Investigation of rs1800469 and rs1800468 Polymorphisms of the TGF-β 1 Gene in Women with Pre-eclampsia / Investigação dos Polimorfismos rs1800469 e rs1800468 do Gene TGF-β 1 em Mulheres com Pré-eclâmpsia

Abstract Objectives: to verify the contribution of polymorphisms rs1800469 and rs1800468 of the TGF-β1 gene and the risk factors for the pre-eclampsia development. Methods: this is a case-control study with 257 women from the Uberaba region of Minas Gerais were selected, 88 of them were in the pre-eclampsia group and 169 in the control group. Genotyping was performed by allelic discrimination using the real-time PCR technique. The odds ratio and the 95% confidence interval were used to evaluate the probability of the polymorphisms studied contributing for the pre-eclampsia development. The logistic regression analysis was performed to evaluate the relation among family recurrence, smoking, primiparity and the presence of polymorphic alleles and susceptibility of preeclampsia. Results: no association was found between polymorphisms rs1800469 and rs1800468 of the TGF-β1 gene and pre-eclampsia. The logistic regression analysis was statistically significant for family recurrence, showing that women with a family history of pre-eclampsia and primiparity are at an increased risk of developing the disease. Conclusions: no association was found between polymorphisms rs1800469 and rs1800468 of the TGF-β1 and pre-eclampsia gene. Factors such as family history and primiparity were associated to the risk of developing pre-eclampsia.

Resumo Objetivos: verificar a contribuição dos polimorfismos rs1800469 e rs1800468 do gene TGF-β1 e de fatores de risco para o desenvolvimento de PE. Métodos: trata-se de um estudo caso-controle, onde foram selecionadas 257 mulheres, da região de Uberaba, Minas Gerais, sendo 88 do grupo PE e 169 controles. A genotipagem foi realizada por discriminação alélica através da técnica de PCR em tempo real. Odds ratio e o intervalo de confiança de 95% foram usados para avaliar a probabilidade dos polimorfismos estudados contribuírem com o desenvolvimento de PE. A análise de regressão logística foi realizada para avaliar a relação entre recorrência familiar, tabagismo, primiparidade e presença dos alelos polimórficos e a suscetibilidade a PE. Resultados: não foi observada associação entre os polimorfismos rs1800469 e rs1800468 do gene TGF-β1 e PE. A análise de regressão logística foi estatisticamente significativa para recorrência familiar, mostrando que mulheres com histórico familiar de PE e mulheres primigestas possuem risco aumentado de desenvolver a doença. Conclusões: não foi encontrada associação entre os polimorfismos rs1800469 e rs1800468 do gene TGF-β1 e PE. Fatores como histórico familiar e primiparidade foram associados com o risco de desenvolvimento de PE.

TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes

We investigated the effect of transforming growth factor beta 1 (TGF-β1) on equine hyaluronan synthase 2 (HAS2) gene expression and hyaluronan (HA) synthesis in culture models of articular chondrocytes. Equine chondrocytes were treated with TGF-β1 at different concentrations and times in monolayer cultures. In three-dimensional cultures, chondrocyte-seeded gelatin scaffolds were cultured in chondrogenic media containing 10 ng/mL of TGF-β1. The amounts of HA in conditioned media and in scaffolds were determined by enzyme-linked immunosorbent assays. HAS2 mRNA expression was analyzed by semi-quantitative reverse transcription polymerase chain reaction. The uronic acid content and DNA content of the scaffolds were measured by using colorimetric and Hoechst 33258 assays, respectively. Cell proliferation was evaluated by using the alamarBlue assay. Scanning electron microscopy (SEM), histology, and immunohistochemistry were used for microscopic analysis of the samples. The upregulation of HAS2 mRNA levels by TGF-β1 stimulation was dose and time dependent. TGF-β1 was shown to enhance HA and uronic acid content in the scaffolds. Cell proliferation and DNA content were significantly lower in TGF-β1 treatments. SEM and histological results revealed the formation of a cartilaginous-like extracellular matrix in the TGF-β1-treated scaffolds. Together, our results suggest that TGF-β1 has a stimulatory effect on equine chondrocytes, enhancing HA synthesis and promoting cartilage matrix generation.

TGF-ß1 and its association with clinicopathological features, proliferative activity and prognosis in oral squamous cell carcinoma: an immunohistochemical study

Introduction: The prognostic value of transforming growth factor beta-1 (TGF-ß1) in oral cancer remains unclear. Therefore, the aim of this study was to evaluate TGF-ß1 expression in oral squamous cell carcinoma (OSCC) samples and its association with clinicopathological data, tumor proliferative activity, and patients' prognosis. Methods: A total of 68 patients with histopathological diagnosis of OSCC were included, as well as 9 cases of normal oral mucosa for comparison purposes. The OSCC sample was categorized according to patients' outcomes in favorable prognosis (n=30) or unfavorable prognosis (n=38). Immunohistochemical staining for TGF-ß1 and Ki-67 were performed. The slides were semi-quantitatively and quantitatively evaluated for TGF-ß1 and Ki-67, respectively. Results: TGF-ß1 was significantly increased in OSCC compared to normal oral mucosa (<0.01). An inverse correlation was found between TGF-ß1 and Ki67 staining in OSCC (p=0.01). No association was found between TGF-ß1 expression and OSCC clinicopathological features, prognosis or survival. Conclusions: TGF-ß1 had no prognostic value and appears to maintain its suppressive role concerning cell proliferation. (AU)

Silencing Histone Deacetylase 7 Alleviates Transforming Growth Factor-β1-Induced Profibrotic Responses in Fibroblasts Derived from Peyronie's Plaque

PURPOSE: Epigenetic modifications, such as histone acetylation/deacetylation and DNA methylation, play a crucial role in the pathogenesis of inflammatory disorders and fibrotic diseases. The aim of this study was to study the differential gene expression of histone deacetylases (HDACs) in fibroblasts isolated from plaque tissue of Peyronie's disease (PD) or normal tunica albuginea (TA) and to examine the anti-fibrotic effect of small interfering RNA (siRNA)-mediated silencing of HDAC7 in fibroblasts derived from human PD plaque. MATERIALS AND METHODS: For differential gene expression study, we performed reverse-transcriptase polymerase chain reaction for HDAC isoforms (1–11) in fibroblasts isolated from PD plaque or normal TA. Fibroblasts isolated from PD plaque were pretreated with HDAC7 siRNA (100 pmol) and then stimulated with transforming growth factor-β1 (TGF-β1, 10 ng/mL). Protein was extracted from treated fibroblasts for Western blotting. We also performed immunocytochemistry to detect the expression of extracellular matrix proteins and to examine the effect of HDAC2 siRNA on the TGF-β1-induced nuclear translocation of Smad2/3 and myofibroblastic differentiation. RESULTS: The mRNA expression of HDAC2, 3, 4, 5, 7, 8, 10, and 11 was higher in fibroblasts isolated from PD plaque than in fibroblasts isolated from normal TA tissue. Knockdown of HDAC7 in PD fibroblasts inhibited TGF-β1-induced nuclear shuttle of Smad2 and Smad3, transdifferentiation of fibroblasts into myofibroblasts, and abrogated TGF-β1-induced production of extracellular matrix protein. CONCLUSIONS: These findings suggest that specific inhibition of HDAC7 with RNA interference may represent a promising epigenetic therapy for PD.

Effects of ginsenoside Re on vascular intimal proliferation in balloon-injuried rats via regulation of TGF-β1/Smads signaling pathway / 中草药

Objective: To investigate the inhibitory effects of ginsenoside Re on intimal hyperplasia in balloon-injuried rats and further explore the role of TGF-β1/Smads signaling pathway in this protection. Methods: Fifty SD rats were randomly divided into five groups, including Sham operation group, model group, ginsenoside Re low, medium, and high-dose groups. The injured model of carotid artery intima was established by 2F balloon catheters in each group except the sham operation group. One day after model was established, animals were daily ig administered with distilled water in model group, Sham operation group, and ginsenoside Re (12.5 mg/kg, 25 mg/kg and 50 mg/kg) groups. Two weeks later, animals were sacrificed and the injured artery was taken for HE staining. The histopathological changes were observed and the lumen area, intima area, and media area as well as the ratio of intimal area/media area were detected. The expression of transforming growth factors β1 (TGF-β1), SMAD family member 2 (Smad 2) and Smad family member 3 (Smad 3) were measured by real time RT-PCR and immunohistochemistry. Results: Compared with the Sham operation group, the vessel cavity in the model group was narrower (P < 0.01); Compared with the model group, the medium and high dose of ginsenoside Re obviously alleviated vascular intimal hyperplasia (P < 0.05). Compared with the Sham operation group, the mRNA and protein expressions levels of TGF-β1, Smad 2, and Smad 3 in model group were higher (P < 0.01), which were obviously decreased in the medium and high-dose ginsenoside Re (P < 0.05). Conclusion: Ginsenoside Re could alleviate the vascular neointimal hyperplasia through suppressing the TGF-β1/Smads signaling pathway.

Effect of acupuncture plus thunder-fire moxibustion on MMP-3, TIMP-1 and TGF-β1 in rats with knee osteoarthritis / 针灸推拿医学（英文版）

Objective: To observe the effect of acupuncture plus thunder-fire moxibustion on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in cartilage of knee osteoarthritis (KOA) rats, and to explore the mechanism of acupuncture plus thunder-fire moxibustion in the treatment of KOA. Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a blank control group, a model group and an acupuncture-moxibustion group by random digits table, 10 rats in each group. Rats in the model group and the acupuncture-moxibustion group were injected with papain in the right posterior knee joint to prepare the models. The levels of MMP-3 and TIMP-1 in rat synovium of each group were measured by enzyme-linked immunosorbent assay (ELISA) after 2 weeks of treatment. The level of TGF-β1 was determined by Motic B5 Micro-camera system. Results:The levels of MMP-3 and TIMP-1 in the cartilage of the model group were significantly higher than those in the blank control group (allP<0.01); the levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were lower than those in the model group, and the between-group differences were statistically significant (all P<0.05). The levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were higher than those in the blank control group, and the differences were statistically significant (all P<0.05). The level of TGF-β1 in cartilage tissues of the model group was significantly lower than that in the blank control group (P<0.01); the level of TGF-β1 in the acupuncture-moxibustion group was higher than that in the model group (P<0.05), but it was lower than that in the blank control group, and the between-group difference was statistically significant (P<0.05). Conclusion: Acupuncture plus thunder-fire moxibustion can effectively recover the abnormal expressions of MMP-3 and TIMP-1 in KOA model rats and somewhat up-regulate TGF-β1, which may be one of its mechanisms of acupuncture plus thunder-fire for KOA.

Effect of acupuncture plus thunder-fire moxibustion on MMP-3, TIMP-1 and TGF-β1 in rats with knee osteoarthritis / 针灸推拿医学（英文版）

Objective: To observe the effect of acupuncture plus thunder-fire moxibustion on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in cartilage of knee osteoarthritis (KOA) rats, and to explore the mechanism of acupuncture plus thunder-fire moxibustion in the treatment of KOA. Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a blank control group, a model group and an acupuncture-moxibustion group by random digits table, 10 rats in each group. Rats in the model group and the acupuncture-moxibustion group were injected with papain in the right posterior knee joint to prepare the models. The levels of MMP-3 and TIMP-1 in rat synovium of each group were measured by enzyme-linked immunosorbent assay (ELISA) after 2 weeks of treatment. The level of TGF-β1 was determined by Motic B5 Micro-camera system. Results:The levels of MMP-3 and TIMP-1 in the cartilage of the model group were significantly higher than those in the blank control group (allP<0.01); the levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were lower than those in the model group, and the between-group differences were statistically significant (all P<0.05). The levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were higher than those in the blank control group, and the differences were statistically significant (all P<0.05). The level of TGF-β1 in cartilage tissues of the model group was significantly lower than that in the blank control group (P<0.01); the level of TGF-β1 in the acupuncture-moxibustion group was higher than that in the model group (P<0.05), but it was lower than that in the blank control group, and the between-group difference was statistically significant (P<0.05). Conclusion: Acupuncture plus thunder-fire moxibustion can effectively recover the abnormal expressions of MMP-3 and TIMP-1 in KOA model rats and somewhat up-regulate TGF-β1, which may be one of its mechanisms of acupuncture plus thunder-fire for KOA.

Retinoid X receptor agonist inhibits TGF-β1-induced collagen synthesis in cardiac fibroblasts by repressing Smad2 activation / 中国病理生理杂志

AIM:To investigate the effect of activation of retinoid X receptor (RXR) on transforming growth factor β1 (TGF-β1) induced collagen synthesis under hypoxic environment in rat cardiac fibroblasts (CFs) and underlying molecular mechanisms .METHODS: CFs were cultured using myocardial tissue with dry method .Hypoxic environment was established for CFs by continuous nitrogen supplement .Type I and type III collagens in supernatants were detected by ELISA.Nuclear and cytoplasmic extractions were prepared using NE-PER nuclear and cytoplasmic extraction reagents .The protein levels of Smad2 and p-Smad2 were determined by Western blot and immunocytochemical staining .RESULTS:Un-der hypoxic condition , TGF-β1 (0.01~10 μg/L) increased the synthesis of type I and type III collagens in a dose-de-pendent manner in the CFs .At the concentration of 5μg/L, the synthesis of collagen I and III was significantly increased as compared with control group (P<0.01).RXR agonist 9-cis-retinoic acid (9-cis-RA;10 -9 ~10 -6 mol/L) decreased TGF-β1 (5μg/L)-induced synthesis of type I and III collagens in a dose-dependent manner in the CFs under hypoxic con-dition.The synthesis of type I and type III collagens was significantly inhibited by 9-cis-RA (P<0.01).Smad2 inhibitor ( 20 nmol/L) showed similar inhibitory effect on the synthesis of type I and III collagens induced by TGF -β1 under hypoxic condition.Compared with TGF-β1 intervention group, the cytoplasmic level of p-Smad2 in the CFs was significantly in-creased in TGF-β1+9-cis-RA group, but the nuclear p-Smad2 level was significantly decreased (P<0.05).CONCLU-SION:Retinoid X receptor agonist 9-cis-RA inhibits TGF-β1-induced synthesis of type I and type III collagens in the CFs by repressing p-Smad2 nuclear translocation under hypoxic condition .

Loss of IκB kinase β promotes myofibroblast transformation and senescence through activation of the ROS-TGFβ autocrine loop

Using forward and reverse genetics and global gene expression analyses, we explored the crosstalk between the IκB kinase β (IKKβ) and the transforming growth factor β (TGFβ) signaling pathways. We show that in vitro ablation of Ikkβ in fibroblasts led to progressive ROS accumulation and TGFβ activation, and ultimately accelerated cell migration, fibroblast-myofibroblast transformation and senescence. Mechanistically, the basal IKKβ activity was required for anti-oxidant gene expression and redox homeostasis. Lacking this activity, IKKβ-null cells showed ROS accumulation and activation of stress-sensitive transcription factor AP-1/c-Jun. AP-1/c-Jun activation led to up-regulation of the Tgfβ2 promoter, which in turn further potentiated intracellular ROS through the induction of NADPH oxidase (NOX). These data suggest that by blocking the autocrine amplification of a ROS-TGFβ loop IKKβ plays a crucial role in the prevention of fibroblast-myofibroblast transformation and senescence.

Immunohistochemical expression of TGF-ß1 and osteonectin in engineered and Ca(OH)2-repaired human pulp tissues

Abstract The aim of the present study was to evaluate the expression of transforming growth factor-β1 (TGF-β1) and osteonectin (ON) in pulp-like tissues developed by tissue engineering and to compare it with the expression of these proteins in pulps treated with Ca(OH)2 therapy. Tooth slices were obtained from non-carious human third molars under sterile procedures. The residual periodontal and pulp soft tissues were removed. Empty pulp spaces of the tooth slice were filled with sodium chloride particles (250-425 µm). PLLA solubilized in 5% chloroform was applied over the salt particles. The tooth slice/scaffold (TS/S) set was stored overnight and then rinsed thoroughly to wash out the salt. Scaffolds were previously sterilized with ethanol (100-70°) and washed with phosphate-buffered saline (PBS). TS/S was treated with 10% EDTA and seeded with dental pulp stem cells (DPSC). Then, TS/S was implanted into the dorsum of immunodeficient mice for 28 days. Human third molars previously treated with Ca(OH)2 for 90 days were also evaluated. Samples were prepared and submitted to histological and immunohistochemical (with anti-TGF-β1, 1:100 and anti-ON, 1:350) analyses. After 28 days, TS/S showed morphological characteristics similar to those observed in dental pulp treated with Ca(OH)2. Ca(OH)2-treated pulps showed the usual repaired pulp characteristics. In TS/S, newly formed tissues and pre-dentin was colored, which elucidated the expression of TGF-β1 and ON. Immunohistochemistry staining of Ca(OH)2-treated pulps showed the same expression patterns. The extracellular matrix displayed a fibrillar pattern under both conditions. Regenerative events in the pulp seem to follow a similar pattern of TGF-β1 and ON expression as the repair processes.

Effect of Zuoguiwan and Youguiwan on transforming growth factor beta1 and its signal transduction protein Smad2/3 in osteogenic induction of rat bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE:To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factorβ1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cellcultivation was processed in five groups:bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, andβ-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I col agen. The immunohistochemical assay was utilized to test transforming growth factorβ1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION:It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I col agen, transforming growth factorβ1 and Smad2/3 expression, compared with blank serum group and inducer group (P<0.05);moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P<0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells.

Effects of Total Flavonoids from Portulaca on Transforming Growth Factor β1 in Rats with Hepatic Fibrosis / 医药导报

Objective To explore the effects of total flavonoids from portulaca against liver fibrosis in rats by detecting TGF-β1 gene and protein expressions. Methods A total of 48 SD rats were randomly divided into normal control, model control, glucyrrhizin aqueous,and total flavonoids groups,with 12 rats in each group. Except those in the normal control group, rats in other groups were intraperitoneally injected with 2 mL · kg-1 · d-1 carbon tetrachloride to induce liver fibrosis. Rats in glucyrrhizin aqueous group and total flavonoids ones were intragastrically administered with 15. 75 mg · kg-1 of glycyrrhizin aqueous solution or 35. 6 mg·kg-1 of total flavonoids aqueous solution,respectively. The normal and model control groups were administered with equal volume of aqueous solution. Thirty days later,rats were sacrificed by anesthesia. Livers were obtained to detect TGF-β1 gene and protein expressions by RT-PCR and Western-Blot. Results Relative gene expression of TGF-β1 in the normal control,model control,glucyrrhizin aqueous and flavonoids groups was 0. 725±0. 130,7. 493±1. 410,3. 016±1. 240,and 2. 668±1. 150,respectively. Total flavonoids from portulaca significantly reduced the gene (P<0. 01) and protein (P<0. 01) expressions of TGF-β1 . Conclusion Efficacy of total flavonoids from portulaca in treating hepatic fibrosis may be related to decreased TGF-β1 expression in rats.

Platelet-Rich Plasma: Quantitative Assessment of Growth Factor Levels and Comparative Analysis of Activated and Inactivated Groups

BACKGROUND: Platelet-rich plasma (PRP) has more concentrated platelets than normal plasma (approximately 150-400x10(3) cell/dL). Platelets excrete several growth factors and cytokines that are associated with the healing and regeneration process. However, even though PRP is widely used, the mechanism or actual effect is presently unclear. Therefore, this study was performed to investigate the levels of growth factors and platelet concentration rate. METHODS: Autologous blood for preparing PRP was obtained from healthy subjects aged 25 to 35 years. The samples were divided into 4 experimental groups (inactivated whole blood, inactivated PRP, activated whole blood with thrombin and calcium chloride, and activated PRP). The platelet counts in the blood were analyzed and the growth factors were quantitatively measured. A statistical analysis was performed by using Dunn's multiple comparison test. RESULTS: In the blood cell analysis, the platelet count of the PRP group was approximately 4.25 times higher than that of the whole blood group. In the quantitative analysis of growth factors, the platelet-derived growth factor (PDGF)-AB, PDGF-BB, and transforming growth factor-beta of the inactivated and activated PRP groups were higher than those of the inactivated and activated whole blood groups (P<0.05). CONCLUSIONS: In this study, the platelet count and the levels of PDGF-AB and PDGF-BB in the PRP were determined. Further, more research is required on the bioactivity level of the growth factors secreted during the process of PRP preparation and the potency of growth factors that can be exerted physiologically in vivo.

The Role of Focal Adhesion Kinase in the TGF-beta-Induced Myofibroblast Transdifferentiation of Human Tenon's Fibroblasts

PURPOSE: To investigate the role of focal adhesion kinase (FAK) in transforming growth factor (TGF)-beta-induced myofibroblast transdifferentiation of human Tenon's fibroblasts. METHODS: Primary cultured human Tenon's fibroblasts were exposed to TGF-beta1 for up to 48 hours. The mRNA levels of FAK, alpha smooth muscle actin (alphaSMA), and beta-actin were determined by quantitative real time reverse transcription polymerase chain reaction. The protein levels of collagen type I, FAK, phospho-FAK, alphaSMA, and beta-actin were determined by Western immunoblots. After the small interfering RNA targeting FAK (siRNA(FAK)) molecules were delivered into the cells, the expressions of alphaSMA proteins were determined by Western immunoblots. RESULTS: In human Tenon's fibroblasts, TGF-beta1 significantly increased the mRNA and protein expressions of alphaSMA. However, when the action of FAK was inhibited using siRNAFAK, the TGF-beta1-induced expression of alphaSMA was attenuated. CONCLUSIONS: Our data suggest that FAK may be associated with the TGF-beta1-induced transdifferentiation of human Tenon's fibroblasts to myofibroblasts, which is the essential step of subconjunctival fibrosis.

Expression of decorin and transforming growth factor-β_1 mRNA and type Ⅰ collagen in the anterior vagina of women with pelvic organ prolapse / 中国医师进修杂志

Objective To determine quantity of type Ⅰ collagen and mRNA expression of decorin and transforming growth factor (TGF)- β_1 in the upper portion of the anterior vagina in women with pelvic organ prolapse (POP). Methods Transvaginat biopsies were obtained from the anterior vaginal wall in 27 POP(POP group) and 21 non-poP(control group). The concentration of type Ⅰ collagen was determined by ELISA technique. RT-qPCR was used to verify the mRNA level of decorin and TGF- β_1. Results The mean concentration of type Ⅰ collagen in the premenopausal women in POP group [(47.94 ± 6.82) mg/g total protein (TP)]was significantly lower than that in control group [(62.33±6.57) mg/g TP],in the postmenopausal women in POP group, it was (41.82±5.96) mg/g TP, significantly lower than that in control group[(61.21 ± 4.84)mg/g TP](P< 0.01). The mRNA expression of decorin was 1.61 ± 0.10 in POP group, significantly lower than that in control group 2.77±0.60 ,while that of TGF-β_1 was higher in POP greup10.86 ± 0.23 than that in control group 9.87 ± 0.92 (P < 0.05). In the postmenopausal women in two groups, there were no significant difference in the mRNA expression of decorin and TGF-β_1 (P > 0.05). Conclusions A decrease in quantity of type Ⅰ coUagen in the connective tissue of anterior vagina may compromise the tensile strength and an increase susceptibihty to prolapse. Deeorin might be involved in the pathogenesis in premenopausal women with POP. The pathogenesis of POP is muhifactorial.

Bone Morphogenetic Protein Signaling: Implications in Urology

The bone morphogenetic proteins (BMPs), as members of the transforming growth factor-beta (TGF-beta) superfamily, not only control bone formation, but also regulate multiple key steps during embryonic development and differentiation. Furthermore, BMPs play critical roles in maintaining the homeostasis of the cardiovascular, pulmonary, reproductive, urogenital, and nervous systems in adult life. Like all members of the TGF-beta superfamily, BMP signaling is mediated through a heteromeric complex of type I and type II transmembrane serine/threonine kinase receptors. The subsequent signal transduction cascade includes either the canonical Smad-dependent or non-canonical Smad-independent pathways. Reflecting the critical function of BMPs, BMP signaling is tightly regulated at multiple steps by various mechanisms including extracellular endogenous antagonists, neutralizing antibodies/extracellular soluble receptor domains, small molecule inhibitors, cytoplasmic inhibitory Smads, and transcriptional co-repressors. Recently, dorsomorphin, the first small molecule inhibitor of BMP signaling, was identified and suggested as a useful tool for dissecting the mechanisms of signaling pathways and for developing novel therapeutics for diverse human diseases that are related to the BMP signaling pathways. In this article, we discuss various mechanisms involved in regulating BMP signaling pathways and their implications for urology.

Expression of latent transforming growth factor beta binding protein-1 and transforming growth factor beta type Ⅱ receptor in condyloma acuminatum tissues / 中华皮肤科杂志

Objective To investigate the role of the expression of latent transforming growth factor beta binding protein-1 (LTBP-1) and transforming growth factor beta receptor type Ⅱ(TGF-beta R Ⅱ) in the pathogenesis of condyloma acuminatum (CA). Methods Samples were resected from the lesions of 30 patients with CA and prepuces of 17 normal human controls. The mRNA and protein expressions of LTBP-1 and TGF-betaR Ⅱ were assessed by quantitative real-time PCR and a streptavidin-biotin-peroxidase staining technique, respectively. Results As shown by Real time PCR, the mRNA expression levels of LTBP-1 and TGF-betaR Ⅱ were significantly higher in CA tissues than those in the controls, with the average value of 2 (-Delta Delta α) being 2.46 and 3.43, respectively. A lower intensity of stainning was observed for LTBP-1 and TGF-betaR Ⅱ in CA tissues compared with the normal controls (182.51±9.89 vs 167.78±12.56, 187.35± 11.23 vs 170.15±13.21, t = 5.62, 3.70 respectively, both P <0.01). Conclusion The decrease in the expres-sion of both LTBP-1 and TGF-betaR Ⅱ may lead to the abnormality in the activation of TGF-beta and signal transduction pathways.

The Inhibitory Effect of siRNAs on The High Glucose-Induced Overexpression of TGF-beta1 in Mesangial Cells

Diabetic nephropathy is characterized by an expansion of the glomerular mesangium, caused by mesangial cell proliferation and an excessive accumulation of extracellar matrix (ECM) proteins, which eventually leading to glomerulosclerosis. TGF-beta1 was found to play an important role in the accumulation of ECM in the kidney. In this study, TGF-beta1 RNA interference was used as an effective therapeutic strategy. The inhibitory effect of TGF-beta1 small interfering RNAs (siRNAs) on the high glucose-induced overexpression of TGF-beta1 in rat mesangial ceys (RMCs). A high levels of glucose induces TGF-beta1 mRNA and protein, and TGF-beta1 siRNAs reduce the ability of high glucose to stimulate their expression. We also examined the inhibitory effect of TGF-beta1 siRNAs on the expression of plasminogen activator inhibitor (PAI)-1 and Collagen Type I which are down-regulators of TGF-beta1. The expression of TGF-beta1, PAI-1 and Collagen Type I was increased in RMCs that were stimulated by 30 mM glucose. TGF-beta1 siRNAs reduces high glucose-induced TGF-beta1, PAI-1, and Collagen Type I mRNA and protein expression in a dose-dependent manner. In conclusion, the present study demonstrates that TGF-beta1 siRNAs effectively inhibits TGF-beta1 mRNA and protein expression in RMCs. These suggest that TGF-beta1 siRNAs through RNAi may be a useful tool for developing new therapeutic applications for the treatment of diabetic nephropathy.

Effect of Antisense TGF-beta1 Oligodeoxynucleotides in Streptozotocin-Induced Diabetic Rat Kidney

Transforming growth factor (TGF)-beta1 is an important fibrogenic factor that is involved in the pathogenesis of diabetic nephropathy. We evaluated the effect of circular antisense TGF-beta1 oligodeoxynucleotides (ODNs) on the TGF-beta1 expression in the rat mesangial cell culture and in streptozotocin (STZ)-induced diabetic rats. Circular antisense TGF-beta1 ODNs were found to be stable in rat serum, significantly decreasing TGF-beta1 mRNA expression compared with linear antisense ODNs in the rat mesangial cell culture. Circular antisense TGF-beta1 ODNs were introduced into the tail vein of normal rats using hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer method and were confirmed to be delivered effectively into the kidney, liver, lungs, and spleen. To inhibit the overexpression of TGF-beta1 in diabetic kidneys, we introduced circular antisense TGF-beta1 ODNs into the STZ-induced diabetic rats. On day 13 after circular antisense TGF-beta1 ODNs injection, TGF-beta1 mRNA and protein expression markedly decreased and urinary TGF-beta1 excretion rate also dropped in the circular antisense TGF-beta1 ODNs-treated diabetic rats. These results suggest that circular antisense TGF-beta1 ODNs may be a useful tool for developing new therapeutic application for progressive diabetic nephropathy.

Effect of bone morphogenetic protein-7 on transforming growth factor ? signaling in hepatic stellate cells / 第三军医大学学报

Objective To observe the effect of bone morphogenetic protein-7 (BMP-7) on proliferation,activation and TGF? signaling in TGF?1 inducing hepatic stellate cells (HSC),and its anti-fibrosis mechanism.Methods Human HSC LX-2 cell line was treated with BMP-7 at different concentrations (80,40,20 ng/ml) and TGF?1(5 ng/ml).Proliferation of HSC LX-2 cells was detected with cell counting kit-8 (CCK8).Expressions of ?-smooth muscle actin (?-SMA) and collagen Ⅰ,as well as TGF receptors Ⅰ and Ⅱ (TGF?RⅠ,TGF?RⅡ) mRNA,Smad 3,7 mRNAs,were detected by immunocytochemical assay and RT-PCR,respectively.Results No significant difference was found in proliferation of LX-2 cells before and after treatment with TGF?1.BMP-7 used at different concentrations (80,40,20 ng/ml) inhibited the proliferation of LX-2 with an inhibition rate of 28.9%,19.6% and 10.5%,respectively (P