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1.
Arch. endocrinol. metab. (Online) ; 65(2): 172-184, Mar.-Apr. 2021. tab, graf
Article in English | LILACS | ID: biblio-1248817

ABSTRACT

ABSTRACT Objective: Abnormalities involving the TGFB1 gene and its receptors are common in several types of cancer and often related to tumor progression. We investigated the role of single nucleotide polymorphisms (SNP) in the susceptibility to cancer, their impact on its features, as well as the role of mRNA expression of these genes in thyroid malignancy. Materials and methods: We genotyped TGFB1, TGFBR1, and TGFBR2 SNPs in 157 papillary thyroid cancer (PTC) patients and 200 healthy controls. Further, we investigated RNA samples of 47 PTC and 80 benign nodules, searching for differential mRNA expression. Results: SNPs rs1800472 and rs1800469 were associated with characteristics of PTC aggressiveness. Effect predictor software analysis of nonsynonymous SNP rs1800472 indicated increasing protein stability and post-translational changes. TGFB1 mRNA expression was upregulated in PTC and downregulated in benign samples, differentiating malignant from benign nodules (p<0.0001); PTC from goiter (p<0.0001); and PTC from FA (p<0.0001). TGFBR1 mRNA expression was upregulated in goiter and PTC, but downregulated in FA, distinguishing PTC from goiter (p=0.0049); PTC from FA (p<0.0001); and goiter from FA (p=0.0267). On the other hand, TGFBR2 was downregulated in all histological types analyzed and was not able to differentiate thyroid nodules. Conclusion: TGFB1 polymorphism rs1800472 may confer greater activity to TGF-β1 in the tumor microenvironment, favoring PTC aggressiveness. Evaluation of TGFB1 and TGFBR1 mRNA levels may be useful to identify malignancy in thyroid nodules.


Subject(s)
Thyroid Nodule/genetics , Transforming Growth Factor beta1/genetics , Receptor, Transforming Growth Factor-beta Type I/genetics , Receptor, Transforming Growth Factor-beta Type II/genetics , RNA, Messenger/genetics , Thyroid Neoplasms , Polymorphism, Single Nucleotide , Tumor Microenvironment
2.
Chinese Journal of Traumatology ; (6): 360-367, 2021.
Article in English | WPRIM | ID: wpr-922355

ABSTRACT

PURPOSE@#To observe the changes of gait behavior and the expression of wound healing factors of transforming growth factor-β1 (TGF-β1), TGF-β3 and cAMP response element binding protein-1 (CREB-1) during the healing of Achilles tendon in a rat model, and to investigate whether gait analysis can be used to evaluate the tendon healing.@*METHODS@#Achilles tendon of 40 healthy male Sprague-Dawley rats were transected and sutured to establish the Achilles tendon injury (ATI) model. They were randomly divided into 4 groups based on the observational time point at 1, 2, 4 and 6 weeks after injury (n = 10 for each group). Before modeling, 9 rats were randomly selected for CatWalk gait analysis, which contained step cycle, single stance time and average speed. Data were recorded as the normal controls. After then, ATI models were established in the left hind limbs of the all 40 rats (ATI group), while the right hind limbs were only cut and sutured without injury of the Achilles tendon (sham operation group). At 1, 2, 4 and 6 weeks after injury, the gait behavior of the corresponding group of rats (n = 9) as observed and recorded by CatWalk platform. After then, the rats were sacrificed and Achilles tendon of both limbs was harvested. The tendon healing was observed by gross anatomy and histological examination, and the protein and mRNA expression of TGF-β1, TGF-β3, CREB-1 were observed by immunohistochemistry and qPCR. The results of tendon gross grading were analyzed by Wilcoxon rank sum test, and other data were analyzed by one-way analysis of variance among multiple groups.@*RESULTS@#Compared with normal controls, all gait indexes (step cycle, single stance time and average speed) were greatly affected following ATI, which however improved with time. The step cycle was significantly lower at 1, 2 and 4 weeks after ATI (compared with normal controls, all p  0.05). The single stance time of the ATI group was significantly shorter at 1 and 2 weeks after operation ((0.078 ± 0.010) s at 1 week, (0.078 ± 0.020) s at 2 weeks, all p < 0.001) and revealed no significant difference at 4 weeks (p = 0.120). The average speed of ATI group at 1, 2, 4, 6 weeks was significantly lower than that in the normal control group (all p < 0.001). Gross observation showed that the grade of local scar adhesion in ATI group increased significantly at 2, 4 and 6 weeks, compared with the sham operation group (all p < 0.001). Extensive adhesion was formed at 6 weeks after ATI. The results of HE staining showed that the number of fibroblast increased gradually and arranged more orderly in ATI group at 1, 2 and 4 weeks (all p < 0.001), and decreased at 6 weeks, but it was still significantly higher than that of the sham operation group (p < 0.001). Immunohistochemistry showed that the positive expression of TGF-β1, TGF-β3, CREB-1 in ATI group was higher than that in the sham operation group at 4 time points (all p < 0.05), which reached the peak at 2 weeks after operation and decreased at 4 weeks (p = 0.002, p < 0.001, p = 0.041, respectively). The results of qPCR suggested that the mRNA expression of TGF-β1, TGF-β3, CREB-1 in ATI group was higher than that in the sham operation group at all-time points (all p < 0.05), which reached the peak at 2 weeks after operation, decreased at 4 weeks, and significantly decreased at 6 weeks (all p < 0.001).@*CONCLUSION@#Gait behavior indexes are associated with Achilles tendon healing. The study gives an insight of TGF-β1, TGF-β3, CREB-1 changes in the coursing of Achilles tendon healing and these cytokines may be able to be used to regulate the Achilles tendon healing.


Subject(s)
Achilles Tendon , Animals , CREB-Binding Protein , Gait Analysis , Male , Rats , Rats, Sprague-Dawley , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta3 , Wound Healing
3.
Article in Chinese | WPRIM | ID: wpr-878923

ABSTRACT

This study aimed to assess whether chrysin(ChR) can inhibit epithelial-mesenchymal transition(EMT) of type Ⅱ alveolar epithelial cell and produce anti-pulmonary fibrosis effect by regulating the NF-κB/Twist 1 signaling pathway. Sixty rats were randomly divided into the control group, the bleomycin(BLC) group, BLC+ChR(50 mg·kg~(-1)) group and BLC+ChR(100 mg·kg~(-1)) group, with 15 rats in each group. The pulmonary fibrosis model was induced by intratracheal injection of BLC(7 500 U·kg~(-1)). Rats were orally administered with different doses of ChR after BLC injection for 28 days. The cells were divided into control group, TGF-β1 group(5 ng·mL~(-1)), and TGF-β1+ChR(1, 10, 100 μmol·L~(-1)) groups. The type Ⅱ alveolar epithelial cells were treated with TGF-β1 for 24 h, and then treated with TGF-β1 for 48 h in the presence or absence of different doses of ChR(1, 10 and 100 μmol·L~(-1)). The morphological changes and collagen deposition in lung tissues were analyzed by HE staining, Masson staining and immunohistochemistry. The mRNA and protein expression levels of collagen Ⅰ, E-cadherin, zonula occludens-1(ZO-1), vimentin, alpha smooth muscle actin(α-SMA), inhibitor of nuclear factor kappa B alpha(IκBα), nuclear factor-kappa B p65(NF-κB p65), phospho-NF-κB p65(p-p65) and Twist 1 in lung tissues and cells were detected by qPCR and Western blot, respectively. The animal experiment results showed that as compared with the BLC group, after administration of ChR for 28 days, bleomycin-induced pulmonary fibrosis in rats was significantly relieved, collagen Ⅰ expression in lung tissues was significantly reduced(P<0.05 or P<0.01), and EMT of alveolar epithelial cells was obviously inhibited [the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], concomitantly with significantly reduced IκBα and p65 phosphorylation level in cytoplasm and decreased NF-κB p65 and Twist 1 expression in nucleus(P<0.05 or P<0.01). The cell experiment results showed that different doses of ChR(1, 10 and 100 μmol·L~(-1)) significantly reduced TGF-β1-induced collagen Ⅰ expression(P<0.05 or P<0.01), significantly inhibited EMT of type Ⅱ alveolar epithelial cells[the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], and inhibited IκBα and p65 phosphorylation in cytoplasm and down-regulated NF-κB p65 and Twist 1 expression in nucleus induced by TGF-β1(P<0.05 or P<0.01). The results suggest that ChR can reverse EMT of type Ⅱ alveolar epithelial cell and alleviate pulmonary fibrosis in rats, and its mechanism may be associated with reducing IκBα phosphorylation and inhibiting NF-κB p65 phosphorylation and nuclear transfer, thus down-regulating Twist 1 expression.


Subject(s)
Alveolar Epithelial Cells/metabolism , Animals , Epithelial-Mesenchymal Transition , Flavonoids , NF-kappa B/metabolism , Rats , Signal Transduction , Transforming Growth Factor beta1/genetics
4.
Article in Chinese | WPRIM | ID: wpr-880757

ABSTRACT

OBJECTIVE@#To investigate high-salt exposure-induced polarization of mononuclear macrophages and the changes in proliferation and phenotypic transformation of renal fibroblasts in a co-culture system.@*METHODS@#Cultured mononuclear macrophages were exposed to high salt (161 mmol/L Na +) for 2 h and the surface markers of M0, M1 and M2-type macrophages were detected with RT-qPCR. The culture medium of the macrophages in normal and high-salt groups was collected for detection of the mRNA and protein levels of IL-6 and TGF-β1 using RT-qPCR and ELISA. A co-culture system of high salt-exposed macrophages and renal fibroblasts (NRK-49F) was established using a Transwell chamber, and the changes in proliferation and migration of NRK-49F cells were examined using EdU assay and Transwell assay, respectively. Western blotting was performed to detect the expressions of collagen I, collagen III and collagen α-SMA in NRK-49F cells.@*RESULTS@#The high salt-exposed macrophages showed significantly increased mRNA levels of M2-type macrophage surface markers mannose receptor and arginase (@*CONCLUSIONS@#High-salt exposure induces polarization of mononuclear macrophages into M2-type macrophages and promotes secretion of IL-6 and TGF-β1 by the macrophages to induce the proliferation and phenotypic transformation of NRK-49F cells.


Subject(s)
Cell Proliferation , Coculture Techniques , Fibroblasts , Kidney , Macrophages , Transforming Growth Factor beta1/genetics
5.
Braz. j. med. biol. res ; 52(10): e8324, 2019. tab, graf
Article in English | LILACS | ID: biblio-1039243

ABSTRACT

The aim of this study was to investigate the role of kinase-insert domain-containing receptor (KDR) in intrauterine adhesions (IUA) and its mechanism. The Case group consisted of 92 patients diagnosed with IUA, and the Control group included 86 patients with uterine septum who had normal endometrium verified with an uteroscope. In addition, 50 rats were randomly assigned into Control, Sham, Model, NC-siRNA, and KDR-siRNA groups. Rats in the Model, NC-siRNA, and KDR-siRNA groups were induced by uterine curettage and lipopolysaccharide (LPS) treatment to establish the IUA model. Then, immunohistochemistry was applied for detection of VEGF and KDR expression, HE staining was used for observation of the endometrial morphology and gland counting, Masson staining for measurement of the degree of endometrial fibrosis, and qRT-PCR and western blot for the expression of KDR, VEGF, MMP-9, as well as TGF-β1/Smads pathway-related proteins. Compared with the Control group, the mRNA and protein expressions of KDR were significantly higher in IUA endometrial tissues, and the expression of KDR was positively correlated to the severity of IUA. In addition, the injection of si-KDR increased the number of endometrial glands, reduced the area of fibrosis, inhibited mRNA and protein expression of KDR and VEGF, up-regulated the expression of MMP-9 and Smad7, and decreased the expression level of TGF-β1, p-Smad2, p-Smad3, and Smad4 in rats with IUA. Highly-expressed KDR was related to patients' severity of IUA, and silencing KDR may prevent the occurrence and development of IUA via TGF-β1/Smads signaling pathway and up-regulating the expression of MMP-9.


Subject(s)
Humans , Animals , Female , Adult , Middle Aged , Rats , Young Adult , Uterine Diseases/metabolism , Signal Transduction , Tissue Adhesions/metabolism , Vascular Endothelial Growth Factor Receptor-2/metabolism , Transforming Growth Factor beta1/metabolism , Uterine Diseases/pathology , Severity of Illness Index , Immunohistochemistry , Case-Control Studies , Tissue Adhesions/pathology , Blotting, Western , Rats, Wistar , Vascular Endothelial Growth Factor Receptor-2/genetics , Disease Models, Animal , Smad Proteins/genetics , Smad Proteins/metabolism , Transforming Growth Factor beta1/genetics , Real-Time Polymerase Chain Reaction
6.
Rev. Soc. Bras. Med. Trop ; 50(3): 301-308, May-June 2017. tab
Article in English | LILACS | ID: biblio-896975

ABSTRACT

Abstract INTRODUCTION: Transforming growth factor-beta 1 (TGFβ1) is a potent suppressive cytokine that contributes to chronic hepatitis B (CHB) infection. Disparities in TGFβ1 production among individuals have been attributed to TGFβ1 genetic polymorphisms. We examined whether three putative polymorphisms in TGFβ1[-509 C/T (rs1800469), +869 C/T (rs1800470), and +11929 C/T (rs1800472)]are associated with CHB infection in a South-Eastern Iranian population. METHODS: In total, 341 subjects were recruited, including 178 patients with CHB and 163 healthy individuals as controls. Genotyping of the three TGFβ1 SNPs was performed by tetra amplification refractory mutation system-PCR. RESULTS: TheTGFβ1 +869 TT vs.CC genotype in codominant (OR=0.445, p=0.012) and TT vs. TC+CC in the recessive (OR=0.439, p=0.003) model as well as the variant allele T vs. C(OR=0.714, p=0.038) were associated with lower CHB infection risk. However, the +11929 C/T polymorphism was associated with increased CHB risk, and the CT vs. CC genotype (OR=2.77, P=0.001) and T variant allele (OR=2.53, P=0.002) were risk factors for CHB. Furthermore, TTT (+869/-509/+11929) and CCC haplotypes were risk and protective factors for CHB, respectively. We found no significant association between viral DNA load and TGFβ1 genotype or hepatic enzyme levels (p >0.05). CONCLUSIONS: Results indicated that the TGFβ1+869TT genotype and T allele were protective factors, whereas the +11929 CT genotype and T allele were risk factors for CHB infection.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Aged , Young Adult , Polymorphism, Genetic , Extracellular Matrix Proteins/genetics , Transforming Growth Factor beta/genetics , Hepatitis B, Chronic/genetics , Genetic Predisposition to Disease , Transforming Growth Factor beta1/genetics , Case-Control Studies , Gene Frequency , Genotype , Iran , Middle Aged
7.
Mem. Inst. Oswaldo Cruz ; 111(11): 663-669, Nov. 2016. tab
Article in English | LILACS | ID: biblio-829247

ABSTRACT

Human papillomavirus (HPV) infections are strongly associated with the development of cervical intraepithelial neoplasias and invasive cervical cancer. Polymorphisms in cytokine-encoding genes and behavioural cofactors could play an important role in protecting an individual against viral infections and cancer. Here, we investigated whether IL-6 -174 G>C, IL-8 +396 G>T, and TGF-β1 +869 G>C and +915 G>C polymorphisms were associated with susceptibility to HPV infection in women from north-east (Pernambuco) Brazil. We analysed 108 healthy uninfected women (HC) and 108 HPV-positive women with cervical lesions. Genetic polymorphisms were assessed using Sanger sequencing and polymerase chain reaction-restriction fragment length polymorphism. Comparison of the distribution of the genotypic and allelic frequencies of the IL-18 +396 T>G polymorphism between HPV infected woman an uninfected controls showed that the GG genotype and G allele were both more frequent in the HC group, and were associated with protection from HPV infection (p = 0.0015; OR = 0.29 CI95% = 0.13-0.61; p = 0.0005; OR = 0.45 CI95% 0.29-0.7, respectively). Individuals from the control group could have previously had HPV infection that was spontaneously eliminated; however, it was undetectable at the time of sample collection. Based on our findings, we hypothesize that the IL-8 +396 G>T polymorphism could interfere with susceptibility to HPV infection, by modulating the ability of immune system to fight the virus.


Subject(s)
Humans , Female , Adolescent , Adult , Middle Aged , Aged , Young Adult , Cervical Intraepithelial Neoplasia/genetics , Interleukin-6/genetics , Interleukin-8/genetics , Papillomavirus Infections/genetics , Polymorphism, Single Nucleotide , Transforming Growth Factor beta1/genetics , Uterine Cervical Neoplasms/genetics , Alleles , Base Sequence , Brazil , Cervical Intraepithelial Neoplasia/virology , Cross-Sectional Studies , DNA, Viral/analysis , Gene Frequency , Genetic Predisposition to Disease , Papillomavirus Infections/virology , Polymerase Chain Reaction , Uterine Cervical Neoplasms/virology
8.
Rev. bras. reumatol ; 56(5): 414-420, Sept.-Oct. 2016. tab, graf
Article in English | LILACS | ID: lil-798103

ABSTRACT

ABSTRACT Background: Rheumatoid arthritis is a widely prevalent autoimmune disorder with suggested genetic predisposition. Objectives: The aim of this study is to detect the pattern of genetic polymorphism of methylene tetrahydrofolate reductase (MTHFR C677 T and A1298 C), transforming growth factor-β1 (TGF-β1 T869 C) and lymphotoxin-α (LT-α A252G) in patients having rheumatoid arthritis and correlate these patterns to disease activity and serum levels of tumor necrosis factor-alpha (TNF-α), B-Cell Activating Factor (BAFF), and osteopontin. Methods: A total of 194 subjects, 90 controls and 104 patients with rheumatoid arthritis were genotyped for MTHFR C677 T and A1298 C, TGF-β1 T869 C and LT-α A252G polymorphisms using a methodology based on PCR-RFLP. Also serum levels of TNF-α, osteopontin and BAFF were measured by ELISA kits. Results: The CT genotype and T allele of MTHFR C677 T and GG genotype and G allele of LT-α A252G are associated with the risk of RA and with higher levels of the pro-inflammatory cytokine, TNF-α in patients with rheumatoid arthritis. Conclusion: Our findings suggest that there is association between MTHFR C677 T and LT-α A252G genes polymorphisms and increased risk of RA in this sample of Egyptian population.


RESUMO Antecedentes: A artrite reumatoide é uma doença autoimune amplamente prevalente com sugerida predisposição genética. Objetivos: Detectar o padrão de polimorfismo dos genes metilenotetrahidrofolato redutase (MTHFR C677 T e A1298 C), fator de crescimento transformador β1 (TGF-β1 T869 C) e linfotoxina-α (LT-α A252G) em pacientes com artrite reumatoide e correlacionar esses padrões com a atividade da doença e os níveis séricos de fator de necrose tumoral alfa (TNF-α), fator ativador de linfócitos B (BAFF) e osteopontina. Métodos: Foram genotipados 194 indivíduos – 90 controles e 104 com artrite reumatoide – à procura de polimorfismos dos genes MTHFR C677 T e A1298 C, TGF-β1 T869 C e LT-α A252G com uma metodologia baseada na PCR-RFLP. Mensuraram-se também os níveis séricos de TNF-α, osteopontina e BAFF com kits de Elisa. Resultados: O genótipo CT e o alelo T do MTHFR C677 T e o genótipo GG e alelo G do LT-α A252G estão associados ao risco de AR e a níveis mais elevados da citocina pró-inflamatória TNF-α em pacientes com artrite reumatoide. Conclusão Os achados do presente estudo sugerem que há associação entre os polimorfismos dos genes MTHFR C677 T e LT-α A252G e um risco aumentado de AR nessa amostra da população egípcia.


Subject(s)
Humans , Arthritis, Rheumatoid/genetics , Lymphotoxin-alpha/genetics , Methylenetetrahydrofolate Reductase (NADPH2)/genetics , Transforming Growth Factor beta1/genetics , Arthritis, Rheumatoid/epidemiology , Transforming Growth Factors , Genetic Predisposition to Disease/genetics , Polymorphism, Single Nucleotide/genetics , Egypt
9.
Clinics ; 71(6): 325-331, tab, graf
Article in English | LILACS | ID: lil-787427

ABSTRACT

OBJECTIVES: We evaluated mRNA expression levels of genes that encode TGF-β1; the TGF-β1 receptor; the collagen-modifying enzymes LOX, PLOD1, and PLOD2; and the extracellular matrix proteins COMP, FN1, TNC and TNXB in synovial/capsule specimens from patients with idiopathic adhesive capsulitis. Possible associations between the measured mRNA levels and clinical parameters were also investigated. METHODS: We obtained glenohumeral joint synovium/capsule specimens from 9 patients with idiopathic adhesive capsulitis who had not shown improvement in symptoms after 5 months of physiotherapy. Adhesive capsulitis was confirmed in all patients by magnetic resonance imaging. We also obtained specimens from 8 control patients who had underwent surgery for acute acromioclavicular joint dislocation and who had radiological indication of glenohumeral capsule alteration based on arthroscopic evaluation. mRNA expression in the synovium/capsule specimens was analyzed by quantitative reverse transcription PCR. The B2M and HPRT1 genes were used as references to normalize target gene expression in the shoulder tissue samples. RESULTS: The synovium/capsule samples from the patients with adhesive capsulitis had significantly higher TNC and FN1 expression than those from the controls. Additionally, symptom duration directly correlated with expression of TGFβ1 receptor I. CONCLUSION: Elevated levels of TNC and FN1 expression may be a marker of capsule injury. Upregulation of TGFβ1 receptor I seems to be dependent on symptom duration; therefore, TGFβ signaling may be involved in adhesive capsulitis. As such, TNC, FN1 and TGFβ1 receptor I may also play roles in adhesive capsulitis by contributing to capsule inflammation and fibrosis.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Young Adult , Bursitis/metabolism , Fibronectins/metabolism , Shoulder Joint/metabolism , Synovial Membrane/metabolism , Tenascin/metabolism , Transforming Growth Factor beta1/genetics , Acromioclavicular Joint/injuries , Acromioclavicular Joint/metabolism , Bursitis/genetics , Case-Control Studies , Extracellular Matrix Proteins/metabolism , Gene Expression , Joint Dislocations/metabolism , Pilot Projects , RNA, Messenger/metabolism , Transforming Growth Factor beta1/metabolism
10.
Article in English | WPRIM | ID: wpr-36805

ABSTRACT

BACKGROUND: Mutations in the transforming growth factor beta-induced gene (TGFBI) are major causes of genetic corneal dystrophies (CDs), which can be grouped into TGFBI CDs. Although a few studies have reported the clinical and genetic features of Korean patients with TGFBI CD, no data are available regarding the frequency and spectrum of TGFBI mutations in a consecutive series of Korean patients with clinically diagnosed CDs. METHODS: Patients with any type of CD, who underwent both ophthalmologic examination and TGFBI gene analysis by Sanger sequencing at a tertiary care hospital in Seoul, Korea from 2006 to 2013, were enrolled in this study. RESULTS: Among a total of 89 patients, 77 (86.5%) were diagnosed as having clinical TGFBI CD. Seventy-three out of 74 patients (98.6%) with granular CD type 2 (GCD2), had the p.R124H mutation. Of particular note, one patient with rapidly progressive CD had the p.R124H mutation as well as a novel nonsense variant with unknown clinical significance (p.A179*). In three patients with lattice CD type 1 (LCD1), one known mutation (p.R124C) and two novel variants (p.L569Q and p.T621P) in the TGFBI gene were identified. CONCLUSIONS: This study provides epidemiological insight into CDs in a Korean population and reaffirms that GCD2 is the most common TGFBI CD phenotype and that p.R124H is the only mutation identified in patients with GCD2. In addition, we broaden the spectrum of TGFBI mutations by identifying two novel missense variants in patients with LCD1.


Subject(s)
Adolescent , Adult , Aged , Asian Continental Ancestry Group/genetics , Corneal Dystrophies, Hereditary/diagnosis , DNA Mutational Analysis , Female , Humans , Male , Middle Aged , Phenotype , Polymorphism, Single Nucleotide , Republic of Korea , Retrospective Studies , Transforming Growth Factor beta1/genetics , Young Adult
11.
Braz. j. med. biol. res ; 47(11): 966-971, 11/2014. tab, graf
Article in English | LILACS | ID: lil-723904

ABSTRACT

Erythropoietin (EPO) has been well characterized as a renal glycoprotein hormone regulating red blood cell production by inhibiting apoptosis of erythrocyte progenitors in hematopoietic tissues. EPO exerts regulatory effects in cardiac and skeletal muscles. Duchenne muscular dystrophy is a lethal degenerative disorder of skeletal and cardiac muscle. In this study, we tested the possible therapeutic beneficial effect of recombinant EPO (rhEPO) in dystrophic muscles in mdx mice. Total strength was measured using a force transducer coupled to a computer. Gene expression for myostatin, transforming growth factor-β1 (TGF-β1), and tumor necrosis factor-α (TNF-α) was determined by quantitative real time polymerase chain reaction. Myostatin expression was significantly decreased in quadriceps from mdx mice treated with rhEPO (rhEPO=0.60±0.11, control=1.07±0.11). On the other hand, rhEPO had no significant effect on the expression of TGF-β1 (rhEPO=0.95±0.14, control=1.05±0.16) and TNF-α (rhEPO=0.73±0.20, control=1.01±0.09). These results may help to clarify some of the direct actions of EPO on skeletal muscle.


Subject(s)
Animals , Male , Down-Regulation/drug effects , Erythropoietin/therapeutic use , Gene Expression/drug effects , Muscular Dystrophy, Duchenne/drug therapy , Myostatin/metabolism , Recombinant Proteins/therapeutic use , Disease Models, Animal , Dystrophin/deficiency , Mice, Inbred mdx , Muscle Strength/drug effects , Muscle, Skeletal/metabolism , Muscular Dystrophy, Duchenne/metabolism , Myostatin/genetics , Phenotype , Real-Time Polymerase Chain Reaction , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/metabolism , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism
12.
Braz. j. med. biol. res ; 47(10): 850-857, 10/2014. tab, graf
Article in English | LILACS | ID: lil-722172

ABSTRACT

We previously described a selective bile duct ligation model to elucidate the process of hepatic fibrogenesis in children with biliary atresia or intrahepatic biliary stenosis. Using this model, we identified changes in the expression of alpha smooth muscle actin (α-SMA) both in the obstructed parenchyma and in the hepatic parenchyma adjacent to the obstruction. However, the expression profiles of desmin and TGF-β1, molecules known to be involved in hepatic fibrogenesis, were unchanged when analyzed by semiquantitative polymerase chain reaction (RT-PCR). Thus, the molecular mechanisms involved in the modulation of liver fibrosis in this experimental model are not fully understood. This study aimed to evaluate the molecular changes in an experimental model of selective bile duct ligation and to compare the gene expression changes observed in RT-PCR and in real-time quantitative PCR (qRT‐PCR). Twenty-eight Wistar rats of both sexes and weaning age (21-23 days old) were used. The rats were separated into groups that were assessed 7 or 60 days after selective biliary duct ligation. The expression of desmin, α-SMA and TGF-β1 was examined in tissue from hepatic parenchyma with biliary obstruction (BO) and in hepatic parenchyma without biliary obstruction (WBO), using RT-PCR and qRT‐PCR. The results obtained in this study using these two methods were significantly different. The BO parenchyma had a more severe fibrogenic reaction, with increased α-SMA and TGF-β1 expression after 7 days. The WBO parenchyma presented a later, fibrotic response, with increased desmin expression 7 days after surgery and increased α-SMA 60 days after surgery. The qRT‐PCR technique was more sensitive to expression changes than the semiquantitative method.


Subject(s)
Animals , Female , Male , Actins/metabolism , Cholestasis/complications , Desmin/metabolism , Liver Cirrhosis/etiology , Liver/metabolism , Real-Time Polymerase Chain Reaction/methods , Transforming Growth Factor beta1/metabolism , Analysis of Variance , Actins/genetics , Biliary Atresia , Bile Ducts/surgery , Collagen Type I/biosynthesis , Disease Models, Animal , Desmin/genetics , Gene Expression , Ligation , Liver Cirrhosis/metabolism , Liver/surgery , Rats, Wistar , Transforming Growth Factor beta1/genetics
13.
Clinics ; 68(7): 1004-1009, jul. 2013. tab, graf
Article in English | LILACS | ID: lil-680710

ABSTRACT

OBJECTIVES: The aim of this cross-sectional study was to evaluate whether interleukin 10 (IL10) and transforming growth factor β1 (TGFβ1) gene polymorphisms were associated with persistent IgE-mediated cow's milk allergy in 50 Brazilian children. The diagnostic criteria were anaphylaxis triggered by cow's milk or a positive double-blind, placebo-controlled food challenge. Tolerance was defined as the absence of a clinical response to a double-blind, placebo-controlled food challenge or cow's milk exposure. METHOD: The genomic DNA of the 50 patients and 224 healthy controls (HCs) was used to investigate five IL10 gene polymorphisms (-3575A/T, -2849A/G, -2763A/C, -1082G/A, -592C/A) and one TGFβ1 polymorphism (-509C/T). RESULTS: Among the five IL10 polymorphisms analyzed, homozygosis for the G allele at the -1082 position was significantly higher in the patients compared with the healthy controls (p = 0.027) and in the persistent cow's milk allergy group compared with the healthy controls (p = 0.001). CONCLUSIONS: Homozygosis for the G allele at the IL10 -1082G/A polymorphism is associated with the persistent form of cow's milk allergy. .


Subject(s)
Child , Female , Humans , Male , Immunoglobulin E/immunology , /genetics , Milk Hypersensitivity/genetics , Polymorphism, Genetic/genetics , Polymorphism, Single Nucleotide/genetics , Transforming Growth Factor beta1/genetics , Brazil , Case-Control Studies , Cross-Sectional Studies , Gene Frequency , Logistic Models , Milk Hypersensitivity/immunology , Polymerase Chain Reaction , Risk Factors
14.
Rev. AMRIGS ; 56(3): 204-212, jul.-set. 2012. tab
Article in Portuguese | LILACS | ID: biblio-848047

ABSTRACT

Introdução: O fator de crescimento transformante beta-1 (TGFß1) é uma citocina multifuncional que regula a proliferação, a diferenciação e a formação da matriz extracelular de vários tipos de células. Existem evidências de que os polimorfismos -509C>T e 869T>C no gene do TGFß1 alteram a sua expressão gênica e podem estar envolvidos na patogênese das complicações crônicas do diabetes mellitus tipo 2 (DM2). O objetivo deste estudo foi analisar o papel dos polimorfismos funcionais -509C>T e 869T>C no gene do TGFß1 na patogênese da retinopatia (RD) e da nefropatia (ND) diabéticas, em pacientes com DM2. Métodos: Foi realizado um estudo de caso-controle aninhado a um estudo transversal. Foram selecionados pacientes caucasoides com DM2 que realizaram exames clínicos, laboratoriais e uma entrevista com questionário padronizado. A genotipagem dos polimorfismos foi realizada por meio de PCR. Resultados: As frequências genotípicas e alélicas obtidas para o polimorfismo -509C>T nos pacientes com RD ou ND não diferiram daquelas observadas nos pacientes sem estas complicações. Mas quando a gravidade das complicações foi analisada, observou-se que o alelo C foi menos frequente entre os pacientes com insuficiência renal crônica não tratada com diálise, assim como a frequência de homozigotos para o alelo C foi maior nos pacientes dialíticos. Conclusão: Após a análise multivariada, o genótipo CC permaneceu como um fator de risco associado com a progressão da ND (RC = 2,68, IC 95% 1,08-6,68). Assim, os resultados sugerem que o polimorfismo 869T>C no gene do TGFß1 está associado com a progressão da ND em pacientes com DM2 (AU)


Introduction: The transforming growth factor-beta 1 (TGFß1) is a multifunctional cytokine that regulates proliferation, differentiation and extracellular matrix formation of multiple cell types. There is evidence that polymorphisms -509C> T and 869T> C in the TGFß1 gene alter its gene expression and may be involved in the pathogenesis of chronic complications of diabetes mellitus type 2 (DM2). The aim of this study was to analyze the role of functional polymorphisms -509C> T and 869T> C in the TGFß1 gene in the pathogenesis of diabetic retinopathy (DR) and diabetic nephropathy (DN) in patients with DM2. Methods: We conducted a case-control study nested in a cross-sectional study. We selected Caucasian patients with DM2 who underwent clinical and laboratory tests and an interview with a standardized questionnaire. Polymorphism genotyping was performed by PCR. Results: The allele and genotype frequencies obtained for polymorphism -509C> T in patients with DR or DN did not differ from those observed in patients without these complications. But when the severity of complications was analyzed, the C allele was found to be less frequent among patients with chronic renal failure untreated with dialysis, and the frequency of homozygous for the C allele was higher in dialysis patients. Conclusion: After multivariate analysis, the CC genotype remained a risk factor associated with progression of DN (OR = 2.68, 95% CI 1.08 to 6.68). Thus, the results suggest that polymorphism 869T> C in THE TGFß1 gene is associated with progression of DN in patients with DM2 (AU)


Subject(s)
Humans , Male , Female , Middle Aged , Aged , Diabetic Nephropathies/genetics , Diabetic Retinopathy/genetics , Genetic Predisposition to Disease/epidemiology , Polymorphism, Genetic/genetics , Transforming Growth Factor beta1/genetics , Case-Control Studies , Cross-Sectional Studies , Diabetes Mellitus, Type 2/complications , Diabetic Nephropathies/etiology , Diabetic Retinopathy/etiology
15.
Braz. j. med. biol. res ; 44(2): 91-99, Feb. 2011. ilus, tab
Article in English | LILACS | ID: lil-573653

ABSTRACT

Searching for effective Smad3 gene-based gene therapies for hepatic fibrosis, we constructed siRNA expression plasmids targeting the rat Smad3 gene and then delivered these plasmids into hepatic stellate cells (HSCs). The effect of siRNAs on the mRNA levels of Smad2, Smad3, Smad4, and collagens I-α1, III-α1 and IV-α1 (Colα1, Col3α1, Col4α1, respectively) was determined by RT-PCR. Eighty adult male Sprague-Dawley rats were randomly divided into three groups. Twice a week for 8 weeks, the untreated hepatic fibrosis model (N = 30) and the treated group (N = 20) were injected subcutaneously with 40 percent (v/v) carbon tetrachloride (CCl4)-olive oil (3 mL/kg), and the normal control group (N = 30) was injected with olive oil (3 mL/kg). In the 4th week, the treated rats were injected subcutaneously with liposome-encapsulated plasmids (150 µg/kg) into the right liver lobe under general anesthesia once every 2 weeks, and the untreated rats were injected with the same volume of buffer. At the end of the 6th and 8th weeks, liver tissue and sera were collected. Pathological changes were assessed by a semi-quantitative scoring system (SSS), and a radioimmunoassay was used to establish a serum liver fibrosis index (type III procollagen, type IV collagen, laminin, and hyaluronic acid). The mRNA expression levels of the above cited genes were reduced in the HSCs transfected with the siRNA expression plasmids. Moreover, in the treated group, fibrosis evaluated by the SSS was significantly reduced (P < 0.05) and the serum indices were greatly improved (P < 0.01). These results suggest that Smad3 siRNA expression plasmids have an anti-fibrotic effect.


Subject(s)
Animals , Male , Rats , Down-Regulation/genetics , Hepatic Stellate Cells/metabolism , Liver Cirrhosis, Experimental/metabolism , RNA, Messenger/genetics , RNA, Small Interfering/therapeutic use , /metabolism , Carbon Tetrachloride , Collagen/metabolism , Liposomes , Liver Cirrhosis, Experimental/chemically induced , Liver Cirrhosis, Experimental/genetics , Liver Cirrhosis, Experimental/pathology , Plasmids , Radioimmunoassay , Random Allocation , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , RNA Interference , RNA, Messenger/metabolism , Severity of Illness Index , /genetics , Transfection , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/metabolism
16.
Article in English | WPRIM | ID: wpr-102684

ABSTRACT

Granular corneal dystrophy, type II (CGD2; Avellino corneal dystrophy) is the most common corneal dystrophy among Koreans, but its pathophysiology is still poorly understood. Many reports showed that even though the causative mutation is the same TGFBI R124H mutation, there are severe and mild phenotypes of the corneal dystrophy. We also observed the phenotype differences in our samples. For this reason, we focused our effort on the identification of unknown genetic factor related to phenotype variation. A total 551 individuals from 59 families were genotyped with SNP chip and used in genome-wide linkage analysis. From single-point linkage analyses, we confirmed the known 5q31 region for TGFBI gene, and selected novel nine candidate loci for CGD2. In simulation analysis, the only 3q26.3 region including neuroligin 1 gene (NLGN1) was supported by empirical statistic significance. To investigate the effect of genetic heterogeneity in linkage analysis, we classified CGD2 families into two subgroups. Although we could not find a significant evidence for correlation between the 3q26.3 region and CGD2 phenotypes, this first genome-wide analysis with CGD2 families in Korea has a very important value for offering insights in genetics of CGD2. In addition, the co-segregating loci with CGD2 including 3q26.3 would be a good target for further study to understand the pathophysiology of CGD2.


Subject(s)
Cell Adhesion Molecules, Neuronal/genetics , Chromosomes, Human, Pair 3/genetics , Chromosomes, Human, Pair 5/genetics , Computer Simulation , Corneal Dystrophies, Hereditary/genetics , Female , Genetic Linkage , Genetic Loci , Genome-Wide Association Study , Genotype , Humans , Male , Models, Genetic , Polymorphism, Single Nucleotide , Transforming Growth Factor beta1/genetics
17.
Article in English | WPRIM | ID: wpr-169276

ABSTRACT

BACKGROUND/AIMS: Transforming growth factor beta1 (TGF-beta1) is a key cytokine in the production of extracellular matrix. A genetic polymorphism at codon 10 of the TGF-beta1 gene is associated with liver fibrosis. We investigated the effect of genetic polymorphisms at codon 10 on the development of alcoholic liver cirrhosis (ALC). METHODS: In total, 119 controls and 182 patients with ALC, were enrolled in the study. Clinical and laboratory data including total lifetime alcohol intake were collected at enrollment. The genotype at codon 10 was determined for each patient by single-strand conformation polymorphism. RESULTS: There were three types of genetic polymorphism at codon 10: homozygous proline (P/P), heterozygous proline/leucine (P/L), and homozygous leucine (L/L). Among the controls, the proportions of P/P, P/L, and L/L were 26.1%, 44.5%, and 29.4%, respectively in the ALC group, these proportions were 23.1%, 43.4%, and 33.5%, respectively. The genotype distribution did not differ between the controls and the ALC group. In the ALC group, age, total lifetime alcohol intake, and distribution of Child-Pugh class did not differ with the genotype. Of the male patients with ALC (n=164), the proportions of P/P, P/L, and L/L were 20.1%, 44.5%, and 35.4%, respectively the genotype distribution did not differ between the male controls and the male ALC patients. CONCLUSIONS: The genotype at codon 10 in TGF-beta1 does not appear to influence the development of ALC. Further study is needed to investigate other genetic factors that influence the development of ALC in patients with chronic alcohol intake.


Subject(s)
Aged , Alcohol Drinking , Codon , Female , Genotype , Heterozygote , Homozygote , Humans , Liver Cirrhosis, Alcoholic/genetics , Male , Middle Aged , Polymorphism, Genetic , Transforming Growth Factor beta1/genetics
18.
Clinics ; 66(7): 1143-1147, 2011. ilus, tab
Article in English | LILACS | ID: lil-596899

ABSTRACT

OBJECTIVE: To evaluate the correlation between transforming growth factor beta (TGF-β1) expression and prognosis in prostate cancer. PATIENTS AND METHODS: TGF-β1 expression levels were analyzed using the quantitative real-time polymerase chain reaction to amplify RNA that had been isolated from fresh-frozen malignant and benign tissue specimens collected from 89 patients who had clinically localized prostate cancer and had been treated with radical prostatectomy. The control group consisted of li patients with benign prostate hyperplasia. The expression levels of TGF-β1 were compared between the groups in terms of Gleason scores, pathological staging, and prostate-specific antigen serum levels. RESULTS: In the majority of the tumor samples, TGF-β1 was underexpressed 67.0 percent of PCa patients. The same expression pattern was identified in benign tissues of patients with prostate cancer. Although most cases exhibited underexpression of TGF-β1, a higher expression level was found in patients with Gleason scores >7 when compared to patients with Gleason scores <7(p = 0.002). Among the 26 cases of TGF-β1 overexpression, 92.3 percent had poor prognostic features. CONCLUSIONS: TGF-β1 was underexpressed in prostate cancers; however, higher expression was observed in tumors with higher Gleason scores, which suggests that TGF-β1 expression may be a useful prognostic marker for prostate cancer. Further studies of clinical specimens are needed to clarify the role of TGF-β1 in prostate carcinogenesis.


Subject(s)
Adult , Aged , Humans , Male , Middle Aged , Prostatic Neoplasms/metabolism , Transforming Growth Factor beta1/metabolism , Biomarkers, Tumor/metabolism , Carcinogens/metabolism , Gene Expression , Neoplasm Grading , Prognosis , Prostatectomy , Prostate-Specific Antigen/blood , Prostatic Neoplasms/pathology , Prostatic Neoplasms/surgery , Real-Time Polymerase Chain Reaction , Statistics, Nonparametric , Transforming Growth Factor beta1/genetics
19.
Article in English | IMSEAR | ID: sea-139892

ABSTRACT

Background : Oral submucous fibrosis (OSF) may be considered a collagen metabolic disorder resulting from areca-nut alkaloid exposure and individual variation in collagen metabolism. Due to the complexity of OSF pathogenesis, it is important to elucidate independent and interactive effects of polymorphisms of collagen-related genes on OSF risk. Materials and Methods : This study is focused on seven polymorphisms (SNPs) of transforming growth factor-beta-1 (TGF-beta-1) gene in patients with oral submucous fibrosis (OSF), belonging to south Indian ethnic extraction. The mean age at presentation was 43.9 years, range 23-72 years (n=50, M:F ratio, 2.6:1). DNA samples from 50 subjects of the same ethnic group and comparable demographic features who have had practiced the habit of areca-chewing of almost equal duration, but remained free of disease constituted the controls. All DNA samples were collected progressively and purified from peripheral blood employing standard protocols and tested for SNPs. They included two polymorphisms in the promoter region (C-509T and G-800A), three polymorphisms in exon-1 (Arg25Pro(G915C), Leu10Pro(T869C), Glu47Gly(A979G) and two in 5 ͲUTR regions (C→T(rs13306708) and G→A (rs9282871). The extracted DNA samples along with the primers underwent PCR amplification and the genotypic and allelic frequencies were calculated. All calculations were performed using the SPSS software. The PCR products were purified and subsequently sequenced using Flour S™ multi-imager system (Biorad). The sequenced data were analyzed using the BioEdit sequence analysis software. Results : Out of the seven polymorphisms analyzed, six such as two in the promoter region, three in exon-1 and one in 5¢UTR were found to have a " P" value above 0.05 and hence were not significant. The C→T transition (rs13306708) in the 5¢UTR region recorded a " P" value of 0.03 on comparison and hence was found to be significant. The allelic frequencies for this C→T transition in patients were 68.7% C and 31.2% T (27CC, 15CT, 8TT) and that in controls were 89.5% C and 10.4% T (42CC, 6CT, 2TT). Conclusions : The polymorphism in 5¢UTR C-T in TGF beta 1 gene has a significant association with OSF, being a prime determinant in the pro-angiogenic pathway which has got direct bearing with the pathophysiology of the disease. The proximity of this polymorphism to the transcription site and the associated risk involved is discussed.


Subject(s)
5' Untranslated Regions/genetics , Adenine , Adult , Aged , Areca , Arginine/genetics , Chromosome Mapping , Cytosine , Ethnic Groups/genetics , Exons/genetics , Female , Gene Frequency/genetics , Genotype , Glutamine/genetics , Glycine/genetics , Guanine , Humans , India , Leucine/genetics , Male , Middle Aged , Oral Submucous Fibrosis/genetics , Oral Submucous Fibrosis/immunology , Polymorphism, Single Nucleotide/genetics , Proline/genetics , Promoter Regions, Genetic/genetics , Thymine , Time Factors , Transforming Growth Factor beta1/genetics , Young Adult
20.
Indian J Ophthalmol ; 2010 Jul; 58(4): 328-329
Article in English | IMSEAR | ID: sea-136082

ABSTRACT

An eight-year-old girl, an offspring of a consanguineous marriage presented with multiple anterior stromal geographic corneal opacities in both eyes. She was diagnosed to have superficial variant of granular dystrophy based on the family history, clinical features and mutation of TGF B1 gene. She was treated by alcohol-assisted removal of epithelium followed by mechanical debridement of abnormal deposits. Postoperatively, the cornea in both eyes was clear with no trace of opacity and the patient had an unaided visual acuity of 20/20 partial.


Subject(s)
Amino Acid Substitution , Anti-Bacterial Agents/therapeutic use , Child , Corneal Dystrophies, Hereditary/drug therapy , Corneal Dystrophies, Hereditary/genetics , Corneal Dystrophies, Hereditary/surgery , Debridement/methods , Female , Genetic Variation , Homozygote , Humans , Male , Mutation , Ofloxacin/therapeutic use , Siblings , Transforming Growth Factor beta1/genetics , Treatment Outcome , Visual Acuity
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