Progress on the mechanism of PDGF-BB signaling pathway in the pulmonary vascular remodeling of neonatal hypoxic pulmonary hypertension / 国际儿科学杂志

Neonatal hypoxic pulmonary hypertension(HPH)is a common acute critical disease in NICU, and is one of the diseases leading to neonatal death.At present, the specific pathogenesis is still unclear.Current studies have shown that pulmonary vascular remodeling is an important pathological feature of pulmonary hypertension, and the excessive proliferation and migration of pulmonary artery smooth muscle cell is the main cause of pulmonary vascular remodeling.Platelet-derived growth factor(PDGF-BB)is a powerful mitogenic factor which involved in cell proliferation and migration.Currently, plenty of studies have found that PDGF-BB plays an important role in multiple diseases, including tumor, atherosclerosis, pulmonary hypertension and pulmonary fibrosis.In view of the mechanism of PDGF-BB, this article reviews the possible mechanism of PDGF-BB in pulmonary vascular remodeling with neonatal HPH, aiming to provide a new direction for the therapies of reversing pulmonary vascular remodeling with neonatal HPH.

Allogenic platelet-rich plasma in induced ulcers in rat's cornea / [Plasma rico em plaquetas alogênico em úlceras de córnea induzidas em ratos]

The objective in this study was to evaluate the clinic effect of applying allogenic platelet-rich plasma (PRP) heated or not, for treating cornea ulcers, including the dosage of PDGF-BB in the cornea. The ulcers were induced, standardizing the left eye from 81 rats (Ratus norvegicus, albinus variety), assigned randomly into three groups (N=27): control group (CG) which did not receive any topic treatment; heated PRP group (GA) and PRP group (GP), which received topical treatment every eight hours for five days. Each group underwent evaluation at 24 hours (M1), three days (M3) and five days (M5). The clinical exam evaluated the opacity, vascularization and corneal repair. The corneal PDGF-BB was dosed through the ELISA method. The corneal opacity was decreased in PRP-treated animals (GA and GP) and corneal repair time reduced when compared to CG at M1 and M5. Furthermore, GP showed greater vascularization at M3 compared to M1. Applied allogenic PRP eye drops, heated or not, speed up corneal healing, and reduce corneal repair time. However, the corneal PDGF concentration was not altered in any of the treatments.(AU)

Objetivou-se avaliar o efeito clínico da aplicação de plasma rico em plaquetas alogênico (PRP) aquecido ou não, no tratamento de úlceras de córnea, como a dosagem de PDGF-BB na córnea. As úlceras foram induzidas, padronizando-se o olho esquerdo de 81 ratos (Rattus norvegicus, variedade albinus), aleatoriamente, nos três grupos (N = 27): grupo controle (CG), que não recebeu nenhum tratamento tópico; grupo PRP aquecido (GA) e grupo PRP (GP), que receberam tratamento tópico a cada oito horas, durante cinco dias. Cada grupo foi subdividido em 24 horas (M1), três dias (M3) e cinco dias (M5). O exame clínico avaliou a opacidade, a vascularização e o reparo corneano. O PDGF-BB corneano foi dosado pelo método Elisa. Houve diminuição da opacidade da córnea nos animais tratados com PRP (GA e GP) e diminuição do tempo de reparo da córnea em comparação com CG, M1 e M5. Além disso, foi observada maior vascularização no GP no momento M3 em relação ao M1. A aplicação de colírios de PRP alogênico, aquecidos ou não, acelera a cicatrização da córnea, além de reduzir o tempo de reparo da córnea. No entanto, a concentração de PDGF na córnea não se alterou em nenhum dos tratamentos.(AU)

PDGF-BB promotes angiogenesis of human osteosarcoma through exosomes / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore whether PDGF-BB can be transmitted through exosome and verify its angiogenic function in human osteosarcoma. Methods: Exosomes from a variety of human osteosarcoma cells were isolated. The expression of PDGF-BB in cells and exosomes was detected by WB. Exosomes derived from osteosarcoma SJSA-1 cells were co-incubated with HUVEC, and the pattern of exosomal PDGF-BB entering HUVEC was observed using Immunofluorescence and confocal scanning microscope. SJSA-1 cell lines with PDGF-BB over-expression or knockdown were constructed by lentiviral infection, and the exosomes derived from transfected SJSA-1 cells were isolated and incubated with HUVEC. Microtubule formation experiment was conducted to detect their effects on angiogenesis; SJSA-1 cell transplanted xenograft model was established in nude mice, and the exosomes derived from SJSA-1 cells with PDGF-BB over-expression or knockdown were infused into nude mice to observe their effects on tumor growth. Results: The exosomes derived from osteosarcoma cells were successfully isolated, in which a large amount of PDGF-BB was confirmed. The exosomes entered HUVEC by endocytosis. The SJSA-1 cell lines with PDGF-BB over-expression or knockdown were successfully constructed, and the corresponding exosomes were isolated. Compared with the control group, exosomes with high PDGF-BB content significantly promoted HUVEC angiogenesis (P < 0.01 , t=13.51) and tumor growth (P < 0.01 ), while exosomes with low PDGF-BB content reduced the angiogenesis ability of HUVEC (P < 0.01 , t=8.226) and inhibited tumor growth (P < 0.01 ). Conclusion: The exosomal PDGF-BB secreted by osteosarcoma cells can be directly absorbed by HUVEC and induce tumor angiogenesis, further promoting the growth of osteosarcoma.

Expressions of TP53, PDGF and EGFR in Primary Astrocytomas and Their Correlation with Clinicopathological Features and Prognosis / 肿瘤防治研究

Objective To investigate the expression of TP53, PDGF and EGFR in primary astrocytomas, and analyze their correlation with clinicopathological features and prognosis. Methods We analyzed retrospectively the clinicopathological data of 90 patients with primary astrocytoma. The expressions of TP53, PDGF and EGFR in primary astrocytoma tissue samples were detected by immunohistochemistry. The survival of patients was followed up and Cox regression analysis was used to determine the prognostic factors. Results TP53 was expressed in the nucleus, PDGF and EGFR were expressed in the cytoplasm and cell membrane. The positive expression rates of TP53, PDGF and EGFR were 58.89%, 51.11% and 48.89%, significantly higher than those in normal brain tissues (all P < 0.05); the positive expression rate of TP53 in patients with tumor size ≥3 cm was higher than that in patients with tumor size < 3 cm. The positive expression rates of TP53, PDGF and EGFR in patients with WHO stages Ⅲ-Ⅳ were higher than those in patients with WHO stageⅠ-Ⅱ(all P < 0.05); the survival time of patients with positive expression of TP53, PDGF and EGFR were shorter than those of negative expression (all P < 0.05). Cox regression analysis found that WHO staging, TP53, PDGF and EGFR were all factors influencing the prognosis of primary astrocytomas patients. Conclusion TP53, PDGF and EGFR are highly expressed in primary astrocytomas and closely related to tumor progression. They are factors that affect the prognosis of patients.

Uso do hormônio do crescimento associado à fibrina rica em plaquetas e leucócitos injetável (I-PRF) / Use of the growth hormone associated with injectable platelet-rich fibrin (I-PRF)

Objetivo: demonstrar, por meio de uma revisão de literatura, a utilização do hormônio do crescimento (GH) e concentrados plaquetários e sugerir técnica de associação de uso para odontologia em processos de preservação de osso alveolar. Revisão de literatura: enxertos ósseos são uma necessidade na área da saúde, por diversas razões. A utilização de osso autógeno apresenta grande desvantagem em ter um segundo sítio cirúrgico, entretanto, os substitutos ósseos não possuem as características ideais. Assim, existe a busca por alternativas que otimizem a cicatrização e a incorporação dos substitutos ósseos, dentre elas os concentrados sanguíneos, ricos em fatores de crescimento derivados das plaquetas e o hormônio do crescimento. É possível encontrar uma vasta literatura utilizando os concentrados sanguíneos, inclusive utilizando esses como veículos para outras substâncias. Os concentrados sanguíneos são ricos em fatores de crescimento derivados das plaquetas, como fator de crescimento semelhante à insulina (IGF), Fator de crescimento derivado de plaquetas (PDGF) e outros. Além disso, também é possível encontrar, na literatura, o uso tópico de hormônio do crescimento em enxertos ósseos, fraturas e implantes dentários. Entretanto, o GH possui uma meia-vida de 20 minutos, assim, quando utilizado em conjunto com a I-PRF, espera-se um aumento no tempo de ação local. Considerações finais: é possível otimizar os enxertos ósseos utilizando-se L-PRF/I-PRF e hormônio do crescimento. Porém, são necessárias mais pesquisas.(AU)

Objective: this study aims to show through a literature review the use of the growth hormone and platelet concentrates and to suggest an association technique for dentistry use in alveolar bone preservation processes. Literature review: bone grafts are a health requirement for a number of reasons. The use of autogenous bone has the main disadvantage of a second surgical site, while bone substitutes do not present optimal characteristics. Thus, there is a search for alternatives that optimize the healing and incorporation of bone substitutes, which include blood concentrates that are rich in platelet-derived growth factors and the growth hormone. A vast literature can be found on blood concentrates, including their use as vehicles to other substances. Blood concentrates are rich in platelet-derived growth factors such as IGF, PDGF, and others. Moreover, the literature also shows the topical use of the growth hormone in bone grafts, fractures, and dental implants. However, the growth hormone presents a half-life of 20 minutes; therefore, when combined with I-PRF, an increased time in local action is expected. Final considerations: it is possible to optimize bone grafts by using L-PRF/I-PRF and the growth hormone. However, further research is required.(AU)

Role of periostin in PDGF-induced proliferation and extracellular matrix accumulation of mouse mesangial cells / 中国药理学通报

;Aim To investigate the role of periostin in PDGF-induced proliferation and extracellular matrix accumulation of mouse mesangial cells. Methods The effects of PDGF on periostin protein, cell proliferation and extracellular matrix accumulation were detected. The cells were collected at 0, 2,4, 6, 12 h after stimulation with PDGF(10 (ig • L " 1 ) to detect the expression of periostin, PCNA, FN and TGF-ßl by Western blot. The silencing effect of sh-periostin vector on periostin protein in mouse mesangial cells was identified by Western blot. Cells were randomly divided into control group, PDGF group, PDGF + sh-nc group and PDGF + sh-periostin group to detect the role of periostin in PDGF-induced proliferation and extracellular matrix accumulation of mouse mesangial cells. Results PDGF could elevate periostin protein expression. Western blot result showed that periostin protein expression in PDGF-stimulated groups was significantly higher than that in Oh group, which was consistent with the result of immunofluorescence. Positive expression of periostin was located in cytoplasm. Western blot result showed that PCNA, FN and TGF-ßl protein in PDGF-stimulated groups increased as compared with Oh group. shRNA vector aimed at periostin (sh-periostin vector) could partially reverse PDGF-induced mesangial cell proliferation and extracellular matrix expression. PCNA, Fn and TGF-ßl expressions were attenuated significantly. Conclusions PDGF can enhance periostin protein expression and increase mouse mesangial cell proliferation and extracellular matrix accumulation. Periostin shRNA vector can partially reverse PDGF-induced mesangial cell proliferation and extracellular matrix generation.

Effect of platelet-rich fibrin extract on the proliferation of gingival fibroblasts / 口腔疾病防治

Objective@#To study the effects of platelet-rich fibrin extract (PRFe) and platelet-derived growth factor (PDGF) released from PRFe on the proliferation of human gingival fibroblasts (HGFs) and to provide an experimental basis for its application in promoting gingival soft tissue increment.@*Methods@#Platelet-rich fibrin (PRF) was transformed into PRFe by tissue culture. The three-dimensional structure of PRF was observed by electron microscopy, and the content of PDGF in PRF was quantitatively determined by ELISA. The ratios of PRFe examined were 2.5% PRFe, 5% PRFe, 7.5% PRFe, 10% PRFe, 12.5% PRFe and 15% PRFe. Gingival fibrosis was detected by the CCK-8 method. After determining the optimal concentration of PRFe, flow cytometry was used to detect the effect of PRFe on the proliferation cycle of human gingival fibroblasts, and the effect of PDGF on the proliferative activity of gingival fibroblasts was observed by neutralizing the release of PDGF.@*Results @# PRF is a three-dimensional reticular structure that contains a large number of growth factors. PDGF release peaked on the 7th day. The proliferative activity of HGFs cultured with different concentrations of PRFe was concentration-dependent, but the effect was optimal at 5% PRFe (P < 0.05). There were no significant differences in the effect of subsequent concentration increases on the proliferation of HGFs (P > 0.05). The flow cytometry results showed that 5% PRFe could significantly stimulate the S-phase division and proliferation of gingival fibroblasts, while the PDGF neutralization test showed that the proliferation of gingival fibroblasts was significantly inhibited by the neutralization of PDGF.@*Conclusion@#Overall 5% PRFe had the best effect on promoting gingival fibroblast proliferation in vitro. PDGF released from PRF plays an important role in promoting the proliferation of gingival fibroblasts.

Effect and Mechanism of Guhong Injection Promoting Fracture Healing in Rats / 中国药学杂志

OBJECTIVE: To observe the effect of guhong injection(GHI) on tibial fracture healing in rats and to explore the mechanism of the action of GHI. METHODS: One hundred and eighty male SD rats were randomly divided into 6 groups with 30 rats in each group: sham operation group, model group, positive drug group(compound ossotide injection, 5 mL•kg-1), low, medium and high dose of GHI groups(2.5, 5, 10 mL•kg-1). In addition to the sham operation group, the other groups established the rat model of tibial fracture. All were given once daily intraperitoneal injections and samples were taken at 1st, 2nd, 4th and 6th week. Blood biochemical analysis and Elisa kit detection were performed on blood samples. X-rays, biomechanical tests, immunohistochemistry and RT-PCR were performed on the tibial samples. RESULTS: ①After administration for one, two and four weeks, the levels of serum calcium(Ca) and phosphorus(P) in medium and high dose of GHI groups were higher than those in model group(P<0.05). ②X-ray showed that the outer callus growth and the disappearance of fracture line in all dose groups of GHI were faster than those in model group. ③Compared with model group, the maximum load and rigidity of medium and high dose of GHI groups were increased at each time point(P<0.05), and the trend of stress line graph were improved obviously. The content of alkaline phosphatase(ALP) in medium and high dose of GHI groups were higher than that in model group at each time point(P<0.05). Compared with model group, the serum levels of PDGF were increased in all dose groups of GHI(P<0.05 or P<0.01). After administration for one, two and four weeks, the serum BMP-2 in all dose groups of GHI were higher than those in model group(P<0.05 or P<0.01). ⑤Compared with model group, the expression of Runx2 mRNA were increased in medium and high dose of GHI groups, as well as Smad5 protein expression(P<0.05 or P<0.01). CONCLUSION: GHI could significantly improve the biomechanical properties of bone in fracture rats. The promotion of fracture healing might be through the upregulation of PDGF and BMP-2 expression in different stages of bone healing, and the regulation of BMP/Smad5/ Runx2 signaling may be one of the mechanisms of promoting fracture healing.

PDGF-B immunogen preparation and the suppressive effect of anti-PDGF-B ascite antibody on the proliferation of hepG2 cells / 生物工程学报

To observe the immunogenicity of hPDGF-B immunogens that were synthesized with the fusional expression vector pET28-Trx and to test the suppressive effect of these specific antibodies induced by both of immunogens on proliferation of human HepG2 hepatoma cells. First, we chose 2 antigenic epitopes hPDGF-BΔ103-118aa and hPDGF-BΔ152-167aa from human PDGF-B and inserted these 2 coding regions into the empty vector plasmid pET28-Trx, separately. Second, mice were immunized with purified recombinant proteins to generate polyclonal antibody. Then we intraperitoneally injected mice bearing hepatoma 22 (H22) tumor cells to prepare antibody ascites. ELISA and Western blot were used to detect the titer and the utility of the antibody, respectively. Finally, HepG2 cells were exposed to PDGF-BB protein or anti-PDGF-B ascite antibody in different dilution concentrations groups and the proliferation of HepG2 cells was quantified by CCK8 assay. As the results, we identified mice that could produce high drop of neutralizing antibodies against hPDGF-B induced by both two recombinant proteins. Two anti-PDGF-B ascite antibodies could markedly inhibit the proliferation of HepG2 cells by blocking the stimulating effect of PDGF-BB protein. Our results suggest that Trx-PDGF-B recombinant protein as immunogen provides a new method for the preparation of PDGF-B vaccine, and also a new idea for the treatment of hepatocellular carcinoma in clinical practice.

Platelet-derived growth factor A mRNA in platelets is associated with the degree of hepatic fibrosis in chronic hepatitis C

Abstract: INTRODUCTION: Transforming growth factor beta 1 (TGFB1) and platelet-derived growth factor (PDGF) are the main cytokines related to hepatic fibrogenesis. METHODS: RNA isolated from the platelets and hepatic tissue of 43 HCV carriers was used for quantitative polymerase chain reaction to determine TGFB1, PDGFA, and PDGFB RNA expression. RESULTS: The mRNA expression of PDGFA in platelets was significantly lower in the group with advanced fibrosis than in the group with early-stage fibrosis. TGFB1 was more frequently expressed in platelets than in hepatic tissue, which was different from PDGFB. CONCLUSIONS: A pathway mediated by overexpression of TGFB1 via PDGFA in megakaryocytes could be involved in the development of fibrosis.

Efeitos do PDGF-BB na taxa de proliferação e na adesão de células derivadas da granulação óssea a fragmentos radiculares / Effects of PDGF-BB on the rate of proliferation and on the adhesion of human cells derived from bone granulation tissue to root fragments

O objetivo deste estudo foi investigar o papel do fator de crescimento derivado de plaquetas-BB (PDGF-BB) na concentração de 300ng/ml na taxa de proliferação e adesão de células derivadas da granulação óssea humana a fragmentos radiculares periodontalmente comprometidos. Na primeira etapa do estudo, foi estabelecida cultura primária de células da granulação óssea de dois pacientes adultos, sistemicamente saudáveis, não fumantes. Após a expansão celular, as células foram caracterizadas para determinação do fenótipo por meio de ensaios de viabilidade celular, MTT, ensaio de atividade de fosfatase alcalina, ensaio de mineralização e caracterização imunohistoquímica por meio de citometria de fluxo (segunda etapa). Na terceira etapa do estudo, os efeitos da adição de PDGF-BB recombinante humano na concentração de 300ng/ml na taxa de proliferação e adesão de células derivadas da granulação óssea a superfícies radiculares periodontalmente comprometidas foram investigados. A taxa de proliferação celular estimulada pelo PDGF-BB (grupo teste) ou pelo meio de cultura (grupo controle) foi investigada por meio de contagem de células viáveis nos frascos de cultura após 1, 3, 5 e 7 dias do cultivo celular. Foram obtidos 30 fragmentos dentários a partir de dentes extraídos por razões periodontais. Os fragmentos foram raspados com curetas Gracey e condicionados com solução em gel de EDTA a 24% durante 3 minutos, lavados com solução de soro fisiológico, secos e posicionados em placas de 24 poços. Foram incubadas sobre os fragmentos tratados 1x104 células GO por 24 horas, seguido por fixação e preparo para análise por microscopia eletrônica de varredura (MEV). O número de células aderidas sobre os fragmentos foi analisado nas fotomicrografias. O padrão de crescimento das células GO foi compatível com células ósseas, com modificação do padrão do crescimento com o aumento do número de passagens. Houve atividade de fosfatase alcalina em meio osteogênico e convencional, com pico máximo aos 7 dias e atividade de mineralização estimulada ou não por meio osteogênico, com pico máximo aos 21 dias. A análise por meio de citometria de fluxo demonstrou que as células GO não expressaram CD105 e CD166 na 14a passagem, indicando sua diferenciação celular avançada nesse período. A adição de rhPDGF-BB resultou em mudança na taxa de proliferação celular, observando-se pico máximo de crescimento aos 7 dias, com diferenças estatisticamente significantes (p < 0.005; ANOVA post hoc Tukey) em relação aos períodos de 1, 3 e 5 dias. O ensaio de MTT demonstrou maior viabilidade celular no período de 48 hs, comparativamente aos períodos de 24 e 72 horas, quando a densidade óptica celular diminuiu de forma significativa (p< 0.05; Friedmann pósteste Dunn). No ensaio de adesão celular, pode-se observar que a adição de rhPDGFBB aumentou significativamente o número de células aderidas aos fragmentos dentários (p< 0.05; teste t não pareado com correção Welch), com alteração da morfologia celular. Esses resultados sugerem que as células GO tem características compatíveis com linhagem de células osteoblásticas, de fenótipo mais diferenciado após a 12a passagem. A adição de rhPDGF-BB (300ng/ml) resulta em aumento da taxa de proliferação das células GO e do número de células aderidas a fragmentos radiculares, indicando que, nesta concentração, o fator de crescimento é citocompatível, favorecendo a proliferação e adesão celular.(AU)

The goal of this study was to investigate the effects of recombinant human platelet derived growth factor (rhPDGF-BB) at the concentration of 300ng/ml in the proliferation and adhesion of human bone granulation cells to periodontally diseased root fragments. At the first stage of the study, the granulation tissue existent in healing sockets (21 days after its creation) was collected from two systemically healthy nonsmoking adults to the establishment of primary culture. The in vitro properties of bone granulation (BG) cell lineage were characterized by cell viability, MTT, alkaline phosphatase activity and mineralization assays. The effects of culture medium (control) and rhPGDF-BB 300ng/ml (test) in the proliferation and adhesion of BG cells were investigated. The rate of BG cells proliferation was investigated by the number of viable cells present at 1, 3, 5 and 7 days after platting. Thirty root fragments were obtained from teeth extracted for periodontal reasons. Root fragments were scaled and root planed, conditioned with EDTA 24% for 3 minutes, rinsed in saline solution, air-dryed and positioned in 24-well plates. Each fragment was seeded with 104 BG cells, fixated after 24 hours and prepared for analysis in SEM. The number of cells adhered to the fragments was analysed in photomicrographies. BG cells growth pattern was compatible with osteogenic cell lineage, showing modification with the increasing number of cell passage. GO cells expressed alkaline phosphatase activity in conventional and osteogenic culture medium, with maximum peak at 7 days, as well as mineralization activity stimulated or not by osteogenic or non-osteogenic culture medium, with maximum peak at 21 days. The analysis by flow cytometer showed that BG cells have not expressed CD105 and CD106 at the 14th passage, indicating its advanced cell differentiation. The addition of rhPDGF-BB resulted in modification of proliferation rate, with maximum peak observed at 7 days, significantly different from 1-, 3- and 5-day periods (p< 0.005; ANOVA post hoc Tukey). MTT assay showed greater cell viability after 48 hours than after 24 and 72 hours, when optical density has significantly diminished (p< 0.05; Friedmann post hoc Dunn). At cell adhesion assay, it could be observed that the adhesion of rhPDGF-BB has significantly increased the number of cells adhered to root fragments (p< 0.05; unpaired t test with Welchs correction), and alterations in cell morphology. These results suggest that BG cells present in vitro characteristics compatible with osteoblastic cell lineages, with a more differentiated phenotype after the 12th passage. The addition of rhPDGF-BB (300 ng/ml) results in increase of the rate of BG cell proliferation and in the number of cells adhered to root fragments, indicating that, at this concentration, the growth factor is compatible with BG cells and favors cells proliferation and adhesion.(AU)

Effects of Hedysari Polysaccharide on Expressions of TSP-1 and PDGF-B in Retina of Diabetic Rats / 中国中医药信息杂志

Objective To observe the effects of Hedysari Polysaccharide (HPS) on the expressions of TSP-1 and PDGF-B in the retina of diabetic rats;To discuss the protective effect and possible mechanism on diabetic retinopathy. Methods The diabetic model was established by intraperitoneal injection of streptozotocin. 50 male SPF Wistar rats were randomly divided into 5 groups:model group, calcium dobesilate group, and HPS high-, medium-, and low-dose group, extra 10 rats were set as the normal group, 10 rats in each group. Each administration group was given relevant medicine for gavage, while model group and normal control group were given same amount NS for gavage, once a day for 8 weeks. The mRNA and protein expression of TSP-1 and PDGF-B were detected by qRT-PCR and immunohistochemistry. The retinal structure was observed by HE staining. Results HE staining showed that each layer of the retina of the model group was clear and complete, but the outer nucleus layer became looser, thinner and more disorderly, and the number of ganglion cells decreased slightly; the administration groups were improved markedly compared with the model group. Compared with the normal control group, the mRNA level and protein expression of retina TSP-1 on the model group dramatically dropped (P<0.01), and those of PDGF-B strikingly increased (P<0.01);Compared with the model group, the mRNA level and protein expression of retina TSP-1 on alladministration groups rose (P<0.05, P<0.01), and those of PDGF-B went down (P<0.01); Compared with all other administration groups, there was statistical significance in the mRNA level and protein expression of retina TSP-1 and PDGF-B on HPS high-dose group (P<0.05, P<0.01). Conclusion HPS may prevent the angiogenesis and proliferation in diabetic retinopathy process through adjusting the content of TSP-1 and PDGF-B in retina of diabetic rats so as to protect the retina.

Effects of conbercept combined with PDGF receptor inhibitor on the proliferation, migration of RPE cells and VEGF expression in hypoxic conditions / 眼科新进展

Objective To investigate the effects of conbercept combined with platelet-derived growth factor (PDGF) receptor inhibitor on the proliferation and migration of human retinal pigment epithelial cells (ARPE-19),as well as mRNA and protein expression of vascular endothelial growth factor (VEGF).Methods ARPE-19 cells were cultured in in vitro and the cells in logarithmic growth phase were obtained and induced by different concentrations of CoCl2.Then cell proliferation and toxicity test kit (CCK-8) was used to detect the proliferation activity of ARPE-19 cells for screening the optimal concentration of CoCl2 to construct hypoxic model.The cultured ARPE-19 cells were divided into normal group,hypoxic group and hypoxia + conbercept group (different concentrations),and CCK-8 assay was applied to analyze the effects of conbercept with different concentrations on cell proliferation activity for screening the best concentration of conbercept.The ARPE-19 cells in logarithmic phase were divided into normal group,hypoxic group,hypoxia + PDGF receptor inhibitor group (different concentrations),and CCK-8 assay was performed to detect the effects of PDGF receptor inhibitor on cell proliferation with hypoxic damage activity for screening the best concentration of PDGF receptor inhibitor.The logarithmic growth phase cells were divided into normal group,hypoxic group,hypoxia + 20 mg · L-1 conbercept group,hypoxia + 200 μmol · L-1 PDGF receptor inhibitor group,hypoxia + conbercept + PDGF receptor inhibitor group (both optimal concentration),and then the cell proliferation in each group was detected by CCK-8 assay,and the migration was detected by transwell chambers.The level of VEGF protein in the supernatant of each group was detected by ELISA methods,and VEGF mRNA expression was measured by RT-PCR.Results CCK-8 assay results showed that the A value of ARPE-19 in the 100 μmol · L-1 CoCl2 group was the highest (1.063 ± 0.031),which was set to be the optimal concentration for preparation of hypoxic model.CCK-8 assay results showed the cell proliferation rate of 20 μg · mL-1 conbercept and 20 μmol · L-1 PDGF receptor inhibitor group was (94.58 ± 3.80) ％ and (96.72 ± 5.44) ％,respectively,which could achieve the most satisfying efficacy,thereby both concentrations were selected for follow-up experiments.The cell proliferation and migration ability and VEGF protein level decreased in the conbercept group,PDGF receptor inhibitor group and conbercept + PDGF receptor inhibitor group when compared with the hypoxic group,and the decrease in the combined group was the most significant.Moreover,VEGF mRNA expression in the conbercept group and combined group were decreased when compared with the hypoxic group.Conclusion Hypoxia can enhance cell proliferation and migration ability and induce the up-regulation of VEGC protein and mRNA expression.In addition,PDGF receptor inhibitor combined with conbercept has inhibitory effects on the migration and proliferation of ARPE-19 cells as well as VEGF protein and mRNA expression following hypoxia injury,which is superior to conbercept treatment alone.

AngⅡ promotes liver fibrosis through PDGF signal pathway / 实用医学杂志

Objective To investigate the effect and mechanism of AngⅡ on collagen in hepatic stellate cell. Methods HSCs were isolated and cultured, 3H-pro incorporation method was used to evaluate the effects of different doses of AngⅡ on the proline syntheses. RT-PCR assay were used to assess changes in mRNA expression levels of type Ⅰ and Ⅲ procollagen. PDGFR-β mRNA and protein were determined by in situ hybridization and immunocytochemistry. Results 10-8~ 10-5 mol/L AngⅡ could significantly increase the 3H-pro incorporation rate of HSC in a dose-dependent style, 10-6 mol/L AngⅡis the most effective dose. The cultured HSC showed a little expression of type Ⅰ and Ⅲ procollagen mRNAs, while 10-6 mol/L AngⅡwas able to enhance the expression for type Ⅰ and Ⅲ procollagen mRNAs significantly(P < 0.01). AngⅡalso could enhance both mRNA and protein expression of PDGFR-β on HSC(P < 0.01). Conclusion These results suggest that AngⅡ could promote HSC collagen synthesis by enhancing the expressions of PDGFR-β.

In vitro culture and migration assay of mouse smooth muscle progenitor cells / 中国药理学通报

Aim To establish a reliable method for the culture of mouse smooth muscle progenitor cells and in-vestigate their migration ability .Methods Mesenchy-mal stem cells from compact bones were obtained from C57 BL/6 mice and stimulated with PDGF-BB to in-duce these cells to differentiate into smooth muscle pro-genitor cells. Morphological analysis , immunocyto-chemical and flow cytometric analysis were used to i-dentify the cell type and the migration ability was in-vestigated by the transwell system and flow cytometry . Result After PDGF-BB stimulation for 7 days, the cells showed spindle shape and started to express α-SMA as demonstrated by immunocytochemistry .After 21 days induction , Flow cytometric analysis revealed that over 70%of the cells expressed both CD 34 andα-SMA and 58.5%of the cells expressed SM-MHC.Mi-gration assay showed that the smooth muscle progenitor cells from culture could migrate in vivo and in vitro. Conclusions The culture of smooth muscle progenitor cells from compact bone-derived mesenchymal stem cells is easily operated with high yield rate and shorten culture period . Obtained smooth muscle progenitor cells from culture could migrate in vivo and in vitro, which is suitable for the mechanism studies .

Effect of decitabine on serum platelet derived growth factor levels in patients with leukemia / 中国生化药物杂志

Objective To investigate the effect of decitabine on serum platelet derived growth factor levels in patients with leukemia. Methods 120 cases of acute myeloid leukemia patients admitted to our hospital from January 2012 to December 2014 were selected as the study subjects.The control group was given CAG regimen: 1-14 d Seventh days of intravenous infusion of 20 mg/d aclarubicin, first to fourteen days subcutaneous injection of 10 mg/m2 , 1 times per 12 h, 1-13 d cytosine arabinosine days under the subcutaneous injection of granulocyte colony stimulating factor 300μg/d.The observation group was given to the west of the lake 25 mg/d, intravenous infusion, used for 5 days.The levels of platelet derived growth factor PDGF in the two groups were compared and the therapeutic effect.Results Before treatment, differences of PDGF in control group and observation group was no statistical significance.After treatment, PDGF in observation group was lower than control group ( P <0.05).The proportion of primary cells of bone marrow of two groups after treatment was lower than before treatment, and the observation group was more lower.Total effective rate of observation group was 55.0%,which was higher than that in control group(35.0%)(P<0.05).Conclusion Decitabine could reduce the level of of PDGF in patients with leukemia, and improve the total effective rate.

The effects of PRF and its three growth factors on the proliferation and adhesion of rat ADSCs / 实用口腔医学杂志

Objective:To study the effects of PRF and recombinant hPDGF-AB,TGF-β1 and VEGF on the proliferation and adhe-sion of rat adipose tissue-derived stem cells(ADSCs)in vitro.Methods:ADSCs were cultured with PRF membrane and various do-ses of PDGF-AB,TGF-β1 and VEGF,cell adhesion was examined by adhesion assay after 2 culture,cell proliferation was examined by CCK-8 kit after 1 -7 d culture.Results:Cell adhesion assay showed that the adhesive numbers of rat ADSCs in PRF group were significantly higher than those in the negative group(P 0.05).The adhesive numbers of the ADSCs treated by VEGF or TGF-β1 at different concentrations showed significant difference(P <0.05).CCK-8 kit assay showed that at different time points, the A values of ADSCs in PRF group were significantly higher than those of the negative control group(P <0.05).The A values of ADSCs in VEGF or PDGF-AB groups at different concentrations showed significant difference(P <0.05).The A values of rat AD-SCs in TGF-β1 group at different concentrations were lower than those in the negative control group(P <0.05).Conclusion:PRF as a combination of growth factors may stimulate the proliferation and adhesion of rat ADSCs in vitro.PDGF-AB and VEGF may stim-ulate the proliferation of rat ADSCs.TGF-β1 and VEGF may stimulate the adhesion of rat ADSCs in a dose-response manner to some degree.

Estimation of the use of fibrin and collagen membranes as carriers for platelet-derived growth factor-BB (PDGF-BB) in the presence of amoxicillin.

The effect of homogeneous fibrin (Fb), collagen (Coll) and composite fibrin-heparin (Fb-Hp), fibrin-collagen (Fb-Coll) membranes on in vitro release of platelet-derived growth factor (PDGF-BB) was evaluated in the presence or absence of amoxicillin using of the ELISA immunoassay test. Amoxicillin concentration was determined spectrophotometrically at 272 nm. The process of the PDGF-BB growth factor and amoxicillin release from the studied membranes was of a two-phase nature in the majority of the systems analysed. The PDGF-BB was released in the highest amount from the Coll membrane (M7) without the presence of amoxicillin – 546.2 ±7.47 pg, t0.5 = 0.88 h and 202.5 ± 6.83 pg, t0.5 = 26.65 h during the first phase and second phase, respectively. The lowest PDGF-BB release was observed from composite M4 (Fb-Hp) membrane – 5.88 ± 0.81 pg, t0.5 = 1.69 h; and 110.2 ± 6.48 pg, t0.5 = 855.6 h during first and second phase respectively. An optimal release of amoxicillin was observed in the case of the composite M6 (Fb-Coll) membrane – only in the second phase: 64.2 ± 7.8 mg, t0.5 = 83.5 h. The lowest and delayed amoxicillin release was achieved for M4 membrane (approx. 17.1 ± 1.12 mg, t0.5 = 46.5 h). The results of the PDGF-BB release and amoxicillin from membranes indicated a correlation between the level of release and composition of the film. Our results suggested that fibrin and collagen membranes may be beneficial to enhance periodontal bone regeneration.

Effect of PDGF-Rb antagonist imatinib on endometrial injury repairing in mouse model

OBJECTIVE@#To study the effects of PDGF-Rb antagonists imatinib on endometrial injury repairing in the mouse model.@*METHODS@#The cultured MSCs cells from male mice were marked with BrdU in vitro, and then transplanted to the female mice which suffered from radiation injury through tail vein, PDGF-Rb antagonists imatinib was injected through abdominal cavity. Four groups were arranged, which were radiation transplantation group, normal control group, imatinib intervention group and radiation control group. BrdU incorporation, SRY expression and MVD status were detected in uterus of mice.@*RESULTS@#SRY gene was negative expressed in normal control group and radiation control group. SRY gene presented positive in radiation transplantation group and imatinib intervention group; BrdU incorporation showed negative in radiation control group and normal control group which died in the early stage in mice; the incorporation of BrdU was higher in radiation transplantation group compared with imatinib intervention group; CD34 was positive on the uterus of all the four groups, which showed highest in radiation control group and lowest in radiation control group; The MVD in imatinib intervention group was lower than radiation control group; the difference of MVD was significantly compared with normal control group (P < 0.05).@*CONCLUSIONS@#PDGF-Rb antagonists imatinib could inhibit the repairing function of MSCs in the endometrial lesions in mice.

Effect of PDGF-Rb antagonist imatinib on endometrial injury repairing in mouse model

Objective: To study the effects of PDGF-Rb antagonists imatinib on endometrial injury repairing in the mouse model. Methods: The cultured MSCs cells from male mice were marked with BrdU in vitro, and then transplanted to the female mice which suffered from radiation injury through tail vein, PDGF-Rb antagonists imatinib was injected through abdominal cavity. Four groups were arranged, which were radiation transplantation group, normal control group, imatinib intervention group and radiation control group. BrdU incorporation, SRY expression and MVD status were detected in uterus of mice. Results: SRY gene was negative expressed in normal control group and radiation control group. SRY gene presented positive in radiation transplantation group and imatinib intervention group; BrdU incorporation showed negative in radiation control group and normal control group which died in the early stage in mice; the incorporation of BrdU was higher in radiation transplantation group compared with imatinib intervention group; CD34 was positive on the uterus of all the four groups, which showed highest in radiation control group and lowest in radiation control group; The MVD in imatinib intervention group was lower than radiation control group; the difference of MVD was significantly compared with normal control group (P < 0.05). Conclusions: PDGF-Rb antagonists imatinib could inhibit the repairing function of MSCs in the endometrial lesions in mice.