Bone morphogenetic proteins (BMPs) are the largest subfamily of the transforming growth factor-β superfamily, and they play important roles in the development of numerous organs, including the inner ear. The inner ear is a relatively small organ but has a highly complex structure and is involved in both hearing and balance. Here, we discuss BMPs and BMP signaling pathways and then focus on the role of BMP signal pathway regulation in the development of the inner ear and the implications this has for the treatment of human hearing loss and balance dysfunction.
Body Patterning ; Bone Morphogenetic Protein Receptors/physiology* ; Bone Morphogenetic Proteins/physiology* ; Cell Differentiation ; Cochlea/embryology* ; Ear, Inner/embryology* ; Hedgehog Proteins/physiology* ; Humans ; Signal Transduction/physiology* ; Smad Proteins/physiology* ; Vestibule, Labyrinth/embryology* ; Wnt Signaling Pathway

OBJECTIVETo investigate the effects of YOD1 overexpression on the proliferation and migration of human oral keratinocytes (HOKs), and to clarify whether the mechanisms involve transforming growth factor-β (TGF-β) signaling.

METHODSHOKs were transfected with the plasmid pEGFP-N3-YOD1 containing YOD1. The mRNA levels of YOD1 and TGF-β were determined by qPCR. The protein expressions of YOD1, TGF-β, Smad2/3, Smad4, and phospho-Smad2/3 were determined by western blotting. Cell proliferation and migration were evaluated by Cell Counting Kit-8 assay and wound healing assay, respectively.

RESULTSThe mRNA and protein levels of YOD1 were higher in HOKs transfected with YOD1. YOD1 overexpression significantly enhanced the migration of HOKs. The mRNA and protein levels of TGF-β3 were increased by YOD1 overexpression. HOKs transfected with YOD1 exhibited increased phospho-Smad2/3 levels.

CONCLUSIONYOD1 overexpression enhances cell migration by promoting TGF-β3 signaling which may play an important role in lip and palate formation. YOD1 mutation may contribute to aberrant TGF-β3 signaling associated with decreased cell migration resulting in NSCLP.

Cell Movement ; physiology ; Cell Proliferation ; Cells, Cultured ; Endopeptidases ; genetics ; metabolism ; Humans ; Keratinocytes ; physiology ; Signal Transduction ; physiology ; Smad Proteins ; genetics ; metabolism ; Thiolester Hydrolases ; genetics ; metabolism ; Transforming Growth Factor beta3 ; genetics ; metabolism

Liver fibrosis is a common pathological process for chronic liver injury caused by multiple etiological factors and an inevitable phase leading to liver cirrhosis. According to the previous studies, hesperidin (HDN) shows a very good protective effect on CCl4-induced chemical hepatic fibrosis in rats. In this experiment, based on the findings of the previous studies, a platelet-derived growth factor (PDGF)-induced HSC-T6 model was established to observe the inhibitory effect of HDN on HSC-T6 proliferation. The ELISA method was adopted to detect the content of collagen I in HSC-T6 supernatant. Transforming growth factor (TGF)-beta1, Smad2, Smad3, Smad7 and connective tissue growth factor (CTGF) mRNA expressions were measured by RT-PCR; TGF-beta1 and CT-GF protein expressions in HSC-T6 were determined by Western blot, in order to study HDN's effect on TGF-beta1 signaling pathway in HSC and its potential action mechanism. The results demonstrated that HDN could notably improve HSC-T6 proliferation, Collagen I growth and TGF-beta1, Smad2, Smad3 and CTGF mRNA.expressions. After being intervened with HDN, it could notably inhibit HSC-T6 proliferation and Collagen I growth, reduce TGF-beta1, Smad2, Smad3 and CTGF mRNA and TGF-beta1, CTGF protein expressions and increase Smad7 mRNA expression. HDN's antihepatic fibrosis effect may be related to the inhibition of HSC proliferation and activation by modulating TGF-beta/Smad signaling pathway.
Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Connective Tissue Growth Factor ; physiology ; Hesperidin ; pharmacology ; Platelet-Derived Growth Factor ; pharmacology ; Rats ; Signal Transduction ; drug effects ; Smad Proteins ; physiology ; Transforming Growth Factor beta1 ; physiology

OBJECTIVETo investigate the effects and mechanisms of cordycepin,an effective component of cordyceps militaris, on renal interstitial fibrosis (RIF) and its related eIF2α/TGF-β/Smad signaling pathway.

METHODFirstly, 15 C57BL/6 mice were randomly divided into 3 groups,the control group (Group A), the model group (Group B) and the cordycepin-treated group (Group C). After renal interstitial fibrotic model was successfully established by unilateral ureteral obstruction (UUO), the mice in Group C were intraperitoneally administrated with cordycepin(5 mg x kg(-1) d(-1)) and the ones in Group A and B were administrated with physiological saline for 5 days. At the end of the study, the obstructed kidneys were collected and detected for the pathological changes of RIF, and the mRNA expressions of collagen type I (Col I) and α-smooth muscle actin (α-SMA) in the kidney by Northern blot. Secondly, after renal tubular epithelial (NRK-52E) cells cultured in vitro were exposed to transforming growth factor (TGF) -β with or without cordycepin, the mRNA expressions of Col I and collagen type IV( Col IV) by Northern blot, and the protein expressions of eukaryotic initiation factor 2α (eIF2α), phosphorylated eIF2α ( p-eIF2α), Smad2/3 and phosphorylated Smad2/3 (p-Smad2/3) were tested by Western blot.

RESULTIn vivo, cordycepin alleviated RIF in model mice, including improving fibrotic pathological characteristics and mRNA expressions of Col I and α-SMA. In vitro, cordycepin induced the high expression of p-elF2α, and inhibited the expressions of p-Smad2/3, Col I and Col IV induced by TGF-β in NRK-52E cells.

CONCLUSIONCordycepin attenuates RIF in vivo and in vitro, probably by inducing the phosphorylation of eIF2α, suppressing the expression of p-Smad2/3, a key signaling molecule in TGF-β/Smad signaling pathway, and reducing the expressions of collagens and α-SMA in the kidney.

Actins ; analysis ; Animals ; Deoxyadenosines ; pharmacology ; Fibrosis ; Kidney ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Phosphorylation ; Protein-Serine-Threonine Kinases ; physiology ; Signal Transduction ; drug effects ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; antagonists & inhibitors ; physiology

Transforming growth factor-β (TGF-β) members are key cytokines that control embryogenesis and tissue homeostasis via transmembrane TGF-β type II (TβR II) and type I (TβRI) and serine/threonine kinases receptors. Aberrant activation of TGF-β signaling leads to diseases, including cancer. In advanced cancer, the TGF-β/SMAD pathway can act as an oncogenic factor driving tumor cell invasion and metastasis, and thus is considered to be a therapeutic target. The activity of TGF-β/SMAD pathway is known to be regulated by ubiquitination at multiple levels. As ubiquitination is reversible, emerging studies have uncovered key roles for ubiquitin-removals on TGF-β signaling components by deubiquitinating enzymes (DUBs). In this paper, we summarize the latest findings on the DUBs that control the activity of the TGF-β signaling pathway. The regulatory roles of these DUBs as a driving force for cancer progression as well as their underlying working mechanisms are also discussed.
Animals ; Humans ; Molecular Targeted Therapy ; Receptors, Transforming Growth Factor beta ; metabolism ; Signal Transduction ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; physiology ; Ubiquitin Thiolesterase ; metabolism ; Ubiquitin-Specific Proteases ; Ubiquitination

BACKGROUNDAlthough various systemic and local factors such as abnormal carbohydrate or calcium metabolism, aging, and hormonal disturbances have been suggested as causes of ossification of the posterior longitudinal ligament (OPLL), the etiology of OPLL is not fully understood. The purpose of this study was to investigate whether bone morphogenetic protein (BMP)-2 is a candidate gene to modify the susceptibility of OPLL and the mechanism of signal transduction in ossification.

METHODSA total of 420 OPLL patients and 506 age- and sex-matched controls were studied. The complete coding sequence of the human BMP-2 gene was analyzed using polymerase chain reaction (PCR) and direct sequencing. All single nucleotide polymorphisms (SNPs) were detected and genotyped. BMP-2 expression vectors containing positive polymorphisms were constructed and transfected into the C3H10T1/2 cells. The expression of BMP-2 and the Smad signal pathway in positive cell clones were detected by Western blotting. The alkaline phosphatase (ALP) activity was determined using quantitative detection kits.

RESULTSThe frequencies for the 109T > G and 570A > T polymorphisms were different between the case and control groups. The "TG" genotype in 109T > G polymorphism is associated with the occurrence of OPLL, the frequency of the "G" allele is significantly higher in patients with OPLL than in control subjects (P < 0.001). The "AT" genotype in 570A > T polymorphism is associated with the occurrence of OPLL, the frequency of the "T" allele is significantly higher in patients with OPLL than in control subjects (P = 0.005). Western blotting analysis revealed that the expression of P-Smad1/5/8 protein transfected by wild-type or mutant expression vectors were significantly higher than control groups (P < 0.05), but there was no statistical difference in each experimental group (P > 0.05). The expression of Smad4 protein transfected by wild-type or mutant expression vectors was significantly higher than control groups (P < 0.05). The expression of Smad4 protein transfected by pcDNA3.1-BMP2 (109G) and pcDNA3.1-BMP2 (109G, 570T) was significantly higher than the other experimental groups (P < 0.05). The increase in ALP activity has been detected in pcDNA3.1-BMP2 (109G) and pcDNA3.1-BMP2 (109G, 570T) transfected cells up to 4 weeks after stable transfection. Activity of ALP was (30.56 ± 0.46) nmol×min(-1)×mg(-1) protein and (29.62 ± 0.68) nmol×min(-1)×mg(-1) protein, respectively. This was statistically different compared with the other experimental groups (P < 0.05).

CONCLUSIONSBMP-2 is the predisposing gene of OPLL. The "TG" genotype in the 109T > G and the "AT" genotype in the 570A > T polymorphisms are associated with the occurrence of OPLL. The 109T > G polymorphism in exon-2 of the BMP-2 gene is positively associated with the level of Smad4 protein expression and the activity of ALP. The Smad mediated signaling pathway plays an important role during the pathological process of OPLL induced by SNPs of BMP-2 gene.

Adult ; Aged ; Bone Morphogenetic Protein 2 ; genetics ; Cells, Cultured ; Female ; Humans ; In Situ Hybridization ; Longitudinal Ligaments ; metabolism ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Signal Transduction ; genetics ; physiology ; Smad Proteins ; metabolism ; Spine ; metabolism

OBJECTIVETo study the protective effect of oxymatrine (OMT) on myocardial fibrosis induced by acute myocardial infarction in rats and its effect on TGF-beta-Smads signal pathway.

METHODArteria coronaria ligation-induced acute myocardial infarction model was established in rats. The survived rats were randomly allotted into the model group, 50, 25, 12.5 mg x kg(-1) OMT groups, the 50 mg x kg(-1) captopril group, and the Sham-operated group which was treated as the model group without the arteria coranaria ligation. After 8 weeks of ligation, myocardial fibrosis was detected by HE and Masson staining, and the RT-PCR method were used to detect the expression of mRNA of TGF-beta-Smads signal system.

RESULTThe histopathological examination showed decrease in cardiocytes, deposition of extra-cellular matrix, and increase of collagen contents after 8 weeks of ligation. RT-PCR results showed that mRNA expressions of TGF-beta1, TbetaR1, Smad2, Smad3 and Smad4 significantly increased, but mRNA expression of Smad7 is remarkable lower than the sham-operated group. Treatment with OMT for 8 weeks could remarkably inhibit myocardial fibrosis, decrease mRNA expressions of TGF-beta1, TbetaR1, Smad2, Smad3, and Smad4, and increase mRNA expressions of Smad7.

CONCLUSIONOMT has the inhibitory effect on the experimental myocardial fibrosis induced by AMI in rats. Its mechanism may be closely related to TGF-beta-Smads signal system.

Acute Disease ; Alkaloids ; therapeutic use ; Animals ; Fibrosis ; Male ; Myocardial Infarction ; complications ; Myocardium ; pathology ; Quinolizines ; therapeutic use ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; physiology ; Smad Proteins ; genetics ; physiology ; Transforming Growth Factor beta ; genetics ; physiology

Bone morphogenetic proteins (BMPs) belong to TGF-β superfamily and are a group of important cytokines involved in cell differentiation, proliferation and embryonic development. Multiple BMPs play important roles in several functions of vertebrates. Signaling pathway of BMPs is known to be mediated by Smad proteins, which include 8 members while Smad1, Smad5 and Smad8 are involved in BMPs signal transduction while Smad2 and Smad3 are mediated TGF-β signal transduction. Although several BMPs such as BMP4 and BMP9 have been documented in the liver, BMP13 has not been examined in the liver. BMP13 also known as growth differentiation factor (GDF)-6 or cartilage-derived morphogenetic protein (CDMP)-2 is one of the BMPs family members. Function of BMP13 has been investigated in bone and tendon repair. It can stimulate tendon-like cell proliferation. However, our recent findings revealed that there was expression of BMP13 in the liver and its expression was modulated during metabolic disorders. The current article is to understand biological function of BMP13 especially in the liver.
Bone Morphogenetic Proteins ; metabolism ; physiology ; Growth Differentiation Factor 6 ; metabolism ; physiology ; Humans ; Liver ; metabolism ; Liver Diseases ; metabolism ; Smad Proteins ; metabolism

TGFβ/smad pathway is recognized as an important signal pathway to promote the pathogenesis of atherosclerosis (AS). Peroxisome proliferator-activated receptor γ (PPARγ) activation is considered to be important in modulating AS. Herein, we investigated the regulation of PPARγ on c-Ski, the repressor of TGFβ/smad pathway, in rat AS model and cultured vascular smooth muscle cells (VSMCs). c-Ski mRNA and protein expression were detected by real-time PCR and Western blot, respectively, in vivo and in vitro with treatment of PPARγ agonist rosiglitazone and antagonist GW9662. The proliferation and collagen secretion of VSMCs after c-Ski transfection were investigated. The underlying mechanism was further investigated by online program NUBIScan and luciferase reporter gene analysis. Results showed that both mRNA and protein expressions of c-Ski in the AS lesions was down-regulated in vivo, while in cultured VSMCs, c-Ski transfection significantly suppressed the proliferation and collagen secretion of rat VSMCs. Rosiglitazone significantly up-regulated mRNA and protein levels of c-Ski in VSMCs, which could be blocked by GW9662. Online NUBIScan analysis suggested possible PPARγ binding sites in the promoter region of c-Ski. In addition, luciferase activity of c-Ski reporter gene was also increased obviously in the presence of rosiglitazone. These results indicate that c-Ski is one of the newly found target genes of PPARγ and thus involved in the anti-AS effect of PPARγ.
Anilides ; pharmacology ; Animals ; Atherosclerosis ; physiopathology ; Cells, Cultured ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; PPAR gamma ; agonists ; antagonists & inhibitors ; physiology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Repressor Proteins ; genetics ; metabolism ; Signal Transduction ; Smad Proteins ; metabolism ; Thiazolidinediones ; pharmacology ; Transforming Growth Factor beta ; metabolism ; Up-Regulation

OBJECTIVETo investigate if the interaction between TGF-beta1/Smad pathway and ERK pathway in vascular smooth muscle cells exists.

METHODSThe rat arota was removed. The primary VSMC were isolated and cultured in vitro, then the VSMC were divided into four groups: (1) control group, (2) (TGF-beta1 group, (3) ERK blocking agent group, (4) TGF-beta1 + ERK blocking agent group. The expression of Smad2/3, ERK1/2 proteins, the content of phosphorylated ERK1/2 and Smad2/3 proteins were detected by Western blot, and the expression of Smad2/3 mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR) .

RESULTS(1) In contrast to control group, the content of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 group was increased (P < 0.05), that in ERK blocking agent group was decreased (P < 0.05). There was no difference between control group and TGF-beta1 + ERK blocking agent group. Compared with TGF-beta1 group, the contents of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 + ERK blocking agent group was decreased (P < 0.05). There was no difference in the expression of Smad2/3 and ERK1/2 proteins among different groups. (2) There were no differences in expression of Smad2 and Smad3 mRNA among different groups.

CONCLUSION(1) TGF-beta1 can induce Smad2/3 proteins to be phosphorylated dependent on the activated ERK pathway. (2) ERK pathway does not effect the expression of Smad2/3 at the level of protein and mRNA.

Animals ; Aorta ; cytology ; Cells, Cultured ; Female ; MAP Kinase Signaling System ; physiology ; Male ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; physiology ; Myocytes, Smooth Muscle ; physiology ; Phosphorylation ; Rats ; Rats, Wistar ; Signal Transduction ; Smad Proteins ; metabolism ; physiology ; Transforming Growth Factor beta1 ; physiology