TGF-β1 mRNA Expression Research Articles

Expression and correlation analysis of VISTA in peripheral blood mononuclear cells of myasthenia gravis patients.

Myasthenia gravis (MG) is a T cell-dependent, B cell-mediated disorder strongly associated with antigen presentation by dendritic cells (DCs). In MG, mucosal tolerance is linked to increased expression of TGF-β mRNA in monocytes. Additionally, monocytic myeloid-derived suppressor cells (M-MDSCs) exhibit negative immunomodulatory effects by suppressing autoreactive T and B cells. In short, all these cells play an important role in the occurrence of MG. V domain-containing Ig suppressor of T-cell activation (VISTA) is an immune checkpoint molecule constitutively expressed on dendritic cells (DCs), CD4+, CD8+ T cells, monocytes and M-MDSCs. Similar to CTLA-4 and PD-1, VISTA controls peripheral tolerance and autoimmune disorder. VISTA expressed on APCs acts as a ligand to suppress the proliferation and cytokine production of both CD4+ and CD8+ T cells. VISTA expressed on CD4+ T cells also suppresses T cell activation in a T-cell autonomous manner. Thus, VISTA may also play an important role in the immune regulation of MG disease. To investigate the role of VISTA in MG pathogenesis, we collected peripheral blood mononuclear cells (PBMCs) from MG patients and detected VISTA expression in different immune cell populations by FACs. VISTA expression was increased in CD4+ T cells, CD8+ T cells, B cells, monocytes, DCs, and M-MDSCs of PBMCs from MG patients. Correlation analysis further demonstrated that VISTA expression in monocytes was correlated with anti-AChR-IgG levels and MG-ADL scores. VISTA expression was positively associated with IL-4, TGF-β, Foxp3, IL-10, and TNF-α mRNA and negatively associated with IL-1β and IL-6 mRNA in MG PBMCs. VISTA can inhibit expression of IL-6 and TNF-α in vitro. These results suggest that VISTA may modulate MG disease progression and indicate the potential utility of VISTA agonists in MG treatment.

P0191 The role of myeloid mineralocorticoid receptor and its target neutrophil gelatinase-associated lipocalin in mice with colitis

Abstract Background Intestinal fibrosis is a common complication in IBD patients with no specific treatment. In extra-intestinal conditions, mineralocorticoid receptor (MR) antagonism inhibits inflammation and fibrosis in different pathophysiological conditions. We previously showed that pharmacological antagonism of MR also inhibits experimental intestinal fibrosis through its target neutrophil gelatinase-associated lipocalin (NGAL). We now hypothesize that myeloid MR may promote the transition from inflammation to intestinal fibrosis by modulation of macrophage plasticity. We aimed to investigate whether myeloid MR or myeloid NGAL deletion prevents colitis development in mice. Methods Acute and chronic colitis were respectively induced by 1% dextran sodium sulfate (DSS) in myeloid MR knock-out (MyMR KO n=10, WT n= 5) and myeloid NGAL knock-out (MyNGAL KO n=6, WT n=8). Colon tissues were collected for quantitative PCR analysis of inflammatory (Il1, Il6, Arg1, Nos2, Retnla, Adgre1) and fibrotic markers (Col1a1, Tgfb1, Fn1, Mmp3, Mmp9, Igf1, Tnf, Timp1). Fecal calprotectin and lipocalin levels were analyzed by ELISA. Results Myeloid MR deletion significantly reduced colon TGFβ mRNA expression (P<0.05) but did not impact weight loss or intestinal permeability in mice. Myeloid NGAL deletion significantly reduced colitis-induced weight loss (P<0.05) and inhibited fibrosis markers such as collagen 1 and metalloproteinase mRNA expression in mice with chronic colitis (Figure 1). Myeloid MR or NGAL invalidation did not significantly affect fecal calprotectin and lipocalin levels in our experimental conditions. Conclusion In our pilot study, myeloid MR and myeloid NGAL deletion inhibits fibrosis development but further studies are required to confirm these findings.

New insight into the protective effect of Citrullus colocynthis loaded with ZnONP cream on glutaraldehyde-induced dermatitis in health care workers

Glutaraldehyde (GLU) is mainly used in medicine by healthcare workers during infection control as a chemical disinfectant. It has been linked to numerous health hazards that range from asthma to irritation of the eye to contact dermatitis. Citrullus colocynthis (C.C.) is utilized as a supplement to combat a range of health-related problems. This study aimed to assess the effectiveness of locally applied Citrullus colocynthis extract and Citrullus colocynthis loaded with ZnONPs against dermatitis caused by the disinfectant glutaraldehyde (2%).The female mice were divided into five groups (G1, G2, G3, G4, and G5). Group 1 was used as a control. The other 4 groups (2,3,4,and 5) were sprayed with 2% GLU (2 mg/kg body weight), and the other groups (3,4,and 5) were subjected to local application of natural products (Citrullus colocynthis extract cream, ZnONP cream, and Citrullus colocynthis loaded on ZnONP cream), respectively. Each experimental animal was followed for 5 days per week for 30 days.Our findings revealed that GLU-induced dermatitis via the upregulation of TNF-α, IL-1b, NFkb 1, and ptgs2 mRNA expression and the downregulation of TGFB1 mRNA expression caused oxidative stress and altered the biochemical markers and histological appearance. However, these effects were improved by the ZnONPs, C.C. extract, and C.C.-ZnONPs.Local application of Citrullus colocynthis ZnONPs and ZnONPs had preventive effects against GLU-induced dermatitis through the suppression of oxidative stress and inflammatory markers and the enhancement of antioxidants.

Dietary γ-Aminobutyric Acid Promotes Growth and Immune System Performance and Improves Erythropoiesis and Angiogenesis in Gibel Carp (Carassius auratus gibelio).

This experiment aimed to investigate the effect of dietary supplementation of γ-aminobutyric acid (GABA) on the growth performance, immune response, and oxygen-transport-related factors of Gibel carp (Carassius auratus gibelio). An eight-week culturing experiment was designed with five experimental diets, with the actual GABA content being 368 mg/kg (G1, control group), 449 mg/kg (G2), 527 mg/kg (G3), 602 mg/kg (G4), and 675 mg/kg (G5). The results showed that the level of 527 mg/kg (G3) of GABA significantly increased the specific growth rate (SGR), weight gain rate (WGR), and final body weight (FBW) of Gibel carp, while the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), and glucose (GLU) were also increased significantly. In addition, 527 mg/kg (G3) and 602 mg/kg (G4) of GABA significantly increased the total antioxidant capacity (T-AOC). The mRNA expression of tnf-α, tgf-β, and il-10 was significantly increased at the level of 449 mg/kg (G2). In terms of oxygen-carrying capacity, the mRNA expression of epo, tf, tfr1, ho-1, and vegf was markedly increased at the level of 449 mg/kg (G2). In conclusion, dietary GABA supplementation can boost growth performance, enhance the immune system, and increase oxygen-carrying capacity in Gibel carp.

Mechanical Strain of Corneal Epithelium Influences the Expression of Genes Implicated in Keratoconus.

Although mechanical injury to the cornea (e.g. chronic eye rubbing) is a known risk factor for keratoconus progression, how it contributes to loss of corneal integrity is not known. Here, we set out to determine how eye rubbing can influence keratoconus progression by exploring the expression of known disease markers in mechanically stressed corneal epithelial cells. To explore the effects of mechanical stress on the expression of genes implicated in keratoconus (e.g. WNT10A, COL12A1, and TGFB1), we measured their expression using an in vitro model that simulates eye rubbing by cyclic stretching of an immortalized human corneal epithelial cell line (hTCEpi) for 16hours. We further examined the influence of WNT10A expression in hTCEpi cells using loss-of-function approaches. Mechanical strain led to a marked reduction in WNT10A mRNA and protein expression, as well as decreased collagen XII mRNA and protein expression, in hTCEpi cells. Reduced expression of WNT10A protein in WNT10A knockdown cells resulted in reduced protein expression of collagens I and XII, and reduced mRNA expression of MMP9 and TGFB1. Conversely, primary keratocytes treated with recombinant WNT10A protein increased TGFB1 mRNA expression. We provide a molecular explanation for how mechanical strain results in reduced expression of WNT10A in the corneal epithelium, which, in turn, leads to depletion of collagen type I and XII, and TGFβ1 expression. These results provide a molecular link among mechanical strain, WNT10A expression, and the biomechanical failure of the keratoconus cornea.

Evaluating the role of active TGF-β1 as inflammatory biomarker in Kashmiri (North-Indian) patients with systemic sclerosis: a case-control study

Abstract Background As a master immune system regulator, transforming growth factor β1 (TGF-β1) is closely linked to the complicated pathophysiology and development of systemic sclerosis (SSc), a multisystem fibrotic disease. Objective We aim to evaluate the transcriptional levels of TGF-β1 mRNA in PBMCs, assess the TGF-β1 serum levels of SSc patients, and compare them with those of healthy subjects. Methods PBMCs were isolated from whole blood of 50 SSc patients and in 30 healthy controls. After total RNA was extracted from isolated PBMCs, complementary DNA (cDNA) synthesis was performed. Afterward, the expression of TGF-β1 mRNA was assessed using quantitative real-time PCR using the SYBR Green, GAPDH, and TGF-β1 specific primers. The serum levels of TGF-β1 were determined using a commercially available ELISA kit. Results There was a significant upregulation of TGF-β1 relative expression (p < 0.0001), when SSc patients were compared to the control group. The diffuse subgroup was more common in patients with elevated TGF-β1 mRNA expression (p < 0.0001). However, an insignificant difference was observed between the disease subsets of SSc. Serum TGF- β1 levels were upregulated in SSc patients (78.35 ± 23.16) compared to healthy subjects (61.06 ± 15.90), and were considerably higher in SSc patients with ILD (p < 0.01) and positive anti-topo-Isomerase antibody (p < 0.0001). Conclusion In patients with SSc, elevated levels of TGF-β1 in serum and their correlation with clinical symptoms imply that this cytokine may serve as a marker for fibrotic and vascular involvement in SSc.

Selenium Enhances the Growth of Bovine Endometrial Stromal Cells by PI3K/AKT/GSK-3β and Wnt/β-Catenin Pathways.

The bovine uterus is susceptible to bacterial infections after calving, particularly from Escherichia coli (E. coli), which often results in endometritis. Additionally, postpartum stress in cows can elevate cortisol levels in the body, inhibiting endometrial regeneration and reducing immune function, thereby further increasing the risk of infection. Selenium (Se) is a common feed additive in dairy farming, known for its anti-inflammatory and antioxidant effects. The aim of this study was to investigate the regulatory role of Se in the growth of bovine endometrial stromal cells (BESCs) under the conditions of LPS-induced inflammatory damage at high cortisol levels. BESCs were treated with 1, 2, 4 μM Se in combination with co-treatment of LPS and cortisol. The results indicated that LPS inhibited the cell viability and reduced the mRNA expression of CTGF, TGF-β1, and TGF-β3. Additionally, LPS increased apoptosis, hindered the cell cycle progression by blocking it in the G0/G1 phase, and suppressed the PI3K/AKT/GSK-3β and Wnt/β-catenin signaling pathways. Furthermore, increased concentrations of cortisol can exacerbate the impacts of LPS on cell proliferation and apoptosis. Conversely, the supplementation of Se promoted cell viability, increased the mRNA expression of TGF-β1 and TGF-β3, and enhanced cell cycle progression, while simultaneously repressing cell apoptosis as well as activating the PI3K/AKT/GSK-3β and Wnt/β-catenin signaling pathways. The above findings demonstrated that Se can promote cell proliferation, reduce cell apoptosis, and aid in the growth of BESCs damaged by LPS under high levels of cortisol. The potential mechanisms may be associated with the regulation of the PI3K/AKT/GSK-3β and Wnt/β-catenin signaling pathways.

The effect of tocilizumab treatment for skin fibrosis by inhibiting CD38+ macrophages in systemic sclerosis.

Dermal and pulmonary fibrosis are the main clinical symptoms of systemic scleroderma (SSc), for which there are no effective therapeutic agents. Tocilizumab is thought to improve the symptoms of fibrosis, but the effect of tocilizumab on dermal fibrosis has not been explored. This study aims to investigate the therapeutic effect of tocilizumab on skin fibrosis by inhibiting CD38+ macrophages in the bleomycin-induced SSc mice model. The 8-week-old BALB/c mice were randomly divided into three groups: control group (PBS group), model group (BLM group), and tocilizumab group (TCZ group). The mRNA expression of VIMENTIN, TIMP1, and COL1A1 was measured by qPCR. Western blot was used to detect the protein expression of α-SMA, TGF-β, and COL1A1 in skin tissues. The expression of CD38+ macrophages in the BLM-induced fibrosis mouse model was verified by flow cytometry and immunofluorescence. In comparison to the PBS control group, mice in the BLM group showed skin fibrosis, edema, thickness, and collagen deposition. The percentage of macrophages in the skin, peripheral blood, and spleen was significantly increased in the BLM group, and the percentage of CD38+ macrophages increased in the skin and peripheral blood but decreased in the spleen. After co-cultured with macrophages, L929 fibroblasts differentiated into myofibroblasts, with increased mRNA expression of COL1A1, COL3A, TGF-β, and Fibronectin. Furthermore, after being stimulated by LPS, RAW264.7 cells showed increased expression of IL-6 and CD38. The mRNA levels of COL1A1, COL1A2, COL3A, TGF-β, and Fibronectin in L929 fibroblasts were markedly increased when co-cultured with LPS-stimulated RAW264.7 cells. Tocilizumab treatment reduced dermal thickness and collagen deposition induced by BLM. Furthermore, the percentage of total macrophages and CD38+ macrophages in the skin and peripheral blood significantly decreased after tocilizumab treatment. This study revealed that tocilizumab improved skin fibrosis in the SSc mice model, which was mediated by inhibiting skin and peripheral CD38+ macrophages.

Duhuo Jisheng Decoction in reduction of inflammatory response via Transforming growth factor-β/Smad signaling pathway for repairing rabbit articular cartilage Injury: A Randomized Controlled Trial

ObjectiveThis study aims to investigate the mechanism underlying the effect of Duhuo Jisheng Decoction on the repair of rabbit articular cartilage injury through a reduction in the inflammatory response mediated by the Transforming growth factor (TGF)-β/Smad signaling pathway. MethodsA rabbit articular cartilage injury model was constructed using a ring bone extraction drill. Twenty-four Japanese white rabbits were randomly divided into six groups, namely Sham operation, model, low-dose Duhuo Jisheng Decoction, medium-dose Duhuo Jisheng Decoction, high-dose Duhuo Jisheng Decoction, and positive control groups. The treatment lasted 12 weeks. Gross observation, International Cartilage Repair Society score, Wakitani score, and Micro-computed tomography analysis were used to evaluate the structural repair of cartilage injury. Histology and immunohistochemistry were used to observe the proteoglycan, P-TβRII, P-Smad2, and type II collagen expression levels. Enzyme-linked immunosorbent assay was used to analyze the concentrations of Matrix Metalloproteinase-13 and Syndecan-4 in the joint fluid; and RT-PCR and Western Blot were used to observe the mRNA and protein expressions of ALK5, Sox-9, P-Smad3, and TGF-β1 at the injury repair site. ResultsThe repair effect of cartilage injury, as seen through gross observation and quantitative scoring, was better in all the Duhuo Jisheng Decoction treatment groups than in the model group. The medium dose group of Duhuo Jisheng Decoction had the best repair effect. We observed remarkable structural restoration of cartilage injury in the medium-dose Duhuo Jisheng Decoction group, with the subchondral bone presenting a distinct hierarchy, and parameters such as bone volume fraction and trabecular separation/spacing being significantly augmented. We found high expression levels of proteoglycans, P-TβRII, P-Smad2, and type II collagen. The concentrations of Matrix Metalloproteinase-13 and Syndecan-4 in the joint fluid were significantly lower following treatment. The low gene expression levels of ALK5, Sox-9, P-Smad3, and TGF-β1 in the injury site of the model group could be reversed in the medium-dose Duhuo Jisheng Decoction group. ConclusionDuhuo Jisheng Decoction can repair rabbit cartilage injury and reverse the levels of inflammatory factors in the joint fluid. The mechanism underlying its therapeutic effect is related to the activation of the TGF-β/Smad signaling pathway. This study provides a reliable basis for using Duhuo Jisheng Decoction to treat cartilage injury following knee osteoarthritis.

The potential of supplementing compound organic trace elements at lower levels in Chinese yellow-feathered broiler diets, Part I: Impacts on plasma biochemical parameters, antioxidant capacity, carcass traits, meat quality, and tissue mineral deposition

This study was conducted to evaluate the effects of replacing inorganic trace minerals (ITM) with compound organic trace minerals (OTM) at lower levels on plasma biochemical parameters, antioxidant capacity, carcass traits, meat quality, and tissue mineral deposition in Chinese yellow-feathered broilers. A total of 960 one-day-old male broilers were randomly allocated to six treatment groups. The birds were fed with either the basal diets (negative control, NC), or diets supplemented with 1,000 mg/kg (positive control, PC), 300 mg/kg, and 500 mg/kg ITM or OTM for 53 d, respectively. The results showed that the alkaline phosphatase (ALP) activity of the OTM300 group was significantly higher than that of the NC, PC, and ITM300 groups (P < 0.05). Dietary OTM supplementation could significantly increase the serum concentrations of Fe and Cu, promote the deposition of Zn and Cu in breast muscle, and increase Zn content in the tibia of Chinese yellow-feathered broilers (P < 0.05). Furthermore, dietary OTM300 treatment could significantly increase plasma CAT and CuZn-SOD activities, as well as the CAT activity in the liver (P < 0.05). The liver GSH-Px activity of the OTM500 group were significantly higher than the other groups (P < 0.05). Moreover, the supplementation of dietary OTM could significantly increase the pH45min of breast muscle, as well as decrease drip loss24h and drip loss48h of Chinese yellow-feathered broilers (P < 0.05). Furthermore, pH45min was positively correlated with liver T-AOC activity and the concentrations of Zn, Fe, Cu, and Mn in breast muscle, while drip loss48h was negatively correlated with liver T-AOC activity, plasma CAT and CuZn-SOD, as well as the concentration of Cu and Zn in breast muscle. Trace mineral sources or levels had no significant effect on the carcass traits of Chinese yellow-feathered broilers (P > 0.05). Compared with the ITM groups, OTM300 significantly increased the heart index of Chinese yellow-feathered broilers (P < 0.05). Dietary OTM upregulated the mRNA expression of TGF-β and downregulated the mRNA expression of IL-1β in the spleen (P < 0.05). In conclusion, dietary supplementation with compound OTM at lower levels could promote the deposition of trace minerals in serum and tissues, enhance antioxidant capacity, and improve the meat quality of Chinese yellow-feathered broilers.

FOSL2 activates TGF-β1-mediated GLUT1/mTOR signaling to promote diabetic kidney disease.

Diabetic kidney disease (DKD) is a major cause of kidney failure. FOS-like antigen 2 (FOSL2) has been revealed to be increased in kidney biopsies of patients with lupus nephritis, while its association with DKD remains unsolved. This study aimed to characterize the role of FOSL2 in DKD and its mechanism. The kidney tissues of DKD mice induced by STZ and a high-fat diet were subjected to PAS and Masson's staining. Glomerular mesangial cells (MCs) were treated with high glucose (HG) or normal glucose (NG). CCK-8 and EdU assays were performed to detect cell proliferation, and immunoblotting was conducted to analyze ECM deposition. ChIP-qPCR was performed on MCs to detect the binding of FOSL2 on the TGF-β1 promoter and a dual-luciferase assay to detect the impact of FOSL2 on the transcription of the TGF-β1 promoter. FOSL2 was elevated in the kidney tissues of DKD mice. Knockdown of FOSL2 reduced the mRNA expression of TGF-β1 to decrease the protein expression of GLUT1 and mTOR in the kidney tissues of DKD mice, and TGF-β1 reversed the effects caused by knockdown of FOSL2. The mTOR inhibitor Rapamycin alleviated kidney injury in the presence of FOSL2. Knockdown of FOSL2 inhibited the proliferation and improved ECM deposition of MCs, which were reversed by TGF-β1. Rapamycin and GLUT1 inhibitor BAY-876 reversed the promotion effect of FOSL2 on the proliferation of NG-MCs/HG-MCs and improved ECM deposition of MCs. Our data demonstrated that FOSL2 accentuates DKD in mice by increasing TGF-β1-induced GLUT1/mTOR signaling.

Effects of Dietary Cobalt Levels on Growth Performance, Antioxidant Capacity, and Immune Status of Juvenile Largemouth Bass (Micropterus salmoides).

A 9-week experiment investigated the effects of dietary cobalt levels on the growth performance, antioxidant capacity, and immunity of largemouth bass. Six feed groups were designed and each group received different cobalt levels, including 0.129 mg/kg (control group), 0.192 mg/kg, 0.201 mg/kg, 0.233 mg/kg, 0.277 mg/kg, and 0.316 mg/kg. The results show that the control group (0.129 mg/kg diet) had the lowest final body weight (FBW), weight gain rate (WGR), and specific growth rate (SGR), and the highest feed conversion ratio (FCR), when compared to the cobalt supplementation groups. Dietary cobalt levels of 0.192 mg/kg increased the body protein content and decreased the body moisture content. Regarding antioxidant capacity, the highest catalase (CAT) activity was found in the 0.277 mg/kg dietary cobalt group, while the malondialdehyde (MDA) content was significantly diminished; the total antioxidant capacity (T-AOC) content and glutathione peroxidase (GSH-Px) activity exhibited the highest values in the 0.192 mg/kg and 0.233 mg/kg dietary cobalt groups, respectively. Regarding gene expression, compared with the control group, the mRNA expression of sod was upregulated in the 0.192 mg/kg, 0.233 mg/kg, and 0.277 mg/kg dietary cobalt groups, while the mRNA expression of gpx was diminished when dietary cobalt levels were below 0.233 mg/kg. In addition, the highest il-10 and tgf-β mRNA expression levels were observed in the 0.201 mg/kg and 0.233 mg/kg dietary cobalt groups, respectively. According to the quadratic regression analysis based on the SGR and FCR, the optimal requirement was 0.24 and 0.26 mg/kg of dietary cobalt, respectively.

TMIC-16. SPECIFIC TARGETING OF THE MRNA EXPRESSION OF TGFB2 LIGAND TO POTENTIALLY ENHANCE IMMUNE THERAPIES IN LOW-GRADE GLIOMAS

Abstract Low-grade glioma (LGG) tumors are characterized by a low infiltration of immune cells, requiring therapies augmenting the immune response in the tumor microenvironment (TME). The Transforming Growth Factor beta (TGFB) pathway contributes to an immunosuppressive environment. We propose targeting the specific inhibition of Transforming Growth Factor Beta 2, TGFB2 mRNA, as an attractive therapeutic strategy using antisense oligonucleotides. We conducted a bioinformatic-led investigation highlighting the ratio of TGFB2:TGFB1 mRNA expression levels in the cohort of 513 TCGA patients, revealing the TGFB2 ratio as a negative prognostic indicator through utilizing a multivariate Cox proportional hazards model that included the TGFB2 ratio and age at diagnosis as linear covariates to predict overall survival (OS). The model showed a 1.8-fold higher TGFB2 ratio relative to TGFB1 mRNA expression correlated to worse OS. Furthermore, the impact of the TGFB2 ratio was significantly affected by the age at diagnosis, in which older patients exhibited worse OS at increased TGFB2 ratios. Increasing the ratio of TGFB2 relative to TGFB1, TGFB3, TGFBR1, or TGFBR2 in LGG patients led to significant increases in multivariate Cox model determined hazards ratios, HR (HR (95% CI range)) of 1.372 (1.224-1.539), 1.434 (1.288-1.560), 1.327 (1.195-1.473), and 1.381 (1.233-1.547) respectively (P&amp;lt;0.0001 for all comparisons). Correlation of TGFB2:TGFB1/TGFB3/TGFBR1/TGFBR2 ratio across 1330 Reactome pathways using Gene Set Enrichment Analysis showed the most positively correlated genes were members of TME pathways including PD-1 signaling, Syndecan interactions, Non-integrin membrane-ECM interactions, EPH-Ephrin signaling, and Collagen biosynthesis and modifying enzymes. These results suggest that the specific targeting TGFB2 in LGG can remodel the TME for immune therapies to improve OS outcomes.

Administration of a combination of COX-2/TGF-β1 siRNAs induces hypertrophic scar fibroblast apoptosis through a TP53 mediated caspase pathway

Hypertrophic scar (HTS) formation is a pathological fibrotic skin disease, with no satisfactory treatments available currently. Inducing apoptosis of HTS-derived fibroblasts (HSFs) are becoming promising approaches. In this research, we aim to improve the technology with co-delivery COX-2 and TGF-β1 siRNAs and further investigate the underlying mechanism. Firstly, the HSFs were transfected with 1 µg/ml COX-2 and/or TGF-β1 siRNAs, and proved that the apoptosis of HSFs was greater induced by COX-2/TGF-β1 siRNAs than either COX-2 or TGF-β1 siRNA alone by flow cytometry. To investigate the impact of co-silencing TGF-β1 and COX-2 mRNA expression in vivo, we established HTSs model in rat tails. Our results confirmed that co-silencing of TGF-β1 and COX-2 mRNA expression could significantly alleviate the HTS formation in vivo. Furthermore, we explored the potential molecular mechanism and revealed that the protein levels of TP53, Bcl-2 and Caspase-3 were downregulated while Bax and Cleaved Caspase-3 were upregulated in the COX-2/TGF-β1 siRNA groups compared with HKP group. Taken together, our results demonstrated that simultaneous silencing of COX-2 and TGF-β1 expression by siRNAs induced HSF apoptosis through a TP53 mediated caspase pathway. Therefore, COX-2/TGF-β1 siRNAs might serve as a novel and effective therapeutic alternative for HTSs treatments.

Modulation of Plet1 expression by N-Acetylglucosamine through the IL-17A-MAPK pathway in an imiquimod-induced psoriasis mouse model.

Psoriasis (Ps) is a chronic inflammatory disorder marked by skin plaque formation, driven by immune dysregulation and genetic factors. Despite the available treatments, incidence of Ps is increasing in the dermatology patients. Novel strategies are crucial due to current treatment limitations. The interleukin 17 (IL-17) pathway is pivotal in Ps pathogenesis, however the expression of its putative target gene placenta expressed transcript 1 (Plet1) remains unstudied in Ps. Considering the potential anti-inflammatory properties of N-Acetylglucosamine (GlcNAc), our study explored its role in modulating Plet1 expression in an imiquimod (IMQ)-induced Ps mouse model. Our data demonstarted a significant reduction of inflammation and Psoriasis Area and Severity Index (PASI) scores, downregulation of growth factors (GFs), IL-17A, and MAPK expression after GlcNAc treatment. In addition, GlcNAc treatment reduced neutrophils, monocyte-dendritic cells (Mo-DC) and conventional T cells (Tcons) while increasing monocyte-macrophages (Mo-Macs) and regulatory T cells (Tregs). GlcNAc treatment also downregulated Plet1 overexpression in psoriatic mouse skin and in vitro, reduced proliferation and apoptosis in IL-17A stimulated human dermal fibroblasts (HDF), along with IL-17A and TGF-β mRNA expression. Together, these data suggest that, GlcNAc interferes with downstream mechanisms in IL-17 pathway and downregulating Plet1 expression, presenting a promising strategy for Ps treatment.

Structural characterization, antioxidant and immunomodulatory activities of a polysaccharide from a traditional Chinese rice wine, Guangdong Hakka Huangjiu

Hakka Huangjiu, a traditional Chinese rice wine, boasts a rich history and is known for its immunomodulatory, antibacterial, anti-aging and anti-fatigue effects. However, there is limited research on the primary active components and molecular mechanism of the bioactivity of Hakka Huangjiu. To address this gap, this study assessed the structural characteristics, antioxidant, and immunomodulatory activities of the polysaccharide-1 of Guangdong Hakka Huangjiu (HP1). Structural analysis revealed that HP1 had a low molecular weight polysaccharide of 5550 Da, primarily consisting of glucose (93.2 %), with smaller amounts of xylose, mannuronic acid and galactose. Methylation and NMR analysis suggested that the main glycosidic linkages present in HP1 are α-D-Glcp-(1→, →4)-α-D-Glcp-(1 → and →6) -α-D-Glcp-(1→. Furthermore, HP1 exhibited dose-dependent DPPH·, ABTS+ and OH· scavenging activity. HP1 exhibited significant protection of HepG2 cells from H2O2 damage. Additionally, HP1 induced the release of NO, TNF-α, IL-6 and iNOS in RAW264.7 cells. HP1 treatment significantly increased mRNA expression of TNF-α, IL-6, iNOS, COX-2, IL-1β and TGF-β1. These results suggested that polysaccharides HP1 may have potential as a novel natural antioxidant and immunomodulatory product for use in nutraceuticals and functional foods.

The Role of Dicrocoelium dendriticum Egg Antigen in Colitis: A Molecular, Pathological and Serological Study in an Experimental Model of C57BL/6 Mice.

Inflammatory bowel disease (IBD) is a chronic and recurrent disease of the gastrointestinal tract that enhances the chance of developing colorectal cancer. Since standard treatments such as Mesalazine have limited effectiveness and are often accompanied by numerous side effects, the use of immune modulators derived from worms has been proposed as a new immunotherapy method for inflammatory diseases such as ulcerative colitis. The aim of this study is to investigate the protective effects of D. dendriticum egg antigen on DSS-induced colitis in C57BL/6 mice. D. dendriticum egg antigen was extracted and DSS (3.5%) was used to induce colitis in mice. Treatment and prophylaxis included intraperitoneal injections of D. dendriticum egg antigen. Histopathological indicators and the disease activity index (DAI), including weight loss, rectal bleeding, stool consistency, and rectal prolapse, were used to assess the severity of colitis. Real-time PCR measured the expression of transforming growth factor-β (TGF-β) and interleukin-17 (IL-17), while ELISA determined the concentration of these cytokines. Treatment with D. dendriticum egg antigen significantly improved the clinical symptoms and decreased the severity of DSS-induced colitis. Furthermore, D. dendriticum egg antigen increased the expression of TGF-β mRNA and reduced the expression of IL-17 mRNA, leading to a positive adjustment in the regulation of proteins and reduction of inflammatory proteins. As a result, the macroscopic, microscopic inflammation and activity index (DAI) of DSS-induced decreased. D. dendriticum egg antigen provides a promising new way to modulate the immune system and improve ulcerative colitis.

Effect of Buzhong Yiqi Decoction on Treg cells in rats with autoimmune thyroiditis through TGF-β/Smad signaling pathway

To investigate the effect of Buzhong Yiqi Decoction on regulatory T cells(Treg) in experimental rats with autoimmune thyroiditis(EAT) through the transforming growth factor-β(TGF-β)/Smad signaling pathway. Female SD rats were immunized with iodine-rich drinking water combined with Freund's adjuvant and porcine thyroglobulin(pTG) to establish the EAT model of rats, and the levels of serum thyroperoxidase antibody(TPOAb) and thyroglobulin antibody(TGAb) were detected. Pathological sections by hematoxylin-eosin(HE) staining were observed. Treg in the rats' spleen were extracted by immunomagnetic beads after the successful modeling and identified by flow cytometry. The extracted Treg were divided into blank group, Buzhong Yiqi Decoction group, TGF-β group, antagonist(LY3200882), and antagonist(LY3200882)+Buzhong Yiqi Decoction group. After the intervention, the cell counting kit-8(CCK-8) experiment was conducted to detect cell viability. Western blot and quantitative real-time PCR(RT-qPCR) were used to detect the expression of TGF-β/Smad signaling pathway-related proteins and genes. The results showed that the levels of TPOAb and TGAb increased in the rats in the model group compared to the rats in the blank group. HE staining showed that part of the follicles in the thyroid tissue of the rats in the model group were destroyed, and a large number of lymphocytes were infiltrated, indicating that the modeling was successful. After Treg were administered in vitro, CCK-8 results showed that the serum concentration of Buzhong Yiqi Decoction was below 40% to promote cell proliferation. The Buzhong Yiqi Decoction-containing serum group could increase the protein expression of TGF-β1, FoxP3, Smad2, and Smad4 compared with the blank serum group, while the expression of p-Smad2, p-Smad3, and Smad3 increased compared with the blank serum group, but the difference was not statistically significant. Compared with the antagonist group, the protein expressions of p-Smad2, Smad2, p-Smad3, Smad3, Smad4, and Smad7 did not significantly increase or decrease in the antagonist group after adding Buzhong Yiqi Decoction-containing serum. RT-qPCR showed that compared with the blank serum group, the mRNA expression of TGF-β1, FoxP3, Smad2, Smad3, Smad4, and Smad7 in the Buzhong Yiqi Decoction group increased or decreased in the same trend as that in the TGF-β group, but there was no statistical significance. After Buzhong Yiqi Decoction-containing serum was added to the antagonist group, the mRNA levels of TGF-β1, FoxP3, Smad2, Smad3, Smad4, and Smad7 were not statistically significant. In conclusion, Buzhong Yiqi Decoction could promote the stability and activity of Treg cells by promoting the secretion of TGF-β1 and regulating the expression of key signaling molecules TGF-β1, Smad2, and Smad4 in the TGF-β/Smad signaling pathway, thus affecting the immune balance of Th17/Treg and inhibiting the inflammatory response of rats with EAT.

Polysaccharides from Sacha Inchi shell reduces renal fibrosis in mice by modulating the TGF-β1/Smad pathway and intestinal microbiota

Renal fibrosis is a common pathway involved in the progression of various chronic kidney to end-stage diseases, posing a substantial global public health challenge in the search for effective and safe treatments. This study investigated the effects and mechanisms of sacha inchi shell polysaccharide (SISP) on renal fibrosis induced by a high-salt diet (HSD) in mice. By analysing kidney-related protein pathways and the structure of gut microbiota, we found that SISP significantly reduced urinary protein levels induced by a HSD from 41.08 to 22.95 μg/mL and increased urinary creatinine from 787.43 to 1294.50 μmol/L. It reduced renal interstitial collagen fibres by 11.30 %, thereby improving the kidney function. SISP lowered the mRNA expression of TGF-B1, fibronectin, α-SMA, Smad2/3, and TGFBRII, leading to decreased protein levels of TGF-β1, p-Smad2/3, p-TGFβRII, fibronectin, α-SMA, p-Smad2/3/Smad2/3, and p-TGFβRII/TGFβRII. These changes blocked downstream transcription in the TGF-β1/Smad signalling pathway, thereby attenuating renal fibrosis in HSD mice. In addition, SISP altered the intestinal flora imbalance in HSD mice by reducing the relative abundance of the genera, Akkermansia, Faecalibaculum, and unidentified_Ruminococcaceae, and reversing the decline in the levels of the genera, Lactobacillus and Bacteroides. In conclusion, SISP is a promising nutraceutical for renal fibrosis management.

Gastrodin improves microglia-mediated inflammatory response after hypoxic-ischemic brain damage in neonatal rats via PI3K/AKT pathway

To investigate the mechanism of gastrodin for inhibiting microglia-mediated inflammation after hypoxicischemic brain damage (HIBD) in neonatal rats. Thirty-nine 3-day-old SD rats were randomly divided into sham group, HIBD group and gastrodin treatment group. Western blotting was used to detect the expressions of TNF-α, IL-1β, IL-10 and TGF- β1 in the corpus callosum of the rats. The potential targets of gastrodin for treatment of HIBD were screened by network pharmacology analysis. The expressions of PI3K/AKT signaling pathway proteins following HIBD-induced microglial activation in the rats and in cultured microglial BV-2 cells with oxygen-glucose deprivation (OGD) were detected with Western blotting. The effects of LY294002 (a specific inhibitor of the PI3K/AKT pathway) and gastrodin on TNF-α and TGF-β1 mRNA levels in BV-2 cells with OGD was detected with RT-qPCR. In the neonatal rats with HIBD, gastrodin treatment significantly decreased TNF-α and IL-1β expressions and enhanced IL-10 and TGF-β1 expressions in the ischemic corpus callosum. Network pharmacology analysis showed significant enrichment of the PI3K/AKT signaling pathway and a strong binding between gastrodin and PI3K. Gastrodin significantly promoted PI3K and AKT phosphorylation in neonatal rats with HIBD and in BV-2 cells exposed to OGD. In BV-2 cells with OGD, gastrodin obviously suppressed OGD-induced increase of TNF-α and reduction of TGF-β1 mRNA expressions, and this effect was strongly attenuated by LY294002 treatment. Gastrodin can inhibit microglia-mediated inflammation in neonatal rats with HIBD by regulating the PI3K/AKT signaling pathway.