P38 MAPK Research Articles

Ethanolic extract of Parkia speciosa pods exhibits antioxidant and anti-inflammatory properties in lipopolysaccharide-induced murine macrophages by inhibiting the p38 MAPK pathway

BackgroundParkia speciosa (PS) is commonly used in Southeast Asian cuisine and traditional medicine to treat diabetes, hypertension, dermatitis, and kidney diseases. PS has emerged as a subject of interest because of its potential antioxidation and anti-inflammatory properties. However, despite its historically long and wide usage, a comprehensive investigation of these properties in PS pods (PSp) have not been conducted. Aims of this studyThis study aimed to identify the phytochemical compounds in the ethanolic extract of PSp collected from Southern Thailand and assess whether PSp exhibit antioxidant properties and mitigate inflammation in a lipopolysaccharide (LPS)-induced RAW264.7 model. Materials and methodsThe ethanolic extract of PSp was comprehensively analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry (GC/MS) to identify its phytochemical constituents. To assess the antioxidant activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid (ABTS) assays were performed, and cytotoxicity was evaluated using the MTT assay. The effect of PSp on reactive nitrogen and oxygen species (RNS and ROS) was determined using a nitric oxide (NO) assay, and its effect on pro-inflammatory cytokines was assessed using enzyme-linked immunosorbent assay (ELISA) and real-time quatitvative polymerase chain reaction (qPCR). Morphological changes following treatment were observed using a microscope. Western blot analysis was performed to quantify MAPK pathway expression. ResultsPSp contain polyphenols, phytosterols, triterpenes, oxaloacetic acid, and unsaturated fatty acids. PSp demonstrated high antioxidant potential in scavenging free radicals and exhibited no cytotoxic effects on macrophages. Moreover, PSp effectively reduced NO release and inhibited pro-inflammatory cytokines such as IL1-β, TNF-α, and IL-6. PSp treatment induced notable morphological changes in macrophages, characterized by an increase in cell size and the presence of intracellular vacuoles. In addition, Western blot analysis showed the selective suppressive effect of PSp on the p38-MAPK pathway. ConclusionPSp possess strong antioxidant and anti-inflammatory properties, making it a potential therapeutic agent for the treatment of inflammatory disorders.

ω-9 monounsaturated fatty acids in Sapindus mukorossi seed oil enhance calcium deposition expression of Wharton’s jelly mesenchymal stem cells

Promoting osteogenesis is crucial to improve successful bone regeneration in bone tissue engineering. Several studies on osteogenesis have reported positive effects of ω-9 monounsaturated fatty acids (MUFA) on bone regeneration. This study examined the potential of ω-9 monounsaturated fatty acid abundant seed oil mechanically extracted from Sapindus mukorossi (S. mukorossi) fruit to improve osteogeneses. After showing the presence of ω-9 MUFA in S. mukorossi seed oil (SM oil) through GC-MS spectrum analysis, the proliferation and differentiation of human umbilical cord Wharton’s jelly mesenchymal stem cells (WJMSCs) under SM treatment was evaluated. Our results indicate that WJMSC differentiation was induced by adding an osteogenesis-induced medium combined with SM oil. A high level of calcium deposition expression in WJMSCs induced by SM oil appears to be due to the effect of oleic acid and eicosenoic acid, both of which are ω-9 monounsaturated fatty acids. In addition, we found major contributors of SM oil-promoted WJMSC osteogenesis to be extracellular signal-regulated kinase (ERKs), c-Jun N-terminal kinase (JNKs), and the p38 MAPK pathway. The enhancement of WJMSC osteogenesis via ERK/MAPK pathways as demonstrated by qPCR analysis indicates the promise of SM oil for stem cell-based bone tissue engineering applications.

Surface Molecularly Engineered Mitochondria Conduct Immunophenotype Repolarization of Tumor-Associated Macrophages to Potentiate Cancer Immunotherapy.

Reprogramming tumor-associated macrophages (TAMs) to an inflammatory phenotype effectively increases the potential of immune checkpoint blockade (ICB) therapy. Artificial mitochondrial transplantation, an emerging and safe strategy, has made brilliant achievements in regulating the function of recipient cells in preclinic and clinic, but its performance in reprogramming the immunophenotype of TAMs has not been reported. Here, the metabolism of M2 TAMs is proposed resetting from oxidative phosphorylation (OXPHOS) to glycolysis for polarizing M1 TAMs through targeted transplantation of mannosylated mitochondria (mPEI/M1mt). Mitochondria isolated from M1 macrophages are coated with mannosylated polyethyleneimine (mPEI) through electrostatic interaction to form mPEI/M1mt, which can be targeted uptake by M2 macrophages expressed a high level of mannose receptors. Mechanistically, mPEI/M1mt accelerates phosphorylation of NF-κB p65, MAPK p38 and JNK by glycolysis-mediated elevation of intracellular ROS, thus prompting M1 macrophage polarization. In vivo, the transplantation of mPEI/M1mt excellently potentiates therapeutic effects of anti-PD-L1 by resetting an antitumor proinflammatory tumor microenvironmentand stimulating CD8 and CD4 T cells dependent immune response. Altogether, this work provides a novel platform for improving cancer immunotherapy, meanwhile, broadens the scope of mitochondrial transplantation technology in clinics in the future.

The antiviral effect and potential mechanism of Houttuynia cordata thunb. (HC) against coxsackievirus A4

Ethnopharmacological relevanceHand, foot, and mouth disease (HFMD) is mainly caused by various of enteroviruses such as enterovirus 71 (EVA71), coxsackievirus A16 (CVA16), CVA6, and CVA10 in infants and children under 5 years old. During the past 5 years, CVA4 has become the dominant pathogen resulting in HFMD in China. However, there are no effective vaccines and antiviral drugs available. Houttuynia cordata Thunb (HC). is a Chinese herbal medicine eaten as vegetables for treating viral infection diseases, but whether HC has anti-CVA4 effect remains unclear. Aim of the studyIn this study, we want to investigate the antiviral activity of HC against CVA4 in vitro and in vivo and elucidate the potential mechanism of HC against CVA4. Materials and methodsMTT assay were used to evaluate the cytotoxicity of HC. Virus titers assay, CPE assay, violet staining and immunofluorescence were used to investigate the antiviral effect of HC against CVA4. A 13-day-old suckling mice model was established to evaluate the therapeutic efficacy of HC against CVA4 infection. Western blot, qRT-PCR and time-of-drug addition assay were performed to elucidate the potential mechanism of HC against CVA4 infection. ResultsMTT assay indicated the cytotoxicity concentration of HC on Vero cells and RD cells were more than 1 mg/ml, suggesting that the low cytotoxicity of HC. In vitro antiviral assay revealed that HC could dose-dependently prevent the CPE, suppress the release of newborn virus, and inhibit the replication of CVA4 by decreasing viral RNA transcription and protein expression with IC50 of 88.96 μg/mL. A time-of-addition assay showed that HC mainly exerted anti-CVA4 effect by inhibiting virus replication at the post-entry stage. In vivo results further demonstrated that HC could effectively prevent the lethal infection of CVA4 by promoting survival, improving clinical symptoms, prolonging the survival time, inhibiting excessive inflammatory responses, and reducing pathological injury in vivo. Mechanistic studies revealed inhibition of p38 MAPK and JNK pathway over-activation may be the primary mechanism of HC against CVA4 infection. ConclusionIn summary, our results for the first time demonstrated that HC not only effectively inhibited CVA4 replication, but also partially protected the lethal infection of CVA4 in vivo. Furthermore, pharmacological mechanism studies revealed that the primary mechanism of HC against CVA4 infection may be associated with its effect of inhibiting over-activation of p38 MAPK and JNK signaling pathways caused by enteroviruses. Our finding indicated that HC might be a potential innovative medicine for treating HFMD.

Effect of interleukin-17A on inflammatory mediator production in interleukin-1β-stimulated human dental pulp fibroblasts.

In response to pro-inflammatory cytokines such as interleukin (IL)-1β, dental pulp fibroblasts produce various inflammatory mediators, including IL-6, IL-8, CC chemokine ligand 20 (CCL20), and CXC chemokine ligand 10 (CXCL10), leading to the progression of pulpitis. IL-17/IL-17A (IL-17A) is a pro-inflammatory cytokine secreted by T helper (Th) 17 cells following their recruitment to inflamed sites; however, the roles of IL-17A during pulpitis remain unclear. The purpose of this study was to investigate the effect of IL-17A on IL-6, IL-8, CCL20 and CXCL10 production by human dental pulp fibroblasts (HDPFs) in vitro. IL-17A at a concentration of 100ng/ml induced the production of 10 times more IL-8 and 4 times more CXCL10, but not IL-6 and CCL20, compared to controls. Co-stimulation of HDPFs with IL-17A and IL-1β synergistically enhanced the production of IL-6, CCL20, IL-8 and CXCL10. IL-1β increased expression of IL-17 receptor/IL-17RA (IL-17R) on HDPFs. Moreover, the cell signal pathways of p38 mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) were more potently activated by simultaneous stimulation with IL-17A and IL-1β. These findings suggest that IL-17A participates in the progression of dental pulp inflammation through the enhanced production of inflammatory mediators in HDPFs.

Ursodeoxycholic acid alleviates fat embolism syndrome-induced acute lung injury by inhibiting the p38 MAPK/NF-κB signalling pathway through FXR

Acute lung injury (ALI) caused by fat embolism syndrome (FES) is a disease with high mortality. This study aimed to explore the roles of ursodeoxycholic acid (UDCA) in FES-induced ALI and its underlying mechanisms. An ALI mouse model was established by allografting mouse perinephric fat. For in vitro experiments, human pulmonary microvascular endothelial cells (HPMEC) were treated with FFAs. The effects of UDCA on the expression of farnesoid X receptor (FXR) and the inflammatory response in endothelial cells were investigated. UDCA significantly inhibited the inflammatory response and the expression of proinflammatory markers during FES-induced ALI. UDCA markedly decreased TNF-α and IL-1β expression in vitro. UDCA administration markedly upregulated FXR expression and significantly reduced the phosphorylation of p38 MAPK and NF-κB p65. Knock down FXR expression decreased the effect of UDCA in vivo. Furthermore, knock down FXR expression and overexpressing FXR increased and decreased the inflammatory response, respectively, in vitro. Moreover, administration of a p38 MAPK activator reversed the anti-inflammatory effect of FXR overexpression. UDCA ameliorated inflammation during FES-induced ALI by suppressing p38 MAPK/NF-κB signalling and activating FXR. These findings provide new evidence for the potential of UDCA for FES-induced ALI treatment.

Trophic transfer induced gut inflammation, dysbiosis, and inflammatory pathways in zebrafish via Artemia franciscana: A differential analysis of nanoplastic toxicity

Rising glbal population and plastic consumption have caused a dramatic increase in plastic waste, leading to micro- and nanoplastic ingestion by aquatic organisms and subsequent bioaccumulation in their tissues. This transfer to higher trophic levels raises nanoplastic concentrations and bioavailability, potentially harming organisms' health and development. This poses a risk to human health via seafood. To address these issues, this study assesses the toxicological impacts of nanoplastics (NPs) on brine shrimp (Artemia franciscana) and their trophic transfer to zebrafish. The research unveiled concentration-dependent bioaccumulation of NPs in zebrafish and Artemia franciscana (A. franciscana). Polystyrene nanoplastics (PS-NPs) exhibited higher accumulation in A. franciscana whereas PP-NPs showed greater accumulation in zebrafish gut. Histopathological analysis under PS-NPs exposure revealed significant tissue alterations, indicative of inflammatory responses and impaired mucosal barrier integrity. Gene expression analyses confirmed these findings, showing activation of the P38-MAPK pathway by PS-NPs, which correlated with increased inflammatory cytokines. Additionally, PE-NPs activated the JNK-MAPK pathway, while PP-NPs exposure triggered the NOD-like receptor signaling pathway. Moreover, the composition of gut microbiota shifted to a dysbiotic state, characterized by an increase in pathogenic bacteria in the PS-NPs and PP-NPs groups, elevating the risk of developing Inflammatory Bowel Disease (IBD). PS-NPs were regarded as the most toxic due to their lower stability and higher aggregation tendencies, followed by PP-NPs and PE-NPs. Taken together, the overall study highlighted the complex interactions between NPs, gut microbiota, and host health, emphasizing the importance of thoroughly assessing the ecological and physiological impacts of nanoplastic pollution.

Exogenous Reactive Oxygen Species Augments SMAD4 Expression And TGF-β Paradox in Human Breast Cancer

Background: The Transforming growth factor-β (TGF-β) functions to induce apoptosis, cell cycle arrest, and differentiation is central to sustaining tissue homeostasis and maintaining genomic stability. TGF-β normally, an effective tumor-suppressor that restricts the uncontrolled division of cells augments the development and progression of human malignancies when cytostatic activities of TGF-β are resisted by genetic and epigenetic events caused by tumorigenesis. This dichotomic nature of TGF-β during oncogenesis termed as “TGF-β Paradox,” persists to be the most crucial and puzzling query regarding its physio-pathological function and the role of cellular antioxidant status is highly interrelated which warrants more studies on the role of endogenous reactive oxygen species (ROS) in deciding epithelial-mesenchymal transition (EMT) process. The objective of the study was to check whether enhanced ROS augments the TGF-β pathway facilitating EMT. Methods: In vitro toxicity assay was performed to assess the appropriate concentration of hydrogen peroxide (H2O2) imparting oxidative stress. Comet assay and 8-OHdG (8-hydroxy-2’-deoxyguanosine) enzyme-linked immunosorbent assay (ELISA) were performed to check the extent of DNA damage and adduct production respectively. Mitogen-activated protein kinase (MAPK) p38 ELISA and mRNA gene expression analysis of TGF-β and SMAD were done to verify the effect of H2O2 on these signaling. Results: The objective of the study was to check whether enhanced ROS augments the TGF-β pathway facilitating EMT. Along with morphological alterations, a dose-dependent decrease in cell viability was seen at 300µM of H2O2 compared to 75µM. DCFDA labeling discovered the dose-dependent gradation of intracellular ROS generation and this was correlated to increased cellular DNA damage and DNA adduct production which was increased linearly with increasing H2O2 as evident with comet test and 8-OHdG ELISA. Significantly reduced MAPK p38 activity revealed by indirect ELISA analysis suggests lessened suppression of cell growth. Conclusions: The study establishes that higher intracellular ROS will facilitate the TGF-β paradox leading to epithelial mesenchymal transition which can adversely affect therapeutic strategies targeting EMT

The Potential Role of Artemisinins Against Neurodegenerative Diseases.

Artemisinin (ART) and its derivatives, collectively referred to as artemisinins (ARTs), have been approved for the treatment of malaria for decades. ARTs are converted into dihydroartemisinin (DHA), the only active form, which is reductive in vivo. In this review, we provide a brief overview of the neuroprotective potential of ARTs and the underlying mechanisms on several of the most common neurodegenerative diseases, particularly considering their potential application in those associated with cognitive and motor impairments including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and amyotrophic lateral sclerosis (ALS). ARTs act as autophagy balancers to alleviate AD and PD. They inhibit neuroinflammatory responses by regulating phosphorylation of signal transduction proteins, such as AKT, PI3K, ERK, NF-[Formula: see text]B, p38 MAPK, I[Formula: see text]B[Formula: see text]. In addition, ARTs regulate GABAergic signaling in a dose-dependent manner. Although they competitively inhibit the binding of gephyrin to GABAergic receptors, low doses of ARTs enhance GABAergic signaling. ARTs can also inhibit ferroptosis, activate the Akt/Bcl-2, AMPK, or ERK/CREB pathways to reduce oxidative stress, and maintain mitochondrial homeostasis, protecting neurons from oxidative stress injury. More importantly, ARTs structurally combine with and suppress [Formula: see text]-Amyloid (A[Formula: see text]-induced neurotoxicity, reduce P-tau, and maintain O-GlcNAcylation/Phosphorylation balance, leading to relieved pathological changes in neurodegenerative diseases. Collectively, these natural properties endow ARTs with unique potential for application in neurodegenerative diseases.

Alleviation of Copper-Induced Hepatotoxicity by Bergenin: Diminution of Oxidative Stress, Inflammation, and Apoptosis via Targeting SIRT1/FOXO3a/NF-κB Axes and p38 MAPK Signaling.

Despite its biological importance, excess copper induces organ damage, especially to the liver. Disruption of critical signaling cascades that control redox status, inflammatory responses, and cellular apoptosis significantly contributes to the copper-induced hepatotoxicity. The present work explored the hepatoprotective ability of bergenin against the copper-induced hepatotoxicity using male Wistar rats as a mammalian model. The results revealed that bergenin suppressed the copper-evoked histopathological changes and hepatocellular necrosis as indicated by decreased activity of the liver enzymes ALT and AST in the sera of the copper-intoxicated rats. It decreased hepatic copper content and the copper-induced oxidative stress as revealed by reduced lipid peroxidation and improved activity of the antioxidant enzymes thioredoxin reductase, glutathione peroxidase, catalase, and superoxide dismutase. Bergenin downregulated the inflammatory cytokines TNF-α and IL-6, and the inflammatory cell infiltration to the liver tissues. Additionally, it inhibited the copper-induced apoptosis as indicated by significant reduction in caspase-3 activity. At the molecular level, bergenin activated the antioxidant transcription factor FOXO3a, inhibited the nuclear translocation of the inflammatory transcription factor NF-κB, and suppressed the inflammatory signaling molecules p38 MAPK and c-Fos. Interestingly, bergenin improved the expression of the anti-apoptotic protein Bcl2 and reduced the pro-apoptotic protein BAX. Bergenin markedly enhanced the expression of the histone deacetylase protein SIRT1 that regulates activity of NF-κB and FOXO3a. Collectively, these findings highlight the alleviating activity of bergenin against the copper-induced hepatotoxicity via controlling oxidative stress, inflammation, and apoptosis potentially through upregulation of SIRT1, activation of FOXO3a along with suppression of NF-κB and p38 MAPK signaling.

Salvia Miltiorrhiza Injection Inhibited the Proliferation of AML Cells by Inducing Apoptosis through the p38MAPK Pathway.

The purpose of this study was to explore the antitumor effect and mechanism of Salvia miltiorrhiza injection (SMI) on acute myeloid leukemia (AML) cells in vitro and in vivo. Bioinformatics was used to detect c-Myc mRNA expression in AML patients in the Oncomine database. qRT‒PCR and western blotting were used to detect the mRNA and protein expression of c-Myc and HOXA5 in clinical samples. Different concentrations (6.25, 12.5, 25, 50 and 100 μg/mL) of SMI were added to KG1a and HL60 cells for 24, 48 and 72 h to determine the IC50 value of SMI. A CCK-8 assay was used to detect the effects of different concentrations of SMI and different treatment times on the proliferation of KG1a and HL60 cells. The indicated concentrations of SMI and SB203580 were used to treat KG1a and HL60 cells. The cell cycle distribution was determined by flow cytometry. The percentage of apoptotic cells was detected by Hoechst 33258 staining and flow cytometry. qRT‒PCR was performed to detect the mRNA expression of p38, c-Myc and HOXA5 in KG1a and HL60 cells. Western blotting was used to detect the protein expression of p38, p-p38, c-Myc, HOXA5, cCaspase 3 and cPARP in KG1a and HL60 cells. AutoDock software was used to analyze the molecular docking of the three main active components of SMI with c-Myc. AutoDock analysis revealed that the binding effect of molecular leisure was evaluated by binding energy, and a binding energy <-5 kcal/mol was considered good. SMI decreased the mRNA and protein expression of c-Myc and HOXA5. SMI significantly inhibited the proliferative activity of KG1a and HL60 cells and induced their apoptosis. However, SMI had no significant effect on the cell cycle distribution of KG1a and HL60 cells. With increasing SMI concentrations, the p-p38/p38 ratio increased, while the protein expression of c-Myc and HOXA5 decreased, and the protein expression of cCaspase and cPARP increased. However, SB203580 intervention in addition to SMI reversed these changes. Tanshinone IIA, cryptanshinone and salvianolic acid B can bind to multiple sites of c-Myc. In summary, SMI could be used for the treatment of acute leukemia, and its mechanism may be related to activation of the p38MAPK signaling pathway.

Plant-Derived Alkaloids as a Potential Source of Treatment for Colorectal Cancer over the Past Five Years: A Comprehensive Review.

The gastrointestinal cancer known as colorectal cancer (CRC) is caused by a variety of genetic and epigenetic alterations in the intestinal epithelium of the colon and rectum. It is becoming more common every year. In view of this significant progress, it is urgent and imperative for researchers to work more in this direction in order to improve this health situation that is a major concern for society. Certain phenomena, such as the development of resistance by certain cells as well as the failure of certain therapies, play a part in the significantly changed situation. However, plants have always been used for their therapeutic virtues due to the large number of compounds they contain. Among them, alkaloids (more than 20,000 alkaloids have been isolated from plants, of which about 600 are known to be bioactive), which are one of the most diverse and extensively investigated classes of compounds among natural products, can be consider as a promising approach with regard to their numerous biological activities in general and, in particular their activities against colorectal cancer. This work aims to undertake deeper research on the examination of alkaloids that can be used as lead compounds in the treatment of colorectal cancer. The databases used during the literature searches were Web of Science, PubMed/Medline, and Scopus. This methodology allowed us to obtain 11 studies and 24 alkaloids (axidimins A-D, tabersonine, 19R-hydroxytabersonine, 11-hydroxytabersonine, 11-methoxytabersonine, vandrikidine, fusiformine A, 3-oxotabersonine, 3-oxo-11-methoxytabersonine, melodinine W2, venalstonidine, scandine, (-)-larutienine A, solasonin, berbamine dihydrochloride, nitidine chloride, GB7 acetate, berberine, boldine, Worenine, and chaetocochin J). Axidimin C and axidimin D showed significant cytotoxic effects on CRC (HCT116 cells) with IC50 values of 5.3 and 3.9 μM, respectively, and they were more active than 5-fluorouracil and etoposide (IC50 = 6.4 and 10.6 μM, respectively) taken as references. These two compounds induced G2/M phase arrest in HCT116 cells by downregulating cyclin B1 and cdc2 expression. Subsequently, promoting apoptosis via modulation of Bax and Bcl-2 levels, they enhanced p38 MAPK expression, leading to G2/M cell cycle arrest and apoptosis in HCT116 cells. Chaetocochin J possess significant activity against three different CRC cell lines [RKO (0.5 μM < IC50 = 0.56 μM < 1.0 μM), HCT116 (0.5 μM < IC50 = 0.61 μM < 1.0 μM) and SW480 (0.5 μM < IC50 = 0.65 μM < 1.0 μM)]. The 21 remaining compounds have a moderate anti-colorectal cancer activity. Thus, we believe that axidimin C, axidimin D and chaetocochin J could be promising compounds to fight colorectal cancer cell carcinoma. Nevertheless, future analysis should be performed on the study of the toxicologies of axidimin C and axidimin D.

Allocryptopine Attenuates Inflammatory Responses in Microglial Cells Via TLR4-Dependent NF-κB and p38 MAPK Pathways.

Studies in the existing literature propose that allocryptopine possesses both antioxidant and anti-inflammatory properties, showcasing its neuroprotective effects by potentially mitigating oxidative stress and inflammation. This study aims to investigate the antioxidant and anti-inflammatory effects of allocryptopine on various targets and potential mechanisms that have not been previously explored in the literature. Initially, we used MTT and LDH methods to evaluate the effects of allocryptopine on cell viability in BV-2 cells exposed to LPS-induced damage. Subsequently, we evaluated the impact of allocryptopine on pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), other inflammatory mediators (Cox-2 and iNOS), and p38 MAPK genes and proteins through qRT-PCR and Western blot analyses. Also, we evaluated the impact of allocryptopine on NF-κB proteins (TLR4, MyD88, IκBα, p-p50, and p-p65) through ELISA assay. Molecular docking analyses were performed to investigate the potential binding of allocryptopine to target proteins (TLR4, MyD88, IκBα, p50, p65, MKK3, MKK4, MKK6, p38, AP-1 (c-Jun and ATF2), IL-1β, IL-6, TNF-α, Cox-2, and iNOS) associated with the TLR4, NF-κB, and p38 MAPK pathways. Our results indicate that allocryptopine exerts a comprehensive influence on pro-inflammatory cytokines and other inflammatory mediators by inhibiting TLR4 signaling and modulating the NF-κB and p38 MAPK pathways. The outcomes of our study suggest that the antioxidant and anti-inflammatory efficacy of allocryptopine is intricately linked to the modulation of key molecular pathways associated with oxidative stress and inflammation. These findings highlight the potential of allocryptopine as a therapeutic agent for addressing neurodegenerative diseases by safeguarding neuronal health.

Enhanced activation of signaling pathway by recombinant human adiponectin from genome-edited chickens

Adiponectin (ADPN) exerts various cellular and metabolic functions by activating signaling pathways, including extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) pathways, the protein kinase B (Akt) pathway, and the p38 mitogen-activated protein kinase (MAPK) pathway. However, generating functional recombinant human adiponectin (hADPN) in bacterial or mammalian cells is challenging. Although ADPN agonist peptides have been developed, problems like stability, solubility, and affinity for receptors remain. Recently, a genome-edited chicken bioreactor system was established, ensuring efficient ADPN production with optimal post-transcriptional modifications. We assessed the ability of egg white (EW)-derived hADPN, commercial hADPN, various ADPN agonist peptides, and globular ADPN on activation of the ERK1/2, Akt, and p38 MAPK pathways. EW-derived hADPN, abundant in hexamers and high molecular weight multimers, significantly phosphorylated ERK1/2 in serum-starved HEK293 cells after 15 min of treatment. Comparative analysis revealed that EW-derived hADPN and commercial hADPN induced greater phosphorylation of ERK1/2, Akt, and p38 MAPK than ADPN agonist peptides and globular ADPN, with EW-derived hADPN showing the highest activation. In summary, the finding that EW-derived hADPN strongly activates the ERK1/2, Akt, p38 MAPK signaling pathways highlights that an ADPN production system based on genome-edited chickens is an advantageous alternative to existing methods.

Lif-deficiency promote systemic Iron metabolism disorders and increases the susceptibility of osteoblasts to ferroptosis

Leukemia inhibitory factor (LIF) is a multifunctional cytokine that plays a crucial role in various biological processes. However, LIF involvement in iron metabolism remains almost unexplored. This study aimed to explore the impact of LIF on systemic iron transportation and its potential role in ferroptosis in osteoblasts. We observed that the Lif-deficient (Lif−/−) mice is characterized by a reduction in bone mass and a decrease in bone mineral density compared with wild-type (WT) mice. Energy-dispersive X-ray spectroscopy revealed a marked increase in iron content on the surface of femurs from Lif−/− mice. Meanwhile, iron stores test lower iron levels in the spleens and higher levels in the femurs of Lif−/− mice. Besides, Lif−/− mice display increased levels of serum iron, total iron-binding capacity, unsaturated iron-binding capacity, and transferrin saturation and serum ferritin relative to WT mice. Hepcidin mRNA expression reduction in the liver of Lif−/− mice. It also holds true in the AML-12 hepatocyte cell line after Lif-knockdown. Immunohistochemistry and RT-PCR revealed elevated ferroportin (FPN) in duodenal cells of Lif−/− mice. Lif-deficiency decreases SLC7A11 levels in osteoblasts. In addition, overexpression of LIF downregulates CD71, DCYTB, and DMT1, thereby reducing iron uptake in iron-overloaded cells. Femur immunohistochemistry (IHC) revealed increased ACSL4 and decreased GPX4 and SLC7A11, indicating an increase in ferroptosis of osteoblasts in Lif−/− mice. Whole-transcriptome sequencing showed gene expression changes after Lif-knockdown, exhibiting a negative correlation with genes involved in long-chain fatty acid transport, mitochondrial organization, and the p38 MAPK signaling pathway. These results demonstrate that Lif-deficiency alter systemic iron metabolism and increases the susceptibility of osteoblasts to ferroptosis.

Structural characterization and immunoregulatory mechanism of a low-molecular-weight polysaccharide from lotus root

The extraction of polysaccharide from lotus root was highly homogenized, and the structure of the polysaccharide was not clear. Herein, we report a hot water method combined with α-amylase that was applied to extract lotus root polysaccharide. After purified, a lotus root polysaccharide fraction LP60-a with high purity and low molecule weight was obtained. Systematic characterization of the structure of LP60-a was achieved by monosaccharide composition, methylation and NMR analysis, showing that LP60-a was composed of α-1,6-glucan linked with a small amount of arabinogalactan. Conformational determination showed that LP60-a was a three-helix polysaccharide with random coil conformation. Furtherly, the immunomodulatory activities of LP60-a were investigated in RAW264.7 macrophages. The data indicated that LP60-a could enhance the proliferation and phagocytosis of macrophages significantly, and induce the expression of NO and TNF-α in macrophages without causing inflammation. Moreover, LP60-a promoted the phosphorylation of MAPK p38 and JNK, as well as NF-κB p65, indicating that LP60-a could activate RAW264.7 cells through MAPK and NF-κB signaling pathways. In conclusion, the results imply that LP60-a could enhance the immune function of macrophages, presenting a possibility to play a role as an immunomodulatory agent in dietary supplements.

Effects of Platycodon grandiflorus on doxorubicin resistance and epithelial-mesenchymal transition of breast cancer cells via the p38 mitogen-activated protein kinase pathway.

With the increase of chemotherapy frequency for breast cancer, the drug resistance rate of patients is rising, accompanied by cell invasion and metastasis, thus causing mortality. We aimed to explore the mechanism by which Platycodon grandiflorus affects breast cancer cells in terms of the doxorubicin (Dox) resistance and epithelial-mesenchymal transition (EMT) via the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. MCF-7/R cell lines with resistance to Dox were established. After 24h of culture with DMEM (blank group), they were divided into Platycodon grandiflorus, Platycodon grandiflorus + Ophiopogon japonicus, Platycodon grandiflorus + Curcumae Rhizoma, Platycodon grandiflorus + Curcumae Rhizoma + U46619 groups. Flow cytometry, colony formation assay, as well as Transwell assay were performed to examine the cells for apoptosis, proliferation, and invasion, respectively. Western blotting was performed to measure the phosphorylated (p)-p38 MAPK-to-p38 MAPK ratio together with N-cadherin, vimentin, β-catenin, and E-cadherin protein expressions. Compared with the blank group, the half maximal inhibitory concentration (IC50), number of cell colonies, number of invading cells and expressions of proteins related to EMT (i.e. N-cadherin, vimentin, and β-catenin) significantly reduced, but increases in apoptosis rate, p-p38 MAPK/p38 MAPK ratio and E-cadherin protein expression were observed in different groups (P < 0.05). Compared with the Platycodon grandiflorus + Curcumae Rhizoma group, the Platycodon grandiflorus + Curcumae Rhizoma + U46619 group had significantly decreased IC50, cell colony count, invading cell count and β-catenin, N-cadherin, and vimentin expressions, in addition to elevated E-cadherin protein expression, apoptosis rate, and p-p38 MAPK/p38 MAPK ratio (P < 0.05). Platycodon grandiflorus can reverse the resistance of breast cancer cells to Dox and regulate their biological activities by activating the p38 MAPK signaling pathway.

Downregulation of microRNA-29b in cancer and fibrosis: molecular insights and clinical implications

MicroRNA-29b (miR-29b) is known for its therapeutic potential as an antifibrotic and anticancer agent. In fibrotic conditions, miR-29b inhibits fibrogenesis by downregulating crucial regulators such as collagens, extracellular matrix proteins and the transforming growth factor-β pathway. Similarly, in cancer, it acts as a tumor suppressor by downregulating various oncogenes and signaling pathways involved in cancer progression, such as Wnt-β-catenin, p38-mitogen-activated protein kinase and nuclear factor-κB. However, the upregulation of these pathways suppresses miR-29b, contributing to fibrosis and cancer development. Preclinical research and clinical trials have shown that delivering exogenous miR-29b mimics can restore its expression, attenuating tumorigenesis and fibrogenesis. This review discusses miR-29b's potential and its possible therapeutic development for cancer and fibrotic disorders.

MIR4435-2HG: A novel biomarker for triple-negative breast cancer diagnosis and prognosis, activating cancer-associated fibroblasts and driving tumor invasion through EMT associated with JNK/c-Jun and p38 MAPK signaling pathway activation

BackgroundBreast cancer has the highest incidence rate and causes the most fatalities among all female cancers worldwide. Triple-negative breast cancer (TNBC) is known for its strong invasiveness and higher rates of recurrence. In this research, we aimed to identify MIR4435-2HG as a promising long non-coding RNA (lncRNA) biomarker and therapeutic target for TNBC. MethodsUtilizing clinicopathological information and transcriptome data from The Cancer Genome Atlas (TCGA) database, we assessed the clinical relevance of MIR4435-2HG in breast cancer through univariate and multivariate COX regression, receiver operating characteristic (ROC) analysis, as well as Kaplan-Meier survival analysis. To investigate the biological role of MIR4435-2HG in TNBC, we conducted gene set enrichment analysis (GSEA), as well as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Additionally, we constructed and validated a nomogram to predict disease-free survival (DFS). Both the R package “pRRophetic” and the Tumor Immune Dysfunction and Exclusion (TIDE) algorithm were employed to forecast the sensitivity to different therapeutics between the high- and low-MIR4435-2HG groups. We employed single-cell RNA sequencing analysis and tumor microenvironment infiltration analysis to investigate the potential involvement of MIR4435-2HG in the TNBC tumor microenvironment. Cellular biological behaviors were assessed utilizing CCK-8, transwell assays, and wound-healing assays. Furthermore, we performed RNA-seq, qRT-PCR, and western blotting analyses to elucidate and confirm the specific mechanisms underlying the role of MIR4435-2HG in TNBC. ResultsIn our study, we have identified MIR4435-2HG as a significant diagnostic and prognostic factor for TNBC. We observed that MIR4435-2HG is widely expressed and might have a significant impact on the reshaping of the TNBC tumor microenvironment. Patients with TNBC in the high-MIR4435-2HG group may show reduced sensitivity to cisplatin, doxorubicin, and gemcitabine and have an increased propensity for immune escape. Knockdown of MIR4435-2HG inhibits cancer-associated fibroblasts (CAFs) activation. Notably, MIR4435-2HG predominantly enhances the migratory and invasive capabilities of TNBC cells through the epithelial-mesenchymal transition (EMT) process. Mechanistically, we validated that MIR4435-2HG activates the JNK/c-Jun and p38 non-classical MAPK signaling pathway in TNBC cells. ConclusionsOur findings highlight the significant potential of MIR4435-2HG as a highly promising biomarker for TNBC. Targeting MIR4435-2HG could represent an appealing therapeutic approach for TNBC.

Capsular polysaccharide from the marine bacterium Cobetia marina induces apoptosis via both caspase-dependent and mitochondria-mediated pathways in HL-60 cells

In the present study, we investigated the antiproliferative effect of the capsular polysaccharide (CPS) from marine Gram-negative bacterium Cobetia marina (formerly C. pacifica) KMM 3878 against human leukemia cells in vitro and the potential molecular mechanism underlying this activity. Our results showed that the CPS could inhibit the proliferation of HL-60 cells in a dose-dependent manner with no effect on normal PBMC cells. HL-60 cells treated with the CPS exhibited typical morphologic and biochemical signs of apoptosis. We found that the CPS caused the collapse of mitochondrial transmembrane potential (ΔΨm), activated caspases-8,-9, and − 3, decreased the ratio of anti-apoptotic Bcl-2 and pro-apoptotic Bax proteins, increased ROS production and TNF-α secretion, and stimulated phosphorylation of p38 MAPK and p53 in HL-60 cells. Taken together, these data suggest that both extracellular and intracellular signaling pathways contribute to the CPS-induced apoptosis in HL-60 cells.