Increased Expression Of TGF-β1 Research Articles

Strength training attenuates neuropathic pain by Preventing dendritic Spine dysgenesis through Suppressing Rac1 and inflammation in experimental autoimmune encephalomyelitis

Chronic pain is a challenge and major health problem to basic science and clinical practice. Pain is one of the worst symptoms of multiple sclerosis (MS), which has a significant impact on their quality of life. Rac1 is an important intracellular signaling molecule involved in spinal dendritic spine pathology and activation of IL-1β and TNF-α that are associated with chronic neuropathic pain. As a result, targeting Rac1 presents a promising approach to managing neuropathic pain. Clinical studies have demonstrated that physical exercise is a non-pharmacological strategy that positively influences disease progression in individuals with MS, but underlying mechanism of exercise on Rac1- induced neuropathic pain is not well understood. This study examined the effects of a 4-week strength training on Rac1 expression, IL-1B, TNF-α, TGF-β1 levels, MDA concentrations, SOD activity, dendritic spine abnormalities in the dorsal horn of the spinal cord, as well as nociceptive behaviors (formalin test) and motor function (Rotarod test) during the chronic phase of experimental autoimmune encephalomyelitis (EAE). The findings indicated that strength training increased TGF-β1 expression and SOD activity while decreasing the expression of Rac1, IL-1β, TNF-α, and MDA and reducing dendritic spine dysgenesis in the dorsal horn of the spinal cord. We observed strength training effectively reduced nociceptive behaviors and improved motor function in mice with EAE. In summary, regular physical exercise may modulate neuropathic pain through inhibition of dendritic spine dysgenesis, inflammation and oxidative stress in the dorsal horn of the spinal cord.

Abstract 4134967: Pericardial Adipose Tissue Hypertrophy Contributes to Heart Failure via β3 Adrenergic Signaling-Mediated Activation of the TGFβ1 Pathway

Background: The heart is surrounded by pericardial adipose tissue (PeAT). An increase in PeAT volume has been associated with the development of heart failure; however, the precise molecular mechanisms by which hypertrophic PeAT contributes to heart failure remain to be elucidated. Aims: To clarify the involvement of PeAT hypertrophy in the development of heart failure. Methods: Male C57BL/6 mice (8-10 weeks old) were fed a high-fat diet (HFD) for 8 weeks to induce PeAT hypertrophy. Cardiac pressure overload was induced by transverse aortic constriction (TAC) with a 26-gauge needle. Results: PeAT volume significantly increased in HFD-fed mice compared to those on a normal diet (ND) (25.28 ± 4.1 mg vs. 9.35 ± 0.73 mg, p<0.01). Eight weeks after TAC, HFD-fed mice (TAC+HFD) exhibited more pronounced cardiac dysfunction (fractional shortening: 45.21 ± 2.21% vs. 55.99 ± 1.91%, p=0.002, left ventricular mass: 130.3 ± 7.80% vs. 106.8 ± 3.77%, p=0.0154), upregulation of cardiac fibrosis-related genes ( Col1a1 , Col3a1 ), and greater fibrotic area in the left ventricle compared to ND-fed TAC mice (TAC+ND). Simultaneously, the TGFβ/Smad pathway was activated in the left ventricles of TAC+HFD mice. Notably, surgical removal of hypertrophic PeAT improved cardiac function in TAC+HFD mice, while transplantation of PeAT from HFD-fed mice into TAC+ND mice worsened cardiac function. In TAC+HFD mice, local noradrenaline concentration in PeAT decreased, along with reduced activation of hormone-sensitive lipase, a downstream target of β3 adrenergic receptor signaling. Local free fatty acid concentrations also decreased in the PeAT of HFD-fed mice. In the RAW 264.7 macrophage cell line, a decrease in palmitic acid, a major fatty acid, led to increased expression of TGFβ1. Conclusions: These results suggest that HFD-induced PeAT hypertrophy suppresses β3 adrenergic receptor signaling and free fatty acid release, increasing TGFβ1 expression in the macrophages and resulting in cardiac fibrosis and dysfunction. Our findings provide new insights into the pathogenesis of PeAT hypertrophy in heart failure development.

Exploring the Anti-Inflammatory and Antioxidative Potential of Selenium Nanoparticles Biosynthesized by Lactobacillus casei 393 on an Inflamed Caco-2 Cell Line.

Selenium (Se) plays a crucial role in modulating inflammation and oxidative stress within the human system. Biogenic selenium nanoparticles (SeNPs) synthesized by Lactobacillus casei (L. casei) exhibit anti-inflammatory and anti-oxidative properties, positioning them as a promising alternative to traditional supplements characterized by limited bioavailability. With this context in mind, this study investigates the impact of selenium and L. casei in ameliorating inflammation and oxidative stress using a cell line model. The study is centered on the biosynthesis of selenium nanoparticles (SeNPs) by L. casei 393 under anaerobic conditions using a solution of sodium selenite (Na2SeO3) in the bacterial culture medium. The generation of SeNPs ensued from the interaction of L. casei bacteria with selenium ions, a process characterized via transmission electron microscopy (TEM) to confirm the synthesis of SeNPs. To induce inflammation, the human colonic adenocarcinoma cell line, Caco-2 was subjected to interleukin-1 beta (IL-1β) at concentrations of 0.5 and 25 ng/ml. Subsequent analyses encompass the evaluation of SeNPs derived from L. casei, its supernatant, commercial selenium, and L. casei probiotic on Caco2 cell line. Finally, we assessed the inflammatory and oxidative stress markers. The assessment of inflammation involved the quantification of NF-κB and TGF-β gene expression levels, while oxidative stress was evaluated through the measurement of Nrf2, Keap1, NOX1, and SOD2 gene levels. L. casei successfully produced SeNPs, as confirmed by the color change in the culture medium and TEM analysis showing their uniform distribution within the bacteria. In the inflamed Caco-2 cell line, the NF-κB gene was upregulated, but treatment with L. casei-SeNPs and selenium increased TGF-β expression. Moreover, L. casei-SeNPs upregulated SOD2 and Nrf2 genes, while downregulating NOX1, Keap1, and NF-κB genes. These results demonstrated the potential of L. casei-SeNPs for reducing inflammation and managing oxidative stress in the Caco-2 cell line. The study underscores the ability of L. casei-SeNPs to reduce oxidative stress and inflammation in inflamed Caco-2 cell lines, emphasizing the effectiveness of L. casei as a source of selenium. These insights hold significant promise for the development of SeNPs derived from L. casei as potent anti-inflammatory and anti-cancer agents, paving the way for novel therapeutic applications in the field.

Activation of the Alpha 7 Nicotinic Acetylcholine Receptor by GTS-21 Mitigates Contrast Nephropathy in a Rat Model

Introduction: Contrast nephropathy (CN) is characterized by oxidative stress, vasoconstriction, tubular toxicity, and hypoxia of the renal medulla. We aimed to test the therapeutic effects of an α7 nicotinic acetylcholine receptor (nAChR) agonist, GTS-21, in an experimental CN model. Methods: Male Sprague-Dawley rats (n = 40) were divided into 4 groups: saline-treated control, GTS-21-treated control, contrast, and GTS-21-treated contrast groups. Starting on the 1st day, GTS-21 (4 mg/kg, intraperitoneally) or saline was administered twice a day for 3 days. CN was induced on the second day by intravenous injection of indomethacin (10 mg/kg), l-NAME (10 mg/kg), and a contrast agent with high osmolarity (6 mL/kg; Urografin 76%). At the 72nd hour, blood and kidney samples were obtained for the determination of biochemical, histological, and gene expression parameters. Results: Compared to those in control rats, the elevated serum BUN level in the contrast group decreased with GTS-21 treatment, while H&E staining and TUNEL assays showed that contrast-induced renal injury was improved by GTS-21. Moreover, GTS-21 treatment in the CN also increased the antioxidant glutathione level. In the contrast group, a significant increase in IL-6 expression and a decrease in TGF-β expression were observed; however, GTS-21 treatment decreased IL-6 expression and increased TGF-β expression. Conclusion: GTS-21 significantly alleviated renal injury parameters through antioxidant, anti-inflammatory, and antiapoptotic mechanisms in the CN model.

Transforming growth factor-β1 and vascular endothelial growth factor levels in senile acute myeloid leukemia and correlation with prognosis.

Acute myeloid leukemia (AML) is a disease in which immature hematopoietic cells accumulate in the bone marrow and continuously expand, inhibiting hematopoiesis. The treatment and prognosis of this disease have always been unsatisfactory. To investigate the correlation between vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGFβ1) expression and prognosis in older adults with AML. This study enrolled 80 patients with AML (AML group), including 36 with complete response (AML-CR), 23 with partial response (AML-PR), and 21 with no response (AML-NR). The expression levels of VEGF and TGFβ1 were detected by reverse transcription polymerase chain reaction in bone marrow mononuclear cells isolated from 56 healthy controls. Kaplan-Meier analysis was performed to assess overall survival (OS) and progression- or disease-free survival (DFS). Prognostic risk factors were analyzed using a Cox proportional hazards model. The AML group showed a VEGF level of 2.68 ± 0.16. VEGF expression was lower in patients with AML-CR than those with AML-PR or AML-NR (P < 0.05). TGFβ1 expression in the AML group was 0.33 ± 0.05. Patients with AML-CR showed a higher TGFβ1 expression than those with AML-PR or AML-NR (P < 0.05). VEGF and TGFβ1 expression in patients with AML was significantly correlated with the counts of leukocytes, platelets, hemoglobin, and peripheral blood immature cells (P < 0.05); Kaplan-Meier survival analysis revealed that patients with high TGFβ1 expression had better OS and DFS than those with low TGFβ1 expression (P < 0.05), whereas patients with low VEGF levels showed better OS and DFS than those with high VEGF levels (P < 0.05). VEGF, TGFβ1, and platelet count were identified by the Cox proportional hazards model as independent risk factors for OS (P < 0.05), while VEGF, TGFβ1, and white blood cell count were independent risk factors for DFS (P < 0.05). Decreased VEGF expression and increased TGFβ1 expression in patients with AML provide valuable references for determining and individualizing clinical treatment strategies.

The features of endometrial receptivity in women underwent in vitro fertilization procedure

The objective: to determine the endometrial receptivity markers in women who underwent the procedure of in vitro fertilization (IVF). Materials and methods. The study included 191 patients who applied to the reproductive department of Caspian International Hospital from 2020 to 2022 for an examination for infertility.The main group included 50 patients with infertility who underwent endometrial biopsy during the implantation window (7th–9th days after ovulation in the cycle preceding the IVF stage); the control group – 20 women who underwent treatment for male infertility with a positive program result.The patients underwent IVF according to the standard method. The following expression levels were determined: leukemia inhibiting factor (LIF); vascular endothelial growth factor (VEGF); transforming growth factor-β1 (TGF-β1). The percentage expression of area indices was estimated.Results. In the women of the main group absence of embryo transfer was in 5 (10%) cases, no pregnancy – 27 (54%) cases, presence of pregnancy – 23 (46%) cases, cycle disruption – 3 (6%) cases.According to the results of a histological examination in women of the control group, complete secretory transformation of the endometrium was determined in 85% of cases. In the main group complete secretory transformation of the endometrium was detected in 48%, defective secretory transformation – in 42%, morphological signs of endometrial changes corresponding to the average secretory phase of the cycle – in 10% of women.A significant decrease (p&lt;0.001) in the expression of LIF in the histological structures (stroma) of the endometrium was revealed in the main study group compared to the control one. The most pronounced differences were observed in the expression level of TGF-β1 in the stroma (approximately 10-fold; p&lt;0.001).Conclusions. Thus, infertile women who underwent IVF have multidirectional changes in endometrial receptivity markers: decrease in LIF expression in the stroma and epithelium, increase in VEGF expression in the stroma, and a significant increase in TGF-β1 expression in the stroma and epithelium. These changes indicate an abnormal functional state of the endometrium and affect the results of IVF in patients (absence of oocyte transfer and failure to conceive).

The Effect of Snakehead Fish (Channa striata) Extract on Dry Socket Wound Healing: TGF-β1 Expression in Rats Model

Background: Dry socket or alveolar osteitis is a condition characterized by the breakdown of blood clots due to excessive fibrinolysis activity, which disrupts the wound healing process. Consumption of nutrition-rich foods such as snakehead fish (Channa striata) rich in albumin, omega-3 fatty acids, vitamins, minerals, and striatin may accelerate delayed wound healing. This study aimed to analyze the role of snakehead fish extract on the expression of TGF-β1 in the healing of dry socket wounds after tooth extraction in Wistar rats. Methods: This study was an in vivo study with a posttest-only control group design. Forty-five Wistar rats were divided into five groups: snakehead fish extract groups with doses of 1/2, 1, 2, 4 times human dose, and a negative control. The lower left incisor teeth of the rats were extracted, then dry socket was induced with adrenaline using a paper point. On days 3, 7, and 14, TGF-β1 expression was assessed using immunohistochemical staining. Results: This study showed significant difference in TGF-β1 expression on days 3, 7, and 14. TGF-β1 expression was higher in the treatment groups than the control (p&lt;0.05). Conclusion: Administration of snakehead fish extract increased TGF-β1 expression. These findings underline the complex interaction between snakehead fish extract and TGF-β1 expression.

Abstract 3092: Smooth Muscle Cell Tgf-β Receptor Knockout Improves Arteriovenous Fistula Maturation Via Attenuation Of Autophagy

Objectives: Arteriovenous fistulae (AVF) are the preferred vascular access for patients with end-stage renal disease (ESRD) but have high rates of primary failure. Models of AVF using mice with chronic kidney disease (CKD) show diminished maturation and increased failure compared to AVF using wild type mice. Since mice with CKD show increased TGF-β1 expression and activity in the AVF model, and TGF-β1 signaling can alter autophagy, we hypothesized that CKD increases autophagy during AVF maturation, whereas inhibition of the smooth muscle cell (SMC) TGF-β receptor would attenuate autophagy and promote AVF maturation. Methods: An aortocaval fistula or sham procedure was created in mice that had CKD induced by 5/6 nephrectomy (5/6 Nx) or sham nephrectomy (0/6 Nx). Myh11-CreER T2 ; Tgfbr1 fl/fl ; Tgfbr2 fl/fl ; mT/mG mice (TGFβRI II-KO) and control C57BL/6J mice were used. The diameter of the IVC was determined using ultrasound at days 0, 7, 14, and 21. AVF were harvested at day 7 or 21 for histology and immunoreactivity. Deidentified human AVF and control vein specimens were collected for analysis from second stage basilic vein transpositions. Results: In C57BL/6J mice, AVF had significantly decreased diameter (p=0.0051), increased wall thickness (p=0.0002), as well as increased immunoreactivity of collagen-1 (p=0.0163), collagen-3 (p=0.0029), TGF-β1 (p=0.0184) and fibronectin (p=0.00321) in mice with CKD compared to control mice (n=6). SMC showed increased markers of autophagy (LC3B and P62) in mice with CKD compared to control mice (LC3B, p&lt;0.0001; P62, p=0.006; n=6); similarly, autophagy makers were increased in human AVF compared with the control veins (LC3B, p=0.0023; P62, p=0.006; n=6). In SMC-TGFβR II-KO mice, AVF had significantly decreased wall thickness (p&lt;0.0001), increased diameter (p=0.0258), and decreased expression of autophagy markers (LC3B, p=0.0014; P62, p=0.0046; n=6). Conclusion: CKD increases neointimal hyperplasia that is associated with increased autophagy during AVF failure, and diminished TGF-β signaling reduces autophagy and wall thickness as well as increases AVF diameter. Suppression of SMC autophagy might be a novel therapeutic approach to improve AFV function.

Ursolic acid inhibits glioblastoma through suppressing TGFβ-mediated epithelial-mesenchymal transition (EMT) and angiogenesis

Found in many fruits and plants, Ursolic acid (UA), a pentacyclic triterpene that occurs naturally, is recognized for its anti-cancer effects, especially in combating glioblastoma. However, the intricate molecular mechanisms underpinning its anti-tumor actions are still not fully understood, despite the recognition of these effects. By examining the functions of epithelial-mesenchymal transition (EMT) and angiogenesis, crucial for glioblastoma progression, and their regulation through Transforming Growth Factor Beta (TGFβ) – a key marker for glioblastoma, our research aims to fill this knowledge gap. This study explores how ursolic acid can block the progression of glioblastoma by precisely targeting TGFβ-triggered EMT and angiogenesis. The findings show that UA successfully blocks the spread, movement, and invasion of glioblastoma cells. Accompanying this, there is a significant reduction in the expression of TGFβ and crucial EMT indicators like snail and vimentin. Furthermore, UA shows a reduction in angiogenesis that depends on the dosage, highlighted by decreased vascular endothelial growth factor (VEGF) in human umbilical vein endothelial cells (HUVECs). Interestingly, increased TGFβ expression in U87 and U251 glioblastoma cell lines was found to weaken UA's anti-tumor properties, shedding more light on TGFβ′s critical function in glioblastoma's pathology. Supporting these laboratory results, UA also showed considerable inhibition of tumor growth in a glioblastoma xenograft mouse model. Overall, our research emphasizes Ursolic acid's promise as a new treatment for glioblastoma and clarifies its action mechanism, mainly by inhibiting TGFβ signaling and thereby EMT and angiogenesis.

CTEN-induced TGF-β1 expression facilitates EMT and enhances paclitaxel resistance in bladder cancer cells.

To investigate the role of C-terminal tensin-like (CTEN) in mediating chemotherapy resistance via epithelial-mesenchymal transition (EMT) in bladder cancer (BC) cells, through the regulation of transforming growth factor-β1 (TGF-β1) expression. Lentiviral vectors were used to create CTEN overexpression and knockdown constructs, which were then introduced into paclitaxel-resistant BC cell lines. The effects of CTEN manipulation on cell proliferation and drug sensitivity was assessed using the CCK-8 assay, and apoptosis was evaluated by flow cytometry. The expression levels of CTEN, TGF-β1, and EMT markers were quantified by RT-qPCR and Western blot analysis. The interaction between CTEN and TGF-β1 and its effect on TGF-β1 methylation were studied using bisulfite sequencing PCR and co-immunoprecipitation. Overexpression of CTEN in BC cells was associated with decreased paclitaxel efficacy, reduced apoptosis, and elevated levels of TGF-β1 and EMT-related proteins. CTEN was found to bind TGF-β1, inhibiting its methylation and thereby promoting TGF-β1 upregulation. This increase in TGF-β1 expression facilitated the EMT process and enhanced drug resistance in BC cells. The induction of TGF-β1 expression by CTEN promotes EMT and increases chemotherapy resistance in BC cells. Targeting CTEN or the EMT pathway could improve chemosensitivity in treatment-resistant BC, suggesting a novel therapeutic strategy to enhance chemotherapy effectiveness.

Effect of melatonin‐induced exosomes on proregenerative and antifibrotic pathways involved in corneal scarring

Aims/Purpose: Melatonin is a time‐and‐concentration dependent pro and antiinflammatory agent that also regulates cell proliferation, migration, angiogenesis, fibrosis, and extracellular matrix synthesis. This study aims to assess the effect of exosomes obtained from melatonin‐treated human limbal mesenchymal stem cells (hLMSCs) on naive hLMSCs and ascertain their influence on antifibrotic and proregenerative pathways involved in corneal scarring.Methods: hLMSCs were harvested from the eye bank of Eskişehir Osmangazi University Hospital and were subsequently isolated and characterized. We then treated hLMSCs (Mel‐treated hLMSCs) with varying concentrations of melatonin followed by isolation and characterization of the procured exosomes. These exosomes were added to enrich cell culture media of naïve hLMSCs (Mel‐exo‐treated hLMSCs) to examine their antifibrotic and proregenerative effects. The expression of miR155, miR29 Transforming Growth Factor Beta 1 (TGFβ1), TGFβ3, Peroxisome Proliferator‐Activated Receptor Gamma (PPARγ), and Alpha‐Smooth Muscle Actin (α‐SMA) miRNA and genes were compared between Mel‐treated hLMSCs and Mel‐exo‐treated hLMSCs by using Real‐Time PCR.Results: TGFβ3 expression increased dramatically in response to Mel‐exo, while the increase in TGFβ1 expression was only minimal in correlation with levels of miR155 expression. Likewise, PPARγ expression was increased while an inhibition was observed in α‐SMA expression and miR29 expression in Mel‐exo‐treated hLMSCs compared to Mel‐treated hLMSCs.Conclusions: Melatonin treatment of hLMSCs resulted in expression of exosomes that could drive naïve hLMSCs in favour of antifibrotic and proregenerative effects as evidenced by higher TGFβ3 and PPARγ expressions and inhibition of α‐SMA. This approach could prove beneficial in cornea and ocular surface tissue engineering applications.The authors acknowledge the support from Eskisehir Osmangazi University, Scientific Research Projects (Grant ID: TOA‐2022‐2458).

Chk2 Modulates Bmi1-Deficiency-Induced Renal Aging and Fibrosis via Oxidative Stress, DNA Damage, and p53/TGFβ1-Induced Epithelial-Mesenchymal Transition.

Renal aging may lead to fibrosis and dysfunction, yet underlying mechanisms remain unclear. We explored whether deficiency of the Polycomb protein Bmi1 causes renal aging via DNA damage response (DDR) activation, inducing renal tubular epithelial cell (RTEC) senescence and epithelial-mesenchymal transition (EMT). Bmi1 knockout mice exhibited oxidative stress, DDR activation, RTEC senescence, senescence-associated secretory phenotype (SASP), and age-related fibrosis in kidneys. Bmi1 deficiency impaired renal structure and function, increasing serum creatinine/urea, reducing creatinine clearance, and decreasing cortical thickness and glomerular number. However, knockout of the serine-threonine kinase Chk2 alleviated these aging phenotypes. Transcriptomics identified transforming growth factor beta 1 (TGFβ1) upregulation in Bmi1-deficient RTECs, but TGFβ1 was downregulated upon Chk2 knockout. The tumor suppressor protein p53 transcriptionally activated TGFβ1, promoting EMT in RTECs. Bmi1 knockout or oxidative stress (induced with H2O2) increased TGFβ1 expression, and EMT in RTECs and was partly reversed by p53 inhibition. Together, Bmi1 deficiency causes oxidative stress and DDR-mediated RTEC senescence/SASP, thus activating p53 and TGFβ1 to induce EMT and age-related fibrosis. However, blocking DDR (via Chk2 knockout) or p53 ameliorates these changes. Our study reveals mechanisms whereby Bmi1 preserves renal structure and function during aging by suppressing DDR and p53/TGFβ1-mediated EMT. These pathways represent potential targets for detecting and attenuating age-related renal decline.

Synergistic therapeutic approach for hemorrhoids: integrating mesenchymal stem cells with diosmin-hesperidin to target tissue edema and inflammation.

Mesenchymal stem cells (MSCs) have promising regenerative properties in tissue repair and anti-inflammatory responses. This study aimed to investigate the effects of MSCs and their combination with micronized purified flavonoid fraction (MPFF) in a croton oil-induced hemorrhoids model on tissue edema, inflammation, and underlying molecular mechanisms. MSCs were isolated and characterized for their adherence, differentiation capacity, and immunophenotyping. Croton oil-induced hemorrhoid mouse models were established to assess tissue edema, inflammation, tumor necrosis factor (TNF-α) expression, transforming growth factor-β (TGF-β) expression, collagen ratio, and MMP-9 activity. The effects of MSCs and their combination with MPFF (diosmin-hesperidin) were evaluated through histological examinations, western blot analysis, and gelatin zymography. Characterization confirmed the MSCs' plastic adherence, osteogenic differentiation potential, and immunophenotype (positive for CD90 and CD29, negative for CD45 and CD31). Treatment with MSCs alone or in combination with MPFF significantly reduced tissue edema, inflammation, TNF-α expression, and MMP-9 activity. Additionally, MSCs increased TGF-β expression, and collagen type I/III ratio, and accelerated wound healing by resolving inflammation. These findings suggest that MSCs play a crucial role in modulating TNF-α, TGF-β, collagen remodeling, and MMP-9 activity, highlighting their promising role in hemorrhoid treatment and wound healing processes. Further research is warranted to fully elucidate the intricate mechanisms and optimize MSC-based therapies for clinical applications in hemorrhoidal disease management.

The Effect of Topical Gel Secretome Hypoxic Mesenchymal Stem Cells (SH-MSCs) on IL-6 and TGF- β Gene Expression (Experimental Study in Male Wistar Alopecia-Like Rat Models Induced by Fluconazole)

Alopecia induced by the accumulation of reactive oxygen species (ROS) stimulates changes in the immune and neuroendocrine systems. Hair follicles have dermal papillary cells (DPCs) that bind to corticotropin-releasing factor (CRF) receptors. Research shows that transforming growth factor (TGF-β) is one of the expressions that can be an indication of the influence of CRF. The effect of CRF itself is to cause hair loss. The purpose of this study was to determine the Secretome of Hypoxia Mesenchymal Stem Cells (SH-MSCs) against IL-6 expression and TGF-β expression. This study used experimentally in vivo with the Post Test Only Control Group Design method. The subjects of this study were male rats of Wistar strain which were divided into 4 treatments consisting of 1 healthy group, 1 negative control group, 1 treatment group with SH-MSCs dose 10% in a dose of 200 mg/rat/day gel, and 1 more treatment group with SH-MSCs dose 20% in a dose of 200 mg/rat/day. On the 21st day, IL-6 expression and TGF-β expression were examined. Data were analyzed using the One-Way ANOVA Test and the Kruskall-Wallis Test to determine the effect of each group. The One-Way ANOVA test showed the results of IL-6 expression (p = 0.000) showed a significant difference (p &lt; 0.05) and the Kruskall-Wallis test of TGF-β expression (p = 0.000) with SH-MSCs dose 200 was a significant difference (p &lt; 0.05). The administration of 10% and 20% doses of SH-MSCs in 200 mg gel may decrease IL-6 expression. Dosed SH-MSCs 20%increased TGF-β expression in alopecia-like rats. The development of the use of SH-MSCs is expected to be an alternative to alopecia therapy that is more beneficial and safe for the body.

Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa

Previous studies showed that Trichinella spiralis galectin (Tsgal) facilitates larval invasion of intestinal epithelium cells (IECs). However, IEC proteins binding with Tsgal were not identified, and the mechanism by which Tsgal promotes larval invasion is not clear. Toll-like receptors (TLRs) are protein receptors responsible for recognition of pathogens. The aim of this study was to investigate whether recombinant Tsgal (rTsgal) binds to TLR-4, activates inflammatory pathway in gut epithelium and mediates T. spiralis invasion. Indirect immunofluorescence (IIF), GST pull-down and co-immunoprecipitation (Co-IP) assays confirmed specific binding between rTsgal and TLR-4 in Caco-2 cells. qPCR and Western blotting showed that binding of rTsgal with TLR-4 up-regulated the TLR-4 transcription and expression in Caco-2 cells, and activated p-NF-κB p65 and p-ERK1/2. Activation of inflammatory pathway TLR-4/MAPK-NF-κB by rTsgal up-regulated pro-inflammatory cytokines (IL-1β and IL-6) and down-regulated anti-inflammatory cytokine TGF-β in Caco-2 cells, and induced intestinal inflammation. TAK-242 (TLR-4 inhibitor) and PDTC (NF-κB inhibitor) significantly inhibited the activation of TLR-4 and MAPK-NF-κB pathway. Moreover, the two inhibitors also inhibited IL-1β and IL-6 expression, and increased TGF-β expression in Caco-2 cells. In T. spiralis infected mice, the two inhibitors also inhibited the activation of TLR-4/MAPK-NF-κB pathway, ameliorated intestinal inflammation, impeded larval invasion of gut mucosa and reduced intestinal adult burdens. The results showed that rTsgal binding to TLR-4 in gut epithelium activated MAPK-NF-κB signaling pathway, induced the expression of TLR-4 and pro-inflammatory cytokines, and mediated larval invasion. Tsgal might be regarded as a candidate molecular target of vaccine against T. spiralis enteral invasive stage.

The Wound-Healing Activity of PEDOT-PSS in Animals.

This study evaluated the wound-healing activity of a polymer, Poly(3,4-ethylenedioxythiophene):poly-(styrene sulfonate) (PEDOT: PSS), and determined its mechanism based on angiogenic activity in a full-thickness excision wound model in Spraque Dawley (SD) rats. Administering PEDOT: PSS (1.6) 1.5 ppm at a dose of 50 mg/kg/day significantly improved wound healing in the SD rats on the eleventh day after the incision was created. PEDOT: PSS-treated animals presented no anti-inflammatory skin effects; however, there was an increase in angiogenic behavior. VEGF was found to be significantly elevated in the PEDOT: PSS-treated groups seven days post-incision. However, only a higher concentration of PEDOT: PSS increased TGF-β1 expression within the same time frame. Our results showed that PEDOT: PSS enhances wound healing activity, mainly in terms of its angiogenic effects. In this paper, we describe the highly conductive macromolecular material PEDOT: PSS, which demonstrated accelerated wound-healing activity in the animal incision model. The results will further provide information regarding the application of PEDOT: PSS as a dressing for medical use.

The potency of propolis extract as anti-inflammatory agent for stimulation of reparative dentin formation

Background: As an effort to treat a deep caries lesion with a diagnosis of reversible pulpitis, one of the treatment options is pulp capping. For decades, calcium hydroxide has been considered the most acceptable pulp capping material and has been routinely used in dental practice, but it also has some weakness. To overcome that, alternative materials are needed that can provide a better effect. Recently, a lot of research has been done using natural ingredients, including Propolis. Propolis has been known to have strong antimicrobial and anti inflammatory properties. Therefore, it is necessary to conduct a deeper study regarding the potency of propolis extract as an anti inflammatory agent for the stimulation of reparative dentin formation so that it can be considered to be used as a pulp capping material. Objective: The aim of this literature review is to highlight the potency of propolis extract as anti inflammatory agent in the natural stimulation of reparative dentin formation. Methods: The search strategy consist of an electronic research using databases such as PubMed, Google Scholar, Science Direct, dan Springer Link for studies published from 2003 to 2021 regarding the potency of propolis extract as anti inflammatory agent for stimulation of reparative dentin formation, including review articles and research articles. Results: Propolis and its constituents have been proven to be used as a potent natural anti inflammatory, it also considered to have the ability to promote reparative dentin formation. However, there are some literatures stated that reparative dentin does not form with the application of propolis, and the causes may vary. Conclusion: Propolis extract has a potency as an anti inflammatory agent in dental pulp inflammation by inhibiting TNFα expression, and increasing TGF-β1 expression, as well as its potency in stimulating the formation of reparative dentin. Based on this review, further studies are needed.

Carbon dioxide-induced decrease in extracellular pH enhances the production of extracellular matrix components by upregulating TGF-β1 expression via CREB activation in human dermal fibroblasts.

Mild acidification caused by transcutaneous administration of carbon dioxide (CO2 ) has been reported to improve some epidermal skin impairments, such as desquamation and inflammation; however, its effects on dermal tissue remain unclear. Here, we examined the effect and mechanism of mild acidity on extracellular matrix (ECM) protein production in normal human dermal fibroblasts (NHDFs). To achieve this, the skin permeability of CO2 and its effect on intradermal pH were evaluated by treating reconstructed human skin equivalents (HSEs) with a CO2 -containing formulation. Additionally, NHDFs were cultured in a pH-adjusted medium (pH 6.5). CO2 successfully permeated HSEs and reduced the intradermal pH. Decreased extracellular pH activated CREB, upregulated TGF-β1 expression, promoted the production of elastic and collagen fibres, and increased hyaluronan concentration in NHDFs. Additionally, the low pH-induced increase in TGF-β1 expression was attenuated via the RNAi-mediated suppression of the expression of CREB1 and proton-sensing G protein-coupled receptors (GPCRs), including GPR4 and GPR65. Moreover, low pH-induced CREB activation was suppressed by the inhibition of the cAMP/PKA and PLC/PKC signalling pathways. Taken together, a CO2 -induced decrease in intradermal pH may promote ECM production in NHDFs via the upregulation of TGF-β1 expression, which was mediated by the activation of the GPCR signalling pathway and CREB, indicating that CO2 could be used to treat ultraviolet radiation-induced photoaging, intrinsic ageing and ECM deterioration.

Peppermint Leaf Extract Cream Increased Transforming Growth Factor β (TGF-β) Expression and Collagen Amount In The Male Wistar Rat's Skin Exposed to UVB

Peppermint leaves contain flavonoids, menthofuran, and other phenolic compounds that have antioxidant activity and are believed to inhibit collagen degradation in the skin. This study investigated the efficacy of a 5% peppermint leaf extract cream in increasing the transforming growth factor β (TGF-β) expression and collagen amount as indicators of collagen maintenance in male Wistar rats' skin exposed to UVB. Method A randomized post-test-only control group design was performed on male Wistar rats, local strains, aged 2-3 months, with body weights ranging from 180 to 200 grams. According to the sample calculation, 21 rats were randomly divided into three groups. The first group is the control group (without any treatment). The second group was treated with UVB without cream (UVB-only group), and the third group was treated with UVB + 5% peppermint leaf extract cream on exposed skin. After four weeks of treatment, the tissue samples were examined. The expression of TGF-β was evaluated by immunohistochemical examination, and the amount of collagen was examined by Picro-Sirius red staining. Comparative tests were performed on the measurement results of the three groups. The results showed a significant difference in the mean TGF-β expression and collagen amount among the three groups (p&lt;0.001). The group exposed to UVB + 5% peppermint leaf extract cream had the highest mean TGF-β expression and collagen amount (77.41±5.79% and 82.57±2.18%, respectively), followed by the control group (63.17±12.22% and 74.05±7.15%) and the UVB-only group (50.54±11.10% and 65.65±5.56%). Administration of 5% peppermint leaf extract cream effectively increased TGF-β expression and collagen amount in the skin of male Wistar rats exposed to UVB compared to the control and the UVB-only groups.

Abstract 4850: High salt diet induced tumor initiating stem cells mediate breast cancer progression

Abstract High-salt (sodium chloride) diets have been associated with several chronic inflammatory diseases. While the role of chronic inflammation in cancer is well established, the specific role of high salt diet in carcinogenesis is unknown. Previous studies with syngeneic murine breast cancer models, both in our laboratory and others, have shown that high salt (HS) diet induced tumor regression through inflammatory activation of anti-tumor adaptive immune responses. We tested this counter-intuitive and suspiciously beneficial role of HS diet by performing sequential passage studies in preclinical murine models. Six week old mice were placed on high salt diet for 2 weeks prior to injection with syngeneic 4T1 and Py230 triple negative breast cancer cells (into BALB/c and C57Bl/6J mice, respectively, referred to as passage-1). Tumors were harvested after four weeks of injection. The cancer cells depleted of immune cells were collected from these harvested tumors and reinjected into new non-tumor bearing mice. This cycle was repeated three times. In passage-1 cohort, by day 28, HS diet induced reduced tumor progression in both 4T1-BALB/c (267 ± 59 mm3) and Py230-C57Bl/6J (238 ± 54 mm3) as compared to regular salt (RS) diet cohort (611 ± 94 mm3 and 473 ± 69 mm3, respectively; p&amp;lt;0.05 for both). However, in passage-4 cohort, by day 28, HS diet induced significantly higher tumor progression in both 4T1-BALB/c (806 ± 91 mm3) and Py230-C57Bl/6J (743 ± 81 mm3) models, as compared to regular salt (RS) diet cohort (577 ± 83 mm3 and 462 ± 77 mm3, respectively; p&amp;lt;0.05 for both). Cellular analysis by flow cytometry revealed that there was a 12-19 fold increase in tumor initiating stems cells (TISCs) in passage-4 HS diet cohort compared to RS diet cohort. Mechanistic studies have demonstrated that there was increased TGFβ expression on TISCs obtained from passage-4 HS diet cohort which correlated with enhanced exhaustion phenotype (CTLA4+) of tumor infiltrating adaptive immune cells (CD4 and CD8 T cells) in this cohort. Co-treatment with anti-TGFβ and anti-CTLA4 monoclonal antibodies (mAb) in passage-4 HS diet cohort, significantly reduced tumor progression (p&amp;lt;0.05), as compared to treatment with either mAb alone. Taken together, these data demonstrated that chronic HS diet leads to expansion of TGFβ expressing TISCs leading to host immune exhaustion and tumorigenesis. Importantly, anti-TGFβ mAb exerted a favorable synergistic effect to enhance the anti-tumor efficacy of immune check-point inhibitors. Citation Format: Lisa Tucker, Sonya Reid, Jeffrey C. Rathmell, Venkataswarup Tiriveedhi. High salt diet induced tumor initiating stem cells mediate breast cancer progression. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4850.