 and M2 macrophage subtypes by immunostaining and visualizing by CLSM. Further confirmation was achieved by ELISA detecting elevated TNF-ɑ level in M1 and increased CCL13 in M2 cells. The proposed markers and experimental setup can be used effectively to assess the immunomodulatory potential of implants.

Profiling Pro-Inflammatory Proteases as Biomolecular Signatures of Material-Induced Subcutaneous Host Response in Immuno-Competent Mice.

Proteases are important modulators of inflammation, but they remain understudied in material-induced immune response, which is critical to clinical success of biomedical implants. Herein, molecular expression and proteolytic activity of three distinct proteases, namely neutrophil elastase, matrix metalloproteinases, cysteine cathepsins (cathepsin-K and cathepsin-B) are comprehensively profiled, in the subcutaneous host response of immuno-competent mice against different biomaterial implants. Quantitative non-invasive monitoring with activatable fluorescent probes reveals that different microparticulate materials induce distinct levels of protease activity with degradable poly(lactic-co-glycolic) acid inducing the strongest signal compared to nondegradable materials such as polystyrene and silica oxide. Furthermore, protein expression of selected proteases, attributable to both their inactive and active forms, notably deviates from their activities associated only with their active forms. Protease activity exhibits positive correlations with protein expression of pro-inflammatory cytokines tumor necrosis factor α and interleukin 6 but negative correlation with pro-fibrotic cytokine transforming growth factor β1. This study also demonstrates the predictive utility of protease activity as a non-invasive, pro-inflammatory parameter for evaluation of the anti-inflammatory effects of model bioactive compounds on material-induced host response. Overall, the findings provide new insights into protease presence in material-induced immune responses, facilitating future biomaterial assessment to evoke appropriate host responses for implant applications.

Homocysteine affects macrophage polarization by altering m6A methylation of scavenger receptors CD209 and CD163L1

ABSTRACT Atherosclerosis is a chronic inflammatory disease characterized by fatty plaque deposits on artery walls. Elevated plasma homocysteine (Hcy) levels are an independent risk factor for atherosclerosis. Research on the mechanism by which Hcy promotes atherosclerosis has gradually turned to epigenetic inheritance, but the correlation between Hcy and m6A (N6-methyladenosine) modification has not been reported. In this study, MeRIP-seq was performed on macrophages and Hcy-treated macrophages. GO and KEGG analyses were used to perform functional analysis of differentially methylated genes. qRT-PCR and western blot were taken to determine the expression of CD209, CD163L1, proinflammatory, and anti-inflammatory factors. Flow cytometry was used to detect the proportion of M2 macrophages. The results showed that after Hcy treatment, the overall m6A methylation of macrophages was down-regulated, and 856 differential methylation peaks were annotated to 781 genes. These included CD209 and CD163L1, whose m6A methylation was inhibited after treatment with Hcy. In addition, mRNA and protein expressions of CD209 and CD163L1 were also inhibited after Hcy treatment. Overexpression of CD209 or CD163L1 prevents the Hcy-induced decrease in the proportion of M2 macrophages. This article identified changes in the modification level of m6A in macrophages by Hcy and revealed the possible mechanism by which Hcy induces macrophage polarization.

Exploring the mechanism of Phyllanthus urinaria L. against ulcerative colitis based on network pharmacology and in vivo experiments

ObjectiveThis study aims to explore the therapeutic effect and molecular mechanism of the aqueous extract of Phyllanthus urinaria L. (PUL) on ulcerative colitis (UC). MethodPUL was administered to UC mice induced by 2.25% DSS. The changes of body weight, DAI score and colon length were recorded during the experiment. HE staining was conducted for pathology analysis of the mice in each group. The contents of inflammatory cytokines in colon tissues were detected by Elisa. Additionally, UPLC-TOF-MS/MS method was employed to identify the main components of PUL. Then, the TCMSP database was utilized to explore the potential active ingredients and drug targets of PUL. The matching of drug targets and disease targets yielded cross targets, which were used to construct a PPI network in String. Key targets underwent GO and KEGG enrichment analysis. Finally, Western blotting was performed to verify the key proteins expressions in the predicted pathway of network pharmacology and the expressions of tight junction proteins in the colon tissue. ResultPUL has been found to effectively alleviate the symptoms such as weight loss, bloody stools, and colon shortening in mice with UC. Administration of PUL led to an increase in tight junction protein in the colonic tissue of mice, as compared to the model group. Elisa results revealed that PUL reduced the expression of pro-inflammatory factors in mice with UC. HE staining results show that PUL can alleviate colon tissue damage caused by UC. A total of 773 components were detected in the water extract of PUL, among which 26 components, including quercetin, epigallocatechin gallate and kaempferol, may possess anti-UC activity. Network pharmacology analysis suggested that PUL may play an anti-ulcerative colitis role by inhibiting TNF pathway. Finally, Western blotting results confirmed that the PUL inhibited the TNF pathway and effectively treated ulcerative colitis. ConclusionThese preliminary research results indicate that PUL has the potential to exhibit anti-inflammatory activity by inhibiting the TNF pathway, thereby alleviating symptoms associated with UC. Substances such as quercetin, epigallocatechin gallate and kaempferol in PUL are believed to play an important role in this process.

Medium- and long-chain triacylglycerols as enteral nutrition enhancing the nutritional status of postoperative rats undergoing chemotherapy by increasing the bioavailability of functional fatty acids.

Medium- and long-chain triacylglycerols (MLCTs) have been widely used in the field of pharmaceutical nutrition. However, the application of MLCT as enteral nutrition remains underexplored. Our previous in vitro digestion experiment revealed that MLCTs can improve the bioaccessibility of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in comparison with physically mixed oils (MOs). In this study, we showed that MLCTs promote enhance triglyceride synthesis and secretion in Caco-2 cells, as evidenced by significant upregulation of RNA transcription levels of FATP4, L-FABP, and PPARα (p<0.05) compared with MOs. Animal experiment results show that MLCTs can improve the energy metabolism and liver function, decrease NF-κB p65 and inflammatory cytokines, such as TNF-α, IL-6, and IL-8 of rats undergoing chemotherapy postoperatively. This was related to the elevated bioavailability and metabolic efficiency of functional fatty acids in MLCTs. Besides, RT-qPCR, western blot, and immunohistochemical analyses confirme that MLCTs downregulate the expression of proinflammatory factors by suppressing the liver NF-κB signal transduction pathway, thereby improving the nutritional status of rats.

Female prediabetic rats are protected from vascular dysfunction: the role of nitroso and sulfide signaling

BackgroundThe activity of perivascular adipose tissue (PVAT), a specific deposit of adipose tissue surrounding blood vessels, could contribute to sex differences in vascular tone control, particularly in dyslipidemic disorders; however, the mutual associations remain unclear. This study aimed to evaluate the relationships among sex, PVAT and vascular function in Wistar and hereditary hypertriglyceridemic (HTG) rats. Vasoactive responses of the isolated thoracic aorta with preserved or removed PVAT were compared in adult male and female Wistar and HTG rats, and the roles of nitric oxide (NO), hydrogen sulfide (H2S), cyclooxygenase (COX) and inflammatory signaling in vascular function were monitored in females.ResultsHTG rats were hypertensive, but females less than males. Increased 2-h glycemia was observed in HTG rats regardless of sex; however, HTG females exhibited better glucose utilization than males did. Females, independent of strain, had better preserved endothelial function than males did. PVAT inhibited endothelium-dependent relaxation in all the rats except HTG females. In HTG males, pathologically increased aortic contractility was noted; however, in HTG females, the contractile responses were lower, thus approaching physiological levels despite the pro-contractile action of COX products. In HTG females, NO contributed to endothelial function to a lesser extent than it did in controls, but the presence of PVAT eliminated this difference, which corresponded with increased NO synthase activity. Although increased protein expression of several proinflammatory factors (TNFα, IL-6, iNOS, and NfκB) was confirmed in the aortic and PVAT tissue of HTG females, the protein expression of factors regulating the adhesion and infiltration of monocytes (ICAM-1 and MCP-1) was decreased in PVAT. Moreover, in HTG females, unlike in controls, H2S produced by PVAT did not inhibit endothelial relaxation, and regardless of PVAT, endogenous H2S had beneficial anticontractile effects, which were associated with increased protein expression of H2S-producing enzymes in both aortic and PVAT tissues.ConclusionsDespite increased inflammation and the pathological impact of cyclooxygenase signaling in female HTG rats, protective vasoactive mechanisms associated with milder hypertension and improved endothelial function and contractility linked to PVAT activity were triggered. Sulfide and nitroso signaling represent important compensatory vasoactive mechanisms against hypertriglyceridemia-associated metabolic disorders and may be promising therapeutic targets in prediabetic females.

Potential application of matrine microneedles for the treatment of atopic dermatitis in joint skin

Atopic dermatitis (AD) commonly manifests in skin areas subjected to flexion, particularly joints. This underscores the importance of dissolving microneedles (DMNs) used in AD treatment having strong skin adhesion to prevent drug delivery interruption due to patch detachment during patient activity. Moreover, matrine (MAT), a prominent active ingredient derived from the natural plant Sophora flavescens, is known for its significant efficacy in treating skin inflammation, itching, and allergic diseases. Herein, we presented MAT-loaded DMNs (MAT-DMNs) specifically tailored for joint skin application in AD treatment. MAT-DMNs can rapidly dissolve and hydrate after application, demonstrating adhesive properties on ex vivo pig skin, rat abdominal skin, and finger joint skin, thereby ensuring sustained and uninterrupted drug delivery to the skin. In vitro evaluations revealed that MAT-DMNs exhibited 24-hour first-order release profiles with cumulative penetration rates of 78.70 ± 4.24 % and 77.14 ± 6.65 % for the low-dose and high-dose groups, respectively. Furthermore, these DMNs showed excellent skin compatibility and therapeutic efficacy in AD mice models. Specifically, the skin lesion score in MAT-DMNs group was significantly lower (2.67 ± 0.40) compared to the model group (11.60 ± 1.62) and the blank DMNs group (10.80 ± 1.67). MAT-DMNs effectively alleviated AD symptoms such as itching and inflammation by reducing epidermal thickening, decreasing mast cell infiltration, and lowering the expression of Th2 cytokines (IL-4 and IL-10) and pro-inflammatory factors (IL-1β, IL-6, and TNF-α) compared to the blank DMNs group (p < 0.01). These findings suggest that MAT-DMNs hold promise for clinical AD treatment, particularly for skin diseases commonly affecting joint areas.