Activation Of TGF-β1 Signaling Pathway Research Articles

Role of AP-2α/TGF-β1/Smad3 axis in rats with intervertebral disc degeneration

ObjectiveStudies have proposed the role of AP-2α in human disease. However, few have focused on its effects on intervertebral disc degeneration (IDD). This study intends to discuss the role of AP-2α in IDD by regulating TGF-β1 and Smad3 expression. MethodsThe AP-2α and TGF-β1 expression in IDD NP clinical samples was detected. Rat models of IDD were established by acupuncture. The rats were injected with AP-2α low expression adeno-associated virus or TGF-β1 high expression adeno-associated virus to observe their effects on pathological damages, NP cell apoptosis, matrix metalloproteinase-2 (MMP-2), MMP-9, Smad3, Aggrecan and collagen (Col)-2 expression in NP tissues. The NP cells were isolated and transfected with silenced AP-2α or overexpressed TGF-β1 vector to figure out their functions in growth, senescence and apoptosis. ResultsAP-2α and TGF-β1 were upregulated in NP tissues of patients and rats with IDD. AP-2α silencing limited the activation of TGF-β1 signaling pathway. Reduced AP-2α ameliorated pathological changes, declined MMP-2, MMP-9 and Smad3 expression and elevated Aggrecan and Col-2 expression in NP tissues of rats with IDD, and speeded up the growth and depressed senescence and apoptosis of NP cells of rats with IDD. Up-regulating TGF-β1 weakened the effect of down-regulated AP-2α on NP tissues and cells in IDD. ConclusionCollectively, our study demonstrates that knockdown of AP-2α restricts TGF-β1 and Smad3 expression to promote proliferation and depress senescence and apoptosis of NP cells in rats with IDD.

Abstract 4395: Activation of TGFβ1 signaling pathway in ovarian cancer stem cells plays critical role in ovarian cancer tumorigenesis

Abstract Ovarian cancer (OC) is the most lethal gynecological malignancy worldwide, primarily due to lack of a reliable early diagnostic method and high incidence of chemo-resistance induced tumor recurrence and relapse. Development of early screening and diagnosis biomarkers is the key for survival improvement of patients with OC. In this study, we applied a system biology approach to identify dysregulated genes and pathways during OC tumorigenesis, as well as to study the roles of OC stem cells (OCSCs) during this process. In order to identify diagnostic/therapeutic biomarkers for OC, gene expression between OC patient malignant tumors and normal tissues were compared with datasets GSE4122 and GSE26712. 188 and 305 genes were found to be consistently downregulated and upregulated respectively (fold change ≥2, and FDR<0.05) across the two datasets. Among the dysregulated genes, OCSC biomarkers CD24 and PROM1, and cancer cell stemness related genes such as SOX9, EPCAM, HMGA1, EZH2, BIRC5, and KRT7 were all significantly upregulated in OC. In contrast, many metabolism related genes were downregulated in OC, such as ABCA8, ACOX2, PFKP, ADH1B, and PKD2. Among the upregulated genes, over 30 genes were found to be significantly associated with tumor stemness scores (r>0.40 and P<.0001). The top associated genes such as CCNB2, NDC80, TTK, CDK1, NEK2, RAD54L, CDC20, KIF23, BIRC5, EZH2 (r>0.40) were all previously reported to be associated with aggressive disease and poor diagnosis in different cancer types. OCSCs were considerd to play critical roles in OC tumour initiation, dissemination, metastasis and recurrence. To elucidate the role of OCSCs during tumorigenesis, gene expression were compared between OCSCs and normal stem cells using dataset GSE90125. Interestingly, tumor microenvironment and EMT related genes such as IL33, WISP1, PRG2, FAP, ENPP2, and SBSN were found to be among the most upregulated genes in OCSCs, indicating important role of tumor microenvironment during the conversion of normal stem cells to tumor stem cells. Moreover, pathway analysis with the dysregulated genes showed that TGFβ1 signaling pathway was significantly activated in both OC tumors and OCSCs. Previous study showed that TGFβ1 signaling pathway promotes OC tumor growth by upregulating VCAN (upregulated in OC) in the tumor microenvironment. Therefore, we conclude that the modulation of tumor microenvironment through the activated TGFβ1 signaling pathway in OCSCs plays important roles during OC tumorigenesis and may be used as diagnostic and/or therapeutic targets for patients with OC. Citation Format: Xiaoli Zhang, Shuai Shao, Lang Li, Qi-En Wang. Activation of TGFβ1 signaling pathway in ovarian cancer stem cells plays critical role in ovarian cancer tumorigenesis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4395.

Potential ameliorative effects of epigallocatechin-3-gallate against cigarette smoke exposure induced renal and hepatic deficits.

The environmental pollution caused by cigarette smoke (CS) seriously endangers people's health. Epigallocatechin-3-gallate (EGCG) is the most abundant catechin in green tea. In this study, rats were exposed to CS for 90 days. Kidney function was evaluated by detecting the levels of serum creatinine and blood urea nitrogen. Liver function was evaluated by detecting the activities of alanine aminotransferase and aspartate transaminase. The renal and hepatic oxidative stress and inflammation were assessed by detecting the levels of malondialdehyde, reduced glutathione, antioxidant enzymes (superoxide dismutase and glutathione peroxidase) and proinflammatory cytokines. Organ fibrosis was evaluated by observing collagen deposition via masson staining, by examining the hydroxyproline level, by measuring the mRNA levels of fibrosis-associated genes collagen (Col)-1A1 and Col-3A1, as well as by assessing the activity of profibrotic TGF-β1 pathway. Additionally, renal and hepatic epithelial-mesenchymal transition (EMT) were evaluated. It was observed that EGCG ameliorated the renal and hepatic oxidative stress, inflammation, EMT, as well as inhibited the activation of TGF-β1 signaling pathway induced by CS. These results showed that EGCG could attenuate CS-induced renal and hepatic fibrosis.

Huangqi Shengmai Yin Protects against Radiation-Induced Cardiac Fibrosis Injury by Regulating the TGF-β1/Smads and MMPs.

Background Radiation-induced heart damage is considered to be a progressive process of fibrosis. Emerging evidence has indicated that the Smads and matrix metalloproteinases (MMPs)/tissue inhibitors of MMPs (TIMP) may be involved in radiation-induced cardiac fibrosis (RICF) by regulating the activation of TGF-β1 signaling pathway. Based on this, the present study was undertaken to characterize the effect of Huangqi Shengmai Yin (HSY) on RICF in a rat model. Methods Precardiac region of rats was irradiated with 25 Gy X-rays, and their myocardial pathology scores in terms of injury and collagen volume fraction (CVF) and the expression levels of fibrotic molecules were detected. Results The pathology scores and CVF in myocardial tissue increased after irradiation, and the expression of TGF-β1, Col1, and Col3 increased. This change indicated that such irradiation promoted the fibrosis damage in rat hearts. The damage was accompanied by an increase in the expression of Smad 2, Smad3, Smad4, and MMP14 and a decrease in the expression of Smad 7 and TIMP1. Administration of HSY weakened the RICF by decreasing pathology score and CVF and decreased the expression of TGF-β1, Col1, and Col3 in irradiated rat hearts. In addition, Smad2, Smad3, Smad4, and MMP14 were downregulated, while Smad 7 and TIMP1 were upregulated during intervention with HSY. Conclusions The involvement of the TGF-β1/Smads and MMPs/TIMP system in RICF was confirmed. This study demonstrated, for the first time, that HSY attenuates the effects of RICF in a rat model. The effect HSY was found to be closely related to the TGF-β1/Smads signaling pathway and MMPs system. These results suggest that HSY is a prospective drug for clinical treatment of RICF.

Regulation of IL-17A and implications for TGF-β1 comodulation of airway smooth muscle remodeling in severe asthma.

Severe asthma develops as a result of heightened, persistent symptoms that generally coincide with pronounced neutrophilic airway inflammation. In individuals with severe asthma, symptoms are poorly controlled by high-dose inhaled glucocorticoids and often lead to elevated morbidity and mortality rates that underscore the necessity for novel drug target identification that overcomes limitations in disease management. Many incidences of severe asthma are mechanistically associated with T helper 17 (TH17) cell-derived cytokines and immune factors that mediate neutrophilic influx to the airways. TH17-secreted interleukin-17A (IL-17A) is an independent risk factor for severe asthma that impacts airway smooth muscle (ASM) remodeling. TH17-derived cytokines and diverse immune mediators further interact with structural cells of the airway to induce pathophysiological processes that impact ASM functionality. Transforming growth factor-β1 (TGF-β1) is a pivotal mediator involved in airway remodeling that correlates with enhanced TH17 activity in individuals with severe asthma and is essential to TH17 differentiation and IL-17A production. IL-17A can also reciprocally enhance activation of TGF-β1 signaling pathways, whereas combined TH1/TH17 or TH2/TH17 immune responses may additively impact asthma severity. This review seeks to provide a comprehensive summary of cytokine-driven T cell fate determination and TH17-mediated airway inflammation. It will further review the evidence demonstrating the extent to which IL-17A interacts with various immune factors, specifically TGF-β1, to contribute to ASM remodeling and altered function in TH17-driven endotypes of severe asthma.

Upregulation of lincRNA-p21 in thoracic aortic aneurysms is involved in the regulation of proliferation and apoptosis of vascular smooth muscle cells by activating TGF-β1 signaling pathway.

Long intergenic noncoding RNA-p21 (lincRNA-p21) has been proved in the pathogenesis of aortic aneurysms, while its functionality in thoracic aortic aneurysms (TAA) and the mechanism of function remains unclear. Therefore our study aimed to investigate the role of lincRNA-p21 in TAA. Aortic media specimens and blood samples were collected from both patients with TAA and healthy controls. Expression of lincRNA-p21 in those tissues was detected by reverse-transcription quantitative polymerase chain reaction (qRT-PCR). Diagnostic values of lincRNA-p21 in aortic media and blood for TAA were evaluated by receiver operating characteristic curve analysis. LincRNA-p21 overexpression human vascular smooth muscle cells (VSMCs) were prepared and the effects of lincRNA-p21 overexpression on cell proliferation and apoptosis were explored by cell counting kit-8 assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, respectively. Expression of lincRNA-p21 and transforming growth factor β1 (TGF-β1) in VSMCs with different treatment was detected by qRT-PCR and Western blot analysis, respectively. Expression of lincRNA-p21 in aortic media tissues and blood was significantly upregulated in TAA patients than in healthy controls. Expression of lincRNA-p21 in aortic media and blood can be used to effectively distinguish TAA patients form healthy controls. LincRNA-p21 overexpression inhibited proliferation and promoted apoptosis of VSMCs, while TGF-β1 inhibitor reduced those effects. LincRNA-p21 overexpression upregulated TGF-β1 expression, while TGF-β1 activator showed no significant effects on lincRNA-p21 expression in VSMC. LincRNA-p21 participates in TAA by regulating the proliferation and apoptosis of VSMCs through the activation of TGF-β1 signaling pathway.

Sodium Tanshinone IIA Sulfonate Prevents Angiotensin II-Induced Differentiation of Human Atrial Fibroblasts into Myofibroblasts.

Differentiation of atrial fibroblasts into myofibroblasts plays a critical role in atrial fibrosis. Sodium tanshinone IIA sulfonate (DS-201), a water-soluble derivative of tanshinone IIA, has been shown to have potent antifibrotic properties. However, the protective effects of DS-201 on angiotensin II- (Ang II-) induced differentiation of atrial fibroblasts into myofibroblasts remain to be elucidated. In this study, human atrial fibroblasts were stimulated with Ang II in the presence or absence of DS-201. Then, α-smooth muscle actin (α-SMA), collagen I, and collagen III expression and reactive oxygen species (ROS) generation were measured. The expression of transforming growth factor-β1 (TGF-β1) and the downstream signaling of TGF-β1, such as phosphorylation of Smad2/3, were also determined. The results demonstrated that DS-201 significantly prevented Ang II-induced human atrial fibroblast migration and decreased Ang II-induced α-SMA, collagen I, and collagen III expression. Furthermore, increased production of ROS and expression of TGF-β1 stimulated by Ang II were also significantly inhibited by DS-201. Consistent with these results, DS-201 significantly inhibited Ang II-evoked Smad2/3 phosphorylation and periostin expression. These results and the experiments involving N-acetyl cysteine (antioxidant) and an anti-TGF-β1 antibody suggest that DS-201 prevent Ang II-induced differentiation of atrial fibroblasts to myofibroblasts, at least in part, through suppressing oxidative stress and inhibiting the activation of TGF-β1 signaling pathway. All of these data indicate the potential utility of DS-201 for the treatment of cardiac fibrosis.

The Effects of Different Fluences of 1064 nm Q-Switched Nd:YAG Laser on Skin Repair and Skin Barrier Dysfunction in Mice.

The purpose of this study was to investigate the effects of different fluences of Q-switched 1064 nm Nd:YAG laser on skin repair and barrier, and clarify its mechanisms. The Q-switched 1064 nm Nd:YAG laser is widely used for rejuvenation, which needs appropriate fluence data to optimize efficacy and minimize side effects, and for elucidation of action mechanism. The dorsal skin of BABL/c mice was administered 0, 1, 1.5, and 2 J/cm2 energy level laser, twice a week for 4 weeks. Immediately, 7, 14, 21, and 28 days after last treatment, the skin elasticity, moisture content, and transepidermal water loss (TEWL) were measured; 7, 14, 21, and 28 days after last treatment, the hydroxyproline content, mRNA level of procollagen types I and III, protein level of keratin-10 (K-10), filaggrin, transforming growth factor beta receptor II (TGFβRII), Smad2, and p65 were detected. Compared with the unirradiated control, the laser treatments decreased skin elasticity immediately, but increased skin moisture content in the 2 J/cm2 group, and then from day 21 to day 28, the skin elasticity, moisture content, hydroxyproline content, and gene expression of types I and III procollagen increased significantly. The TEWL value of the irradiated group significantly increased after irradiation immediately and 7 days after, K-10 and filaggrin were also decreased at 7 days after. The phosphorylation of TGFβRII (p-TGFβRII) increased at days 7 and 21, and phosphorylation of Smad2 (p-Smad2) was induced at 21 days. Irradiation of 1064 nm Q-switched Nd:YAG laser was able to markedly promote repair of mouse skin within 28 days through stimulation of collagen synthesis, with less skin barrier dysfunction, especially at the 1.5 J/cm2 fluence, and the activation of TGFβ1-signaling pathways seemed to play an important role in repair.

Activation of TGFβ-1 signaling pathways in human breast cancer cell line MDA-MB-231 induced by hexachlorobenzene pesticide

Activation of TGFβ-1 signaling pathways in human breast cancer cell line MDA-MB-231 induced by hexachlorobenzene pesticide

Role of serotonergic system in the pathogenesis of fibrosis in canine idiopathic inflammatory myopathies

Idiopathic inflammatory myopathies are muscle diseases characterized by inflammation, necrosis, and fibrosis. The neurotransmitter serotonin (5-HT) has been shown to promote fibrosis in many tissues and organs by activating TGFβ-1 signaling. In this study, we evaluated the potential role of 5-HT in the pathogenesis of fibrosis in canine idiopathic inflammatory myopathies. Muscle biopsies from dogs affected by masticatory muscle myositis or polymyositis and from healthy dogs were processed for immunohistochemistry and Western blotting. The immunohistochemical analysis showed a strong expression of 5-HT in muscle tissues of affected dogs, whereas the amine was absent in the muscles of healthy dogs. Biochemical analysis showed increased expression levels of the selective 5-HT2A receptor in the muscle specimens of the most severely affected dogs versus controls. Further, increased phosphorylation levels of the TGFβ-1 signaling mediators SMAD2/3 and ERK1/2 were detected in tissue samples from affected dogs as compared to tissues from healthy dogs. Although further studies are needed, our findings highlight for the first time a potential role of 5-HT in the development of fibrosis in canine idiopathic inflammatory myopathies, thus supporting other evidence that 5-HT pro-fibrotic activity occurs via activation of TGFβ-1 signaling pathway.

Sevoflurane-mediated TGF-β1 signaling in renal proximal tubule cells

Several volatile anesthetics, including sevoflurane, protect against renal ischemia-reperfusion injury in vivo by reducing necrosis and inflammation. Furthermore, in cultured renal tubule cells, sevoflurane directly induced the phosphorylation of the cytoprotective kinases (ERK and Akt), upregulated 70-kDa heat shock protein (HSP70), and attenuated nuclear translocation of the proinflammatory transcription factor NF-kappaB. It has been shown that sevoflurane increases the release of transforming growth factor-beta1 (TGF-beta1) in human proximal tubule (HK-2) cells via externalization of plasma membrane phosphatidylserine (PS), and this increase in TGF-beta1 protected HK-2 cells against hydrogen peroxide-mediated necrosis. In this study, we aimed to determine whether the sevoflurane-mediated phosphorylation of ERK and Akt, induction of HSP70, and reduction in NF-kappaB activation are due to TGF-beta1 receptor-mediated signaling after PS externalization in HK-2 cells. Exogenous TGF-beta1 and a liposome mixture containing PS mimicked sevoflurane-mediated ERK and Akt phosphorylation and HSP70 induction in HK-2 cells. Sevoflurane and TGF-beta1 caused the nuclear translocation of the SMAD3 transcription factor in HK-2 cells. Furthermore, a neutralizing TGF-beta1 antibody or exogenous annexin V to bind PS prevented sevoflurane-induced ERK and Akt phosphorylation and HSP70 induction in HK-2 cells. Finally, a TGF-beta1 antibody and annexin V attenuated the reduction in nuclear translocation of NF-kappaB by sevoflurane. Therefore, we demonstrate in this study that sevoflurane-mediated cytoprotective and anti-inflammatory effects in HK-2 cells are at least partially due to the externalization of PS and activation of TGF-beta1 signaling pathways.