Kaempferol promotes wound-healing in diabetic rats through antibacterial and antioxidant effects, devoid of proliferative action

The investigation of novel phytochemicals for the prevention and treatment of infections caused by multidrug-resistant pathogens is gaining attention. The current study evaluated the in-vitro antimicrobial activity of kaempferol against methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. Its in-vivo efficacy in inhibiting these pathogens was determined using an excision wound model in nicotinamide-streptozocin- -induced diabetic rats. Kaempferol displayed an inhibitory effect against the tested bacteria both in vitro and in vivo. It also healed excision wounds at a 1% (w/w) concentration. An increase in antioxidant enzymes in wounded tissue was observed on kaempferol treatment. A reduction in the MRSA and P. aeruginosa counts in wounded tissue together with a reduced epithelization period was observed when compared to the infected control. A thicker epithelium, new capillaries, and a decrease in inflammatory cells were detected by hematoxylin and eosin staining. Furthermore, an increase in collagen fibers and their deposition was observed by Masson's trichrome staining. Kaempferol at 40 µM did not display any toxicity for human keratinocytes grown in media containing high glucose and it did not affect the expression of the pro-healing cytokines genes vascular endothelial growth factor (VEGF) and transforming growth factor-b-1 (TGFß1). Kaempferol displayed antibacterial and antioxidant actions but did not increase the expression of proliferative genes.

Interplay between TGF-b1 and miRNA-122 biomarkers in hepatocellular carcinoma progression in patients with chronic hepatitis C

@#Hepatocellular carcinoma (HCC) is a highly lethal malignancy and clinically validated medications have not yet been developed since there are no reliable diagnostic and prognostic biomarkers. Based on bioinformatics tools, TGF-b1 gene was the first target gene of miRNA-122, therefore this study was intended to assess the potential interconnection between TGF-b1 and miRNA-122 as a diagnostic and prognostic biomarker in the progression of HCC in patients with chronic hepatitis C (CHC) genotype (4). In this study, 100 people were included and split into two groups; group I: CHC patients without HCC that were classified into patients CHC without cirrhosis and CHC cirrhotic patients, group II: CHC patients with HCC, and healthy volunteers as control. The expression of miRNA-122 and TGF-b1 genes were analyzed using Real-Time PCR. An upregulation of miRNA-122 gene in cirrhotic and HCC patients compared to both chronic HCV non-cirrhotic, and cirrhotic patients, while, a decrease in expression of TGF-b1 was found in cirrhotic patients compared to HCV non-cirrhotic patients. Although significantly downregulated in HCC patients. Regression analysis indicated that the expression levels of miRNA-122 and TGF-b1 could be regarded as important indicators of the alterations in cirrhotic and HCC patients versus HCV non-cirrhotic patients, also with the chances of HCC versus cirrhosis patients. Our data indicated an interaction between miRNA-122 and TGF-b1, regulated gene expression and recommended the use of these parameters as noninvasive predictive biomarkers and therapeutic targets for HCV induced liver cirrhosis and HCC.

Involvement of eIF2α in halofuginone-driven inhibition of TGF-β1-induced EMT

Halofuginone (HF) is an extract from the widely used traditional Chinese medicine (TCM) Dichroa febrifugathat facilitates the recovery of wounds and attenuates hepatic fibrosis. However, the role of HF in theepithelial-mesenchymal transition (EMT) of IPEC-J2 cells remains unclear. The current study explored theanti-EMT effect of HF in IPEC-J2 cells and illustrates its molecular mechanism. Transforming growth factorb1 (TGF-b1), as a recognized profibrogenic cytokine, decreased the level of the epithelial marker E-cadherinand increased the level of the mesenchymal markers, such as N-cadherin, fibronectin (FN), vimentin (Vim),and a-smooth muscle actin (a-SMA), in IPEC-J2 cells depending on the exposure time and dose. HF markedlyprevented the EMT induced by TGF-b1. Dissection of the mechanism revealed that HF inhibited IPEC-J2 cellEMT via modulating the phosphorylation of SMAD2/3 and the SMAD2/3-SMAD4 complexnuclear translocation. Furthermore, HF could promote the phosphorylation of eukaryotic translation initiationfactor-2a (eIF2a), which modulates the SMAD signaling pathway. These results suggested that HF inhibitsTGF-b1-induced EMT in IPEC-J2 cells through the eIF2a/SMAD signaling pathway. Our findings suggest thatHF can serve as a potential anti-EMT agent in intestinal fibrosis therapy.

Relationship between the Change of IGF 1 and TGF-β1 Level and Renal Damage in Children with Allergic Purpura / 现代检验医学杂志

Objective To investigate the effects of insulin-like growth factor 1 (IGF 1) and transforming growth factor beta 1 (TGF-β1) in he pathogenesis of children with allergic purpura kidney damage.Methods 135 henoch-schonlein purpura (HSP) children with kidney damage were divided into HSP and HSPN group according to whether associated with renal damage,blood IGF 1,TGF-β1,urinary inhibition C (Cys C),creatinine (SCr) and content of urea nitrogen (BUN) were compared,and blood IGF-1,TGF-β1 and Cys C content of HSPN patients in different pathological grading were compared,the correlation of blood IGF 1,TGF-β1 and Cys C content of the HSPN group were analyzed,Results Blood IGF 1,TGF-β1 and Cys C content of the control group(117.2±18.8 ng/L,164.2±18.4 ng/L,0.9±0.2 mg/L),the HSP group(131.7±19.6 ng/L,282.1±28.3 ng/L,1.1±0.2 mg/L) and the HSPN group (205.3±24.5 ng/L,489.2±32.7 ng/L,1.3±0.3 mg/L) showed a trend of increasing gradually (F=4.824～45.066,P value＜0.01),the HSP group and the HSPN group were higher than that of the control group (q=3.397～58.931,P value＜0.01),the HSPN group was higher than that of the HSP group (q=16.997,35.193,P value＜0.01),the difference was statistically significant.Blood IGF-1 (level Ⅱ 175.6 ± 20.4 ng/L,level m198.5±23.3 ng/L,level Ⅳ241.7±25.1 ng/L),TGF-β1(level Ⅱ 392.8±38.9 ng/L,level Ⅲ 481.3± 44.03 ng/L,level Ⅳ 537.6±42.9 ng/L),Cys C (level 11 1.1±0.3 mg/L,level Ⅲ 1.3±0.4 mg/L,level Ⅳ1.6±0.4 mg/L) content of children with HSPN increased with the increase of renal pathology classification (F=6.594～ 28.317,P value ＜0.01),blood IGF-1,TGF-β1 and Cys C content of kidney pathology classification of Ⅵ level in children was higher than that of the level of Ⅱ and Ⅲ in children (q=2.415～11.818,P＜0.05 or P＜0.01),while the contern of blood IGF-1,TGF-β1 and Cys C of level Ⅲ in children was higher than that of the level Ⅱ in children (q=2.577～6.244,P＜0.05 orP＜ 0.01),the difference was statistically significant.Blood IGF-1,TGF-β1 content of children with HSPN were positively correlated with Cys C content of children (r=0.648,0.719,P＜0.05),but blood IGF 1 content was significantly positive correlated with TGF-β1 content (r=0.748,P＜0.05).Conclusion IGF 1 and TGF-β31 partieipated in the pathogenesis of HSPN,and both were correlated with the degree of the pathological damage.

Results of the bone healing performance of Osteo-Calcium 5 preparation tested by internal fixation surgery patients / Монголын Анагаах Ухаан

INTRODUCTION: Nowadays, the population of noncommunicable sickness is growing rapidly changing lifestyle is one example of this is the widening of accidents-related bone fractures.The National Center for Trauma Treatment of inpatient hospital from a total fractures 10260 or 30.8%, of which 6777 or 66.05% of the marrow bone fractures as well as drugs used in refractive rarely occupied. PURPOSE: To consider a result of the use of Osteocalcium 5 medical preparation for patients who were diagnosed diaphyseal fracture of the tibia and treated by Intramedullar Osteosyntesis (internal fixation)OBJECTIVES:1. To establish process of fractured bone remodeling to original bone contour by TGF-b1 cytokine dynamics in serum2. To establish process of fractured bone remodeling to original bone contour by alkaline phosphatase dynamics in serumMATERIALS AND METHODS: Within the framework of the survey 28 patients of the “Bone, joint and limb section” Section of the National Institute of Traumatology, aged between 25-60 years, were involved in the present survey. Survey participants were classified in 2 groups according to “Krischner wire” position and their Xray’s images. An accordance to the methodology of James Balch and Mark Stengller (2011) daily dosage of the Osteocalcium 5 preparation for test and comparing patients was determined as following introduced: for test group-twice a day before meal 1.5 grams, for comparing group–Calcium D3Nycomed preparation three times a day during 60 days. Transforming growth factor-beta 1(TGFb1) and Alkaline phosphatase was equal of 3, 14, 42 fraction days.RESULT: The present diagram shows that according to the curve TGF-b1 the bone excrescence performance of the Osteocalcium 5 preparation (0.0001) is statically increasing at 14 days and after 42 days it is statically decreasing and the calculated discrepancy (p<0.015) reveals and proofs the impact of its bone excrescence. Alkaline phosphatase in serum 14 days after the fracture Osteocalcium 5 pills used in chapter Nycomed Calcium D3 drank chapters (116,06±35U/L) compared to a statistical difference (p<0.044).CONCLUSION: Preparations Ostyeokalitsi 5 osteoblast chondrocyte cell activity and enable them synthesized by the cells in the TGF-B1 osteoblast line Alkaline phosphatase synthesized in increased serum concentrations of the bone heal affect signaling molecules involved are determined that quickens recovery.

Avaliação de sete protocolos para obtenção de plasma rico em plaquetas na espécie equina / Evaluation of seven platelet-rich plasma processing protocols in the equine species

O presente estudo teve por objetivo avaliar a capacidade de concentração plaquetária e sua correlação com os níveis do fator de crescimento TGF-B1, a presença de leucócitos e de hemácias nos diferentes protocolos utilizados na obtenção do plasma rico em plaquetas (PRP) de equinos, através do método manual. Dez equinos, sadios, com idade média de 7 anos (±2,39), pesando em média 500kg (±67,1) foram utilizados neste estudo. Os protocolos testados variaram na velocidade e no tempo nas duas centrifugações. As variáveis analisadas nas amostras de PRP foram: concentração de plaquetas, presença de leucócitos e hemácias, e níveis de TGF-β1 quantificados pelo teste ELISA. Os protocolos testados não diferiram na capacidade de concentração de plaquetas e nos níveis de TGF-β1. Entretanto, houve diferença significava entre o protocolo I e os demais por este apresentar maior número de hemácias e leucócitos nas amostras de PRP, sendo por esse motivo considerado um protocolo inadequado para processamento do volume de sangue utilizado. Os demais protocolos podem ser utilizados para obtenção de PRP terapêutico em equinos.

PRP is plasma that contains a high numbers of platelets and growth factors in a small volume. The aim of this study was to evaluate seven different protocols to obtain PRP by the manual method according to their capacity to concentrate platelets, leukocyte and erythrocyte contamination and correlation between platelet count and TGF-β1 growth factor levels in PRP samples. Ten healthy horses with a mean age of 7 years (±2.39), weighing on average 500 kg (±67.1) were used in this study. The protocols tested varied according to the speed and time used at the two centrifugations. PRP samples were analyzed regarding platelet concentration, leukocyte and erythrocyte contamination and TGF-β1 levels quantified by ELISA. No significant differences among protocols were observed regarding the ability to concentrate platelets and TGF-β1. However, protocol I showed significantly higher erythrocyte and leukocyte counts in PRP samples than the other protocols, reason why it was considered inadequate for the volume of blood processed in this experiment. The remaining protocols are suitable for extracting PRP.

Alcohol epitheliectomy with mechanical debridement in a case of granular corneal dystrophy with r555w homozygous mutation of TGF B1 gene.

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.

Transforming growth factor-β-1 polymorphisms are infrequent but exist at selected loci in oral submucous fibrosis.

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.

Double Plate Occipitocervical Fusion After Failed Posterior Fusion C 1-2 With Wiring / 대한척추외과학회지

A 31-year-old female complained of neck pain and limitation in neck motion. She had a 3 month history of treatment with Halovest at another hospital for a fracture of the odontoid process due to a car accident. The patient complained of persistent pain and limitation in neck motion following the cessation of Halovest. A dynamic radiograph demonstrated instability on C1-2 and she underwent a posterior cervical fusion with wiring. A wound infection developed, and loosening of the wire and lysis of the posterior arch at C1-2 were seen on a follow up plain radiograph 2 months postoperatively. She was transferred to our hospital where she underwent occipitocervical fusion with a double plate after control of the infection. There were rigid fixations of the plate and bone union on a follow up radiograph 24 months postoperatively.

Mesenchymanl Stem Cell Based Intradiscal Gene Therapy: Therapeutic Implication in Degenerative Disc Disease / 대한척추외과학회지

STUDY DESIGN: In-vitro experiments using human mesenchymal stem cells (MSCs), intervertebral disc (IVD) cells and type 5 adenovirus/transforming growth factor-beta1 construct (Ad/TGF-beta1). OBJECTIVES: To determine the effect of MSC-based gene therapy for matrix regeneration of IVD cells. SUMMARY OF LITERATURE REVIEW: MSCs are known to be multipotent in tissue regeneration. In degeneration of IVD, cellular replacement with genetic modification other than that of IVD cells may prove an enhanced mechanism for the regeneration of MATERIALS AND METHODS: MSCs and IVD cells were cultured and an adenovirus construct containing TGF-beta1 cDNA (Ad/TGF-beta1) was also produced. In the first step, the MSCs were transduced with Ad/TGF-beta1, then mixed with IVD cells in various proportions and three dimensionally cultured. [methyl-(3)H]Thymidine and [(35)S]Sulfur incorporation for DNA and proteoglycan synthesis, respectively, were measured. RT-PCR was performed to assess the aggrecan and collagen types I and II mRNA RESULTS: Mixed cultures of MSC and IVD cells showed relatively similar amounts of newly synthesized proteoglycan compared with cultures of IVD cells only. In mixed cultures transduced with Ad/TGF-beta1, there were significant decreases in newly synthesized proteoglycan with increasing the proportions of MSCs, which was also found with the aggrecan and collagen type II mRNA expressions. However, the collagen type I mRNA expression increased with increased proportions of MSCs transduced with Ad/TGF-beta1. CONCLUSION: Cell therapy with MSCs and IVD cells provided a mechanism for cellular augmentation. However, MSC-based gene therapy coupled with IVD cells did not maintain a chondrogenic phenotype.

Change of the Effect of TGF-beta1 on Physeal Chondrocytes According to Culture Methods in Vitro / 대한정형외과학회잡지

PURPOSE: We intended to check the growth rates and phenotypic markers of chondrocytes in the dedifferentiated cells cultivated in various conditions in order to establish the ideal culture system for implantation. MATERIALS AND METHODS: Culturing rabbit chondrocytes from proximal tibia, we checked the phenotypes at first, second, and third week. Then we cultured the chondrocytes in different circumstances such as monolayer or three dimensional gel in the presence or abscence of TGF-B1, and checked the growth rates and phenotypic markers. RESULTS: There was no difference in growth rates and mRNA level of type I, type II collagen and aggrecan between the cells cultured in monolayer and three dimensional gel of collagen. However, the responses of the cells to TGF-B1, were quite different between these two groups. In monolayer culture, the expression of type I collagen was depressed by TGF-B1 while the growth rate was markedly increased. Oppositely in three dimensional culture, the mRNA level of type I collagen was markedly increased and the growth rate was completely suppressed by TGF-B1. The expression of type II collagen could be detected only in TGF-B1-treated cells cultured in three dimensional gel for 4 or more days. The mRNA level of aggrecan was also increased by TGF-B1, in the cells cultured in three dimensional gel. CONCLUSIONS: These results suggest that the number of chondrocytes can be efficiently expanded by culturing the cells in monolayer and the phenotypes of chondrocyte can be restored by culturing the cells in three dimensional gel containing TGF-B1. The application of semi-solid gel containing differentiated chondrocytes in physeal implantation should be further evaluated

Study on Growth Suppression Effect of Vetamin D3 Mediated by Transfrorming Growth Factor-B1(TGF-B1) in Acute Myelogenous Leukemic Cell / 대한암학회지

PURPOSE: Vitamin D3 was shown to arrest the growth of acute myelogenous leukemic cells and transforming growth factor- B1 (TGF- B1) was reported to be involved in the mechanism of vitamin D3. We studied the growth inhibitory effect of 1,25(OH)2-vitamin D3(C) and its analogue (EB1089) in leukemic cell lines and the changes in the secretion or the activation of TGF-B1 in the supernatant and the status of TGF-B1 type II receptor. MATERIALS AND METHODS: Growth inhibition by vitamin D3 and TGF-B1 in 5 leukemic cell lines (HEL, HL-60, U937, KG-1, K562) were assessed with clonogenic and [3H]thymidine assay respectively. TGF-B type II receptor status was examined by Southern and Northern blotting. The concentrations of TGF- B1 in the supernatant were quantitated by enzyme immunoassay. RESULTS: The growth of HEL, HL-60, U937 were inhibited in a dose-dependent fashion by both C and EB1089, more markedly by the latter. Anti-TGF-B neutralizing antibody partially reversed the growth inhibition. TGF-B1 markedly inhibited the growth of HEL, U937, KG-1, SNU-16 dose dependently while HL-60 and K562 showed no growth inhibition. HEL secreted latent TGF- 1 and HL-60 activated latent TGF- B1 or secreted active TGF-B1 irrespective of the treatment with vitamin D3. In U937, vitamin D3 increased the concentration of both active and latent TGF-B1. Deletion or abnormal expression of TGF- B type II receptor gene was not found in the 5 cell lines examined. CONCLUSION: Vitamin D3 has various pattern of growth inhibition in acute myelogenous leukemia and inhibits the growth of some cell lines by secretion or activation of TGF-B1. Abnormality of TGF-B type II receptor DNA or mRNA seems to be rare.

GEnetic Change in Transforming Growth Factor-B (TGF-B) Receptor Type I and Type II Genes with Resistance to TGF-B of Human Breast Cancer Cells / 대한암학회지

PURPOSE: Transforming growth factor-Bs (TGF-Bs) are prototypic multifunctional negative growth factors that inhibit the growth of many cell types. TGF-B type I and II receptors(RI, RII) are transmembrane receptors containing cytoplasmic serine/ threonine kinase domain and have been implicated in mediating TGF-B activity. Because a heteromeric complex of RI and RII is required for TGF-B signal transduction, cancer cells may reduce the expression of either RI or RII to escape from growth inhibition of TGF-B. We examined the correlation between the growth inhibitory activity of TGF-B1 and the genetic expression of RI &RII genes in human breast cancer cell lines. MATERIALS AND METHODS: We examined the growth inhibitory activity of TGF-B1 in 5 breast cancer cell lines by incorporation of [3H] thymidine. To investigate the correlation between TGF-B1 insensitivity and genetic change of TGF-B receptor genes (RI, RII), Southem blot analysis, Northern blot analysis, and Western blot analysis were performed. We also examined whether microsatellite instability(RER) was associated with RII mutation. RESULTS: We found that 3 breast cancer cell lines (MCF-7, YCC-B101, YCC-B151) were resistant to growth inhibitory effect of TGF-B1. MCF-7 cell line expressed no detectable RII mRNA and RII protein, but showed normal structure of RII gene and normal expression of RI gene. And we did not find any abnormal expression of mRNA, protein, and genetic structure of RI &RII in YCC-B101 and YCC-B151. CONCLUSION: Our results suggest that aquired resistance to the growth inhibitory effect of TGF-B1> could be transcription regulation system of RII in MCF-7 cell line, and could be postreceptor signal transduction pathway in YCC-B101 and YCC-B151 cell lines.

Effects of Steroids and vitamin D Derivatives on Differentiation and Proliferation of Cultrured Osteoblast Cells / 대한정형외과학회잡지

This study was designed to evaluate of the effects ofB-estradiol, progesterone, dexamethasone, hydrocortisone, calcitriol and alfacalcidol on the cell proliferation and differentiation in the rat osteoblast-like osteosarcoma cells ROS 17/2.8. The proliferation of ROS 17/2.8 cell treated with 1 M B-estradiol, 1 M progesterone, 1 M hydrocortisone, 1 M dexamethasone, 10 (-8) M calcitriol and 10 (-8) M alfacalcidol was decreased by 19%, 56%, 60%, 64%, 28% and 38% of control, respetively. Production of TGF-Bl by ROS 17/2.8 cells treated with B-estradiol, progesterone, hydrocortisone, dexamethasone, calcitriol and alfacalcidol was elevated by 30%, 18%, 60%, 48%, 405% and 452% of control, respectively. Alkaline phosphatase activity of ROS 17/2.8 cell treated with hydrocortisone, dexamethasone, calcitriol and alfacalcidol was increased by 3.4 times, 12.9 times, 10.0 times and 5.1 times of control, respectively, but with B-estradiol and progesterone decreased by 39% and 10% of control respectively. Northern blot experiments demonstrated that calcitriol and alfacalcidol increased mRNA levels of both osteocalcin and osteopontin.