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Endogenous pigment epithelium derived factor (PEDF) shows great potential as a drug target for the treatment of diabetes retinopathy (DR) due to its anti-angiogenesis, anti-inflammatory, neuroprotective and neurotrophic effects. PEDF plays a biological role by combining with receptor proteins on cell membrane surface and regulating a variety of signaling pathways. Low density lipoprotein receptor related protein 6 plays a role in inhibiting oxidative stress reaction, inflammatory reaction, and neovascularization of DR. Adipose triglyceride lipase, laminin receptor, plexin domain containing 1 (PLXDC) 1, PLXDC2 and F 1-adenosine triphosphate synthase have the effect of promoting endothelial cell apoptosis, among which PLXDC1 also has neuroprotective effect. By clarifying the receptor that PEDF acts on, exploring the affinity between the receptor and PEDF, the difference in the expression level of each receptor in the process of disease, and the specific function that PEDF plays after binding with specific receptors, we can develop fusion protein drugs for the active domain of high affinity of receptors, have a clearer understanding of the pathogenesis of DR, and take PEDF or PEDF receptor as the target to consolidate the theoretical basis for the development of new therapeutic drugs and strategies for DR.
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Objective:To explore the changes in morphology and function of meibomian gland and the expressions of inflammatory factors and lipid metabolic factors in meibomian gland of diabetic mice.Methods:Fifty 8-week-old male C57BL/6 mice of clean degree were divided into normal control group ( n=20) and diabetes model group ( n=30) according to a random table.Diabetes model was established by the intraperitoneal injection of streptozotocin (60 mg/kg, 10 mg/ml). Mouse tail vein blood glucose ≥16.7 mmol/L was considered as successful modeling.Blood glucose was measured weekly, and body weight was compared between the two groups.Ten mice were randomly selected for fluorescein sodium staining of the cornea to evaluate the integrity of the corneal epithelium from both groups at an interval of 4 weeks.Five mice were randomly selected from the two groups and were sacrificed via anesthesia to collect meibomian gland tissue for hematoxylin and eosin staining in order to observe morphological changes at 8 and 16 weeks after modeling, respectively.At 16 weeks following modeling, mebomian gland of 5 mice randomly selected from both groups was stained with oil red O staining to observe the distribution of lipid.Real-time fluorescence quantitative-PCR was performed to detect the relative expressions of tumor necrosis factor (TNF)-α, pigment epithelium derived factor (PEDF), peroxisome proliferators-activated receptor γ (PPARγ), and adipose differentiation-related protein (ADFP) mRNA in meibomian gland.The use and care of animals complied with the ARVO statement.This study protocol was approved by the Institutional Animal Care and Use Committee of Tianjin Medical University Eye Hospital (No.TJYY20190630009). Results:The successful modeling rate of diabetes in mice was 100%, and the survival rate was 83.3% (25/30). The weight was significantly lower and the blood glucose level was higher in diabetes model group at 8 and 16 weeks after modeling in comparison with normal control group (all at P<0.05). There were significant differences in corneal fluorescein staining score among different time points in diabetes model group ( F=27.155, P<0.05). In diabetes model group, thinner wall of meibomian gland duct, enlarged lumen of the duct, dilated acini and oil red-stained lipid deposition in most acini were observed.At 16 weeks after modeling, the expressions of TNF-α, and PPARγ mRNA in meibomian gland of diabetes model group were 3.33±0.91 and 1.55±0.25, which were significantly higher than 1.00±0.16 and 1.00±0.27 of normal control group (both at P<0.05). The expression of PEDF mRNA in diabetes model group was 0.42±0.08, which was significantly lower than 1.00±0.34 in normal control group ( P<0.05). There was no significant difference in the ADFP mRNA expression between the two groups ( t=0.943, P=0.38). Conclusions:Inflammatory factors and lipid metabolic factors such as TNF-α, PEDF, and PPARγ may be involved in the pathogenesis of meibomian gland dysfunction induced by diabetes.
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Objective:To investigate the protein expression changes of human umbilical cord mesenchymal stem cells (hUCMSCs) modified with pigment epithelium-derived factor ( PEDF) gene mediated by lentivirus. Methods:The hUCMSCs were divided into the control group and experimental group.Cells in the control group were normal hUCMSCs and the cells in experimental group were hUCMSCs with PEDF modification.The proteins from the two groups were collected and processed by FASP method.Samples were fractionated by liquid chromatography and analyzed by tandem mass spectrometry, and SWATH mode was applied.Differential protein analysis, Gene Ontology (GO) analysis and Reactome pathway enrichment analysis were performed. Results:A total of 5 361 quantified proteins were detected in this experiment, of which 432 proteins were differentially expressed (fold change>1.5, P<0.05). There were 219 of the 432 proteins up-regulated, including serpin family F member 1 (SERPINF1) (PEDF), DEAD-box helicase 59 (DDX59) and thrombospondin 1 (THBS1), etc., whereas 213 proteins were down-regulated, including collagen type Ⅰ alpha 1 (COL1A1), COL18A1, etc.GO analysis indicated that the differential proteins were mainly involved in fibrinolysis, extracellular structure organization, regulation of transporter activity, biosynthetic process of secondary alcohol and cholesterol, coenzyme metabolic process and regulation of peptidase activity, etc.Reactome pathway analysis showed that the differential proteins were mostly involved in regulation of insulin like growth factor (IGF) transport and uptake by IGF binding protein, post-translational protein phosphorylation, extracellular matrix organization, metabolism of steroids. Conclusions:After gene modification with PEDF mediated by lentivirus, the expression of many proteins in hUCMSCs were changed. PEDF gene modification can alter the structure of extracellular matrix and regulate the expression of proteins associated with cell proliferation, self-renewal and multipotency.
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@#AIM: To explore the application value of serum pigment epithelial derived factors(PEDF), fibroblast growth factor 19(FGF19)and β-endorphin(β-EP)in the diagnosis and severity assessment of primary glaucoma. <p>METHODS: A total of 102 patients with primary glaucoma in the hospital were enrolled as study group between February 2018 and February 2020, while other 102 healthy controls during the same period were enrolled as control group. The levels of peripheral serum PEDF, FGF19 and β-EP were compared between the two groups. And their diagnostic value for primary glaucoma was analyzed. The study group was divided into severe and non-severe groups according to the diagnostic criteria for severe primary glaucoma. The levels of peripheral serum PEDF, FGF19 and β-EP were compared between severe group and non-severe group. And their evaluation value for disease severity was analyzed. The risk factors of disease severity were analyzed by multivariate Logistic regression analysis. <p>RESULTS: The level of serum PEDF in study group was significantly lower than that in control group, while levels of FGF19 and β-EP were significantly higher than those in control group(<i>P</i><0.001). AUC values of PEDF, FGF19 and β-EP levels in the diagnosis of primary glaucoma were 0.695, 0.754 and 0.768, respectively. The level of serum PEDF in severe group was significantly lower than that in non-severe group(<i>P</i><0.001), while levels of FGF19 and β-EP were significantly higher than those in non-severe group(<i>P</i><0.001). <i>AUC</i>values of PEDF, FGF19 and β-EP levels in assessing the severity of primary glaucoma were 0.731, 0.709 and 0.685, respectively. PEDF lower than 9.66pg/mL, FGF19 higher than 143.75ng/L and β-EP higher than 106.27ng/L were independent influencing factors of severe primary glaucoma(<i>OR</i>=2.280, 1.570, 1.413, all <i>P</i><0.05). <p>CONCLUSION: FGF19 and β-EP are of auxiliary diagnostic value in primary glaucoma, while PEDF and FGF19 are of evaluation value in disease severity. PEDF lower than 9.66pg/mL, FGF19 higher than 143.75ng/L and β-EP higher than 106.27ng/L are independent influencing factors of severe primary glaucoma.
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BACKGROUND@#The anti-tumor effect of pigment epithelium-derived factor (PEDF) has been widely confirmed. However, the anti-tumor effect of its peptides is rarely reported. This study aims to investigate the effects of PEDF and its peptides on the apoptosis and migration of non-small cell lung cancer (NSCLC).@*METHODS@#In this study, A549 cells and H1299 cells were selected as the research object, and the cells were divided into normal group, PEDF treatment group, 34 peptide treatment group, 44 peptide treatment group and 34+44 peptide treatment group by administering different drugs at the same concentration to the cells. The proliferation activity of cells in each group was detected by CCK-8 method; the migration ability of cells was detected by scratch test; the expression levels of apoptosis related proteins such as protein kinase 3 (RIP3) and cleaved-caspase-3 were detected by Western blot; the expression levels of epithelial mesenchymal transition (EMT) markers in each group, such as cadherin (E-cadherin) and α-smooth muscle actin (α-SMA) were detected by Western blot; the apoptosis rate of each group was detected by flow cytometry.@*RESULTS@#The results of CCK-8 showed that PEDF and its peptides could inhibit cell proliferation, and the inhibitory effect of 34+44 peptide was the strongest (P<0.05); Observation under the microscope found that PEDF and its peptides can inhibit the proliferation and mesenchymal transformation of A549 cells and H1299 cells, and the inhibitory effect of the 34+44 peptide group is the most obvious; Western blot indicated that compared with other groups, the expressions of cleaved-caspase-3 and RIP3 in 34+44 peptide group were significantly higher (P<0.05), and the expressions of EMT protein E-cadherin were higher, the expression of α-SMA decreased (P<0.05); The results of flow cytometry showed that the apoptosis rate of 34+44 peptide group was significantly higher than those of other groups (P<0.05); The scratch test showed that compared with all the other groups, the healing rate of 34+44 peptide group was the lowest (P<0.05).@*CONCLUSIONS@#34+44 combination peptide can better promote the apoptosis of NSCLC, inhibit the migration of NSCLC, and thereby inhibit the growth of NSCLC.
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Humans , Apoptosis , Cadherins/genetics , Carcinoma, Non-Small-Cell Lung/genetics , Caspase 3 , Cell Line, Tumor , Cell Movement , Cell Proliferation , Epithelial-Mesenchymal Transition , Eye Proteins , Lung Neoplasms/genetics , Nerve Growth Factors , Peptides/pharmacology , Serpins , SincalideABSTRACT
@#AIM: To quantitatively measure and evaluate the VEGF-A, platelet derived growth factor(PDGF)and pigment epithelium derired factor(PEDF)in the aqueous humor of patients with neovascular glaucoma(NVG). <p>METHODS: Prospectively clinical study. This study involved 23 eyes of 23 patients with advanced NVG and 23 control subjects with age related cataract. Protein concentrations of VEGF-A, PDGF and PEDF in aqueous humor and plasma were determined by enzyme-linked immunosorbent assay(ELISA)tests. <p>RESULTS: The VEGF-A and PDGF concentrations in aqueous humor from NVG patients were(1130.56±69.32)ng/L and(221.95±56.08)ng/L, respectively. Both of them were significantly higher than control subject(226.45±37.46)ng/L,(36.25±7.12)ng/L(<i>P</i><0.01). Aqueous PEDF was significantly lower in the NVG group(195.69±42.00)ng/L than that in controls(497.89±12.52)ng/L(<i>P</i><0.01). However, levels of VEGF-A, PDGF and PEDF in the serum of NVG were(226.45±37.46)ng/L,(29.57±6.31)ng/L and(13.24±1.76)ng/L, respectively, which were similar with control subjects(219±34.89)ng/L,(28.28±7.24)ng/L and(12.96±2.08)ng/L(<i>P</i>>0.05). The concentrations of VEGF-A were closely positive correlated with levels of PDGF in the aqueous humor of patients with NVG(<i>r</i>=0.502, <i>P</i>=0.015). However, the concentrations of VEGF-A were closely negative correlated with levels of PEDF in the aqueous humor of patients with NVG(<i>r</i>=-0.480, <i>P</i>=0.020). <p>CONCLUSION: There were higher levels of VEGF-A and PDGF, and lower level of PEDF in the aqueous humor of patients with NVG. There was a positive correlation between VEGF-A and PDGF, a negative correlation between VEGF-A and PEDF. The combination of anti-VEGF agent, PDGF inhibitor and PEDF may provide a new idea for the treatment of NVG.
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Objective To observe the expression of FGF19 (fibroblast growth factor 19) and PEDF (pigment epithelium-derived factor) in the serum of type 2 diabetic retinopathy (DR) patients and discuss their significance.Methods Total 89 patients with type 2 diabetes were selected and divided into two groups according to whether they were combined with diabetic retinopathy:45 patients with type 2 diabetes alone (DM group) and 44 patients with type 2 diabetes mellitus combined with retinopathy (DR group).At the same time,40 healthy people were selected as the control group (NDM group).The serum levels of FGF19 and PEDF and their biochemical indexes were detected and compared in each group.The correlation between the indexes and the relationship between FGF19,PEDF and DR were analyzed.Results Compared with NDM group,serum FGF19 level in DM group and DR group decreased,while C-reactive protein (CRP) and PEDF level in DM group and DR group increased (P ＜ 0.05);serum FGF19 level in DR group was lower than that inDMgroup,while serum PEDF level was higher than that in DM group (P ＜0.05).The levels of fasting blood glucose (FBG),fasting insulin levels (FINS),glycosylated hemoglobin (HbA1c),total cholesterol (TC),high density lipoprotein (HDL) and low density lipoprotein (LDL) in DM and DR groups were higher than those in NDM group (P ＜ 0.05),but there was no significant difference between DM group and DR group (P ＞ 0.05);the levels of Hcy,triacylglycerol (TG) and CRP in DR group were higher than those in DM and NDM group (P ＜ 0.05),and the levels of VLDL were lower than those in DM and NDM group (P ＜ 0.05).The level of serum FGF19 was positively correlated with HDL and CRP in patients with DR (r =0.341,0.623,P ＜ 0.05),and negatively correlated with fasting blood glucose(r =-0.428,P ＜0.05);The level of serum PEDF in patients with DR were negatively correlated with FINS (r =-0.343,P ＜0.05).When the serum level of PEDF ＞ 12.76 mg/L,the sensitivity and specificity of PEDF dignosing DR were 80.9％ and 56.7％ respectively.Conclusions In patients with DR,serum FGF19 level decreased and serum PEDF levels increased.The level of both changes is closely related to DR and may be involved in the development of DR.
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Objective To explore the role of pigment epithelium-derived factor (PEDF) in vascular endothelial cells dysfunction induced by sodium arsenite (NaAsO2).Methods Human umbilical vein endothelial cells (EA.Hy926 cells) were treated with different levels of NaAsO2 [0 (control),1,2,5,10,20,50 μmol/L] for 24 hours.The cell viability was determined using CCK8.Colorimetric assay was used to detect the activity of inducible nitric oxide synthase (iNOS) in culture supematants,PEDF content in the supematant of EA.Hy926 cells was detected by enzyme-linked immunosorbent assay (ELISA),and nitric oxide (NO) content in cells was detected by flow cytometry.Results Compared with the control group [(101.08 ± 3.22)％],the cell viability of 20 μ mol/L group [(80.69 ± 7.95)％] and 50 μmol/L group [(69.87 ± 10.54)％] decreased significantly,and the differences were statistically significant (P ＜ 0.05).The activity of iNOS increased significantly in 10,20,50 μmol/L groups [(829.1 ± 68.2),(772.3 ± 37.1),(874.6 ± 43.5) U/L],respectively,in comparison with that of the control group [(397.5 ± 43.5) U/L] in the cell culture supematant (P ＜ 0.01).Similarly,PEDF levels in the groups of 10,20,50 μmol/L [(12.06 ± 0.55),(11.97 ± 0.39) and (13.89 ± 0.26) mg/L respectively] were higher than that of the control group [(10.70 ± 0.35) mg/L,P ＜ 0.01],and the highest content of PEDF was found in 50 μmol/L group.The NO level in 50 μmol/L group (11 558.99 ± 397.43) was significantly lower than that of the control group (14 131.49± 262.61,P ＜ 0.01).Conclusion PEDF is involved in the vascular endothelial cells dysfunction induced by arsenic.
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@#AIM: To observe the effect of pigment epithelium-derived factor(PEDF)on retinal neovascularization(RNV)and monocyte chemoattractant protein-1(MCP-1)expressions in mice retina of oxygen-induced retinopathy(OIR), and to investigate the protective effect of PEDF on ischemia hypoxia retinopathy and the possible mechanism. <p>METHODS: A total of 160 postnatal day(P)7 C57BL/6 mice were obtained. All mice except normal control group were exposed to(75±2)% oxygen environment for 5d and then kept in room air for another 5d to establish the OIR mice model. All mice in normal control group(40 mice)were exposed to room air only. At P12 and P14, respectively, mice in PEDF treatment group were injected intravitreously with recombinant human PEDF(2μg/eye,1μL)in the right eye, while mice in treatment control group were injected intravitreously with the same volume of vehicle [1μL, 10mmol/L phosphate buffered saline(PH7.4), PBS] in the right eye. All mice were euthanized at P17. Eyes were whole mounted and stained with Lectin to observe the growth of abnormal RNV; And retinal specimens were prepared for PEDF, MCP-1 protein and mRNA analysis by Western blot and real time RT-PCR respectively. <p>RESULTS: Changes of retinal vessels had been detected by fluorescence microscopy on flat-mounted retina. The relative RNV areas were significantly increased in OIR model group compared with those in normal control group(<i>P</i><0.01). However, the relative RNV areas were significantly reduced in PEDF treatment group compared with those in PBS treatment control group(<i>P</i><0.01). The specific expression of MCP-1 protein and mRNA in the OIR model group were higher than those of normal control group, presenting a statistically significance(both <i>P</i><0.05). The specific expression of PEDF protein and mRNA in the OIR model group showed a considerable decline in comparison with normal control group, presenting a statistically significance(both <i>P</i><0.01). And the specific expression of MCP-1 protein and mRNA in those of PEDF-treated group showed a considerable decline in comparison with PBS-treated group, and the differences were statistically significant(both <i>P</i><0.05). However, there were increase of the expression of MCP-1 protein and mRNA between normal control group and PEDF-treated group, presenting no statistically significance(both <i>P</i>>0.05). <p>CONCLUSION: PEDF could inhibit oxygen-induced retinal neovascularization and down-regulate retinal MCP-1 expression under hypoxia, which may underlie its anti-neovascularization effects and play a role of protection in ischemic retinopathy.
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Objective Studies are rarely reported on the effect of short peptides of the pigment epithelium derived factor (PEDF) on the proliferation of human cutaneous squamous (SCL-1) cells. The purpose of this study is to investigate segmented cloning and expression of the PEDF protein and observe its effect on the proliferation of human SCL-1 cells. Methods The target genes of PEDF1, PEDF2 and PEDF3 were amplified by PCR and the recovered fragments subjected to double digestion of NheⅠ and Hind Ⅲ and inserted into the pET28a(+) plasmid. The product was transformed into human E coli BL21 and induced to express, followed by isolation and purification of the fusion protein. CCK-8 assay was used to detect the proliferation of the SCL-1 cells with PEDF1, PEDF2 and PEDF3 at 100, 400, 800 and 1000 nmol/L at 24, 48 and 72 hours. Results The prokaryotic expression vectors of PEDF1, PEDF2 and PEDF3 were successfully constructed, and their fusion proteins prepared, with the molecular weight of 18 000, 17 000 and 13 000, respectively. The proliferation of the SCL-1 cells was significantly decreased in the 800 and 1000 nmol/L PEDF3 groups compared with that in the 0 nmol/L PEDF3 group at 24 hours (0.16 ± 0.03 and 0.78 ± 0.07 vs 1.00 ± 0.00, P < 0.05), inhibited in a concentration-dependent manner in the 400, 800 and 1000 nmol/L PEDF3 groups at 48 hours (P < 0.05), markedly lower in the 800 and 1000 nmol/L PEDF3 groups at 72 hours (0.53 ± 0.05 and 0.51 ± 0.05) than in the in the 400 and 0 nmol/L PEDF3 groups (0.60 ± 0.05 and 1.00 ± 0.00) (P < 0.05). Conclusion The PEDF fusion proteins were successfully segmentally cloned and expressed and PEDF3 inhibited the proliferation of SCL-1 cells, which has paved the ground for further screening of active functional short peptides of PEDF.
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AIM: To observe the clinical efficacy of different treatment modalities and the influence on the levels of pigment epithelium-derived factor ( PEDF ) and vascular endothelial growth factor ( VEGF ) in patients with neovascular glaucoma ( NVG) in high altitude area.?METHODS: Ninety cases of patients ( 90 eyes ) with NVG treated in our hospital were selected as the study objects, and they were divided into Group A and Group B according to different surgery methods, 45 cases in each groups. Group A was given Ex-press glaucoma drainage device implantation, and Group B was given trabeculectomy combined with cyclocryotherapy. We observed the clinical efficacy, intraocular pressure and visual acuity in the two groups, detected the PEDF and VEGF levels in the aqueous humor, and recorded the postoperative complications.?RESULTS:The total effective rate of Group A was 91％, significantly higher than that of Group B 76％ (P<0. 05). At 2wk, 2 and 4mo after surgery, IOP levels were significantly lower than that before surgery (P<0. 05), and those in Group A were significantly lower than those in Group B (P<0. 05). There was no significantly difference in IOP levels at 6mo after surgery between the two groups (P>0. 05). At 1wk after operation, the incidence of visual reduction in Group A was 7％, significantly lower than that in Group B 22％ (P<0. 05). At 1wk after operation, PEDF levels were significantly higher than those before operation in the two groups (P<0. 05), and that in Group A was significantly higher than that in Group B (P<0. 05);VEGF levels were significantly lower than that before operation in the two groups (P<0. 05), and that in Group A was significantly lower than that in Group B (P<0. 05). The incidence rate of complications in Group A was 4％, significantly lower than that of 18％ in Group B at 6mo postoperatively (P<0. 05).?CONCLUSION: Ex- press glaucoma drainage device implantation, trabeculectomy combined with cyclocryotherapy can both be used to treat patients with NVG in high altitude area. But the former can effectively maintain visual acuity, and improve PEDF and VEGF levels in aqueous humor, with fewer complications and more accurate curative effect.
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@#AIM: To investigate the effect of transplanted pigmented epithelium-derived factor(PEDF)on human umbilical cord-derived mesenchymal stem cells(hUCMSCs)in the treatment of retinitis pigmentosa in rat models. <p>METHODS: The hUCMSCs were isolated and cultured, hUCMSCs were transfected with PEDF recombinant lentivirus. Experimental royal college of surgeon(RCS)rats were randomly divided into 3 groups, 8 rats in each group. The experimental group was injected with PEDF-hUCMSCs subcutaneously. The hUCMSCs control group was injected with the same amount of hUCMSCs. The PBS control group was injected with the same amount of PBS. At 4 and 8wk after injection, electroretinography(ERG), the thickness of retinal outer nuclear layer(ONL)and green fluorescent protein(GFP)staining were observed. Immunofluorescence staining of retinal sections and the phagocytosis of MERTK protein were evaluated to determine phagocytosis. <p>RESULTS: PEDF gene carrying recombinant lentiviral vector could efficiently infect hUCMSCs. After infection, hUCMSCs were sub cultured and the lentivirus prolonged the expression in the cells of the target gene. At 8wk after transplantation, the amplitude of b-wave in ERG were significantly higher in the experimental group than in the control groups. At 4 and 8wk after transplantation, morphological changes of ONL thickness in the experimental group were significantly higher than those in the control groups(all <i>P</i><0.05). At 4 and 8wk after transplantation, hUCMSCs infected by lentiviral vector carrying PEDF were found survival in the subretinal cavity by GFP staining, and the PEDF-hUCMSCs were found having phagocytosis for outer segment fragments of photorecepter cells by immunofluorescence staining. At 4 and 8wk after transplantation, the Protein band gray values in experimental group were significantly higher than that in control groups, which indicated that the expressions of MERTK protein in experimental group were also increased significantly, and the phagocytic capacity of RPE were improved. <p>CONCLUSION: Transplantation of pigment epithelium-derived protein modified human umbilical cord-derived mesenchymal stem cells has a protective effect on the retina of RCS rats.
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Objective To explore the mechanism of pigment epithelium-derived factor (PEDF) on human liver cells in mitochondrial apoptosis induced by arsenic. Methods Human normal liver cell line (HHL-5) was exposed to NaAsO2 for 24 h, the dose was 0 (control), 2, 5, 8 and 10 μmol/L, respectively. The mitochondrial membrane potential was determined by using flow cytometry, and mRNA levels of PEDF, Bcl-2 and Bax were assayed by Real-time fluorescent quantitative PCR. Results The mitochondrial membrane potential in 10 μmol/L arsenic group (562.33 ± 31.50) was decreased markedly in comparison with that of control group (688.67 ± 47.50, P<0.01), the levels in other groups (5 μmol/L: 634.50 ± 49.03, 8 μmol/L: 655.67 ± 13.65) were not significantly changed (P > 0.05). Compared with control group [(86.69 ± 12.19)％], the mRNA levels of PEDF decreased significantly (P<0.01) when the HHL-5 cells were cultivated with 5 μmol/L [(59.35 ± 11.21)％], 8μmol/L [(58.46 ± 8.69)％] and 10μmol/L [(51.57 ± 7.58)％] arsenic, respectively, and the PEDF mRNA level in 10 μmol/L group was the lowest; but there was no significant change in PEDF mRNA level [(81.52 ± 13.93)％] when HHL-5 cell was cultivated with 2μmol/L arsenic. Similarly, mRNA level of Bcl-2 in 10 μmol/L arsenic group [(53.53 ± 21 . 49 )％] was significantly decreased compared with that of the control group [(96.48 ± 31.14)％, P < 0.05]. Whereas, Bax mRNA levels in arsenic exposure groups [(65.14 ± 32.34)％, (71.81 ± 12.61)％, (72.22 ± 28.40)％] were not significantly changed in comparison with that of control group [(84.47 ± 35.38)％, P > 0.05]. Conclusion PEDF may play a role in mitochondrial apoptosis of human liver cells induced by arsenic through regulating Bcl-2 level.
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Pigment epithelium-derived factor(PEDF)is an endogenously produced protein widely expressed throughout the human body,which exhibits multiple and varied biological actions.Over the past few years,PEDF has been described as a multifaceted protein with anti-angiogenic, neurotrophic, neuroprotective, and anti-permeability.PEDF effectively inhibits vascular endothelial growth factor(VEGF)-driven angiogenesis and vascular permeability by regulating the intracellular proteolysis of VEGF receptors in re -cent reports.Moreover,PEDF can inhibit the occurrence of neural degenerative diseases through a variety of signaling pathways.The aim of the present review is to discuss the varied biological actions of PEDF and to identify the possible molecular mechanism underly -ing its effects.
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Neovascular eye diseases,with highly complicated and refractory,are one group of the most difficult problems for domestic and overseas clinical oculists in recent years.Angiogenesis is a complex process participating with multiple factors and various ways,and its exact pathogenesis is unclear yet.Ocular neovascularization is the dynamic balance result between proangiogenic factors and antiangiogenic factors provided that intraocular microenvironment be under a physiological condition.Some pathological factors including ischemia,hypoxia and inflammation destroy the dynamic balance between cytokines leading to neovascularization.Currently studies indicated that numerous cytokines containing vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) are closely linked to ocular neovascularization,while it is more crucial to elucidate the molecular mechanism of numerous cytokines involved in ocular neovascularization,which can recognize the pathogenesis of neovascular eye disease more clearly.Research progress of pathological relevant cytokines of neovascular eye diseases,including VEGF,insulin-like growth factor 1 (IGF-1),angiogenin-2 (Ang-2),stromal cell derived factor 1 (SDF-1) and PEDF were reviewed in this paper.
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ObJective To observe the inhibitory effects of hydrogen on corneal neovascularization (CNV) induced by alkali burn in rats and its underlying mechanisms Methods A total of 48 SPF-grade SD rats (48 eyes) were selected and randomly divided into blank group,control group and treatment group.Then the model of CNV was established successfully induced by alkali burn in these animals.The treatment group was given 1.6 × 10-6 hydrogen eye drops,the control group received tobramycin dexamethasone eye drops,and blank group left untreated.And CNV length was measured on day 1,3,7 and 14 after model establishment,followed by calculation of the CNV area.The corneal and aqueous humor of the rats was removed on day 14 after alkali burn.Then PEDF protein was detected by immunohistochemical staining,and RT-PCR was used to detect the expression of PEDF mRNA in the cornea.Additionally,TNF-α in the aqueous humor was detected by enzyme e-linked immunosorbent assay (ELISA).Resuits The area of corneal neovascularization on day 3,7 and 14 after alkali burn was (3.26 ± 1.82) mm2,(8.59 ± 3.98)mm2 and (3.24 ± 2.34) mm2 in the treatment group,respectively,(4.27 ± 2.68) mm2,(6.44 ± 4.67) mm2 and (15.98 ± 4.75) mm2 in the control group,respectively,(9.49 ±2.79)mm2,(17.71 ±4.06)mm2 and (25.35 ± 1.40)mm2 in blank group,respectively.The CNV areas of the treatment group and the control group were less than that of the blank group on day 3,7 and 14 after alkali bum,and the differences were statistically significant (all P ＜ 0.01),but there was no significant difference between the treatment group and the control group on day 3,7 after alkali bum (P ＞ 0.05),while the area of the treatment group was significantly less than that of the control group on 14th day,and the difference was statistically significant (P ＜0.01).On day 14 after alkali bum,the results of immunohistochemistry showed that the expression of PEDF protein in corneal epithelium was significantly increased in the treatment group than that in the control group and the blank group,and there were few new blood vessels in the stromal layer.RT-PCR results showed that the expression of PEDF mRNA in the treatment group was significantly higher than that in the control group and the blank group,and the difference was statistically significant (all P ＜ 0.01).ELISA results showed that the content of TNF-α protein in the treatment group was significantly lower than that in the control group and the blank group,and the difference was statistically significant (all P ＜0.01).Conclusion 1.6 × 10-6 hydrogen eye drops can effectively inhibit the corneal neovascularization which induced by alkali burn in rats.
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Objective To observe the clinical efficacy of Tangzhiping Prescription on non-proliferative diabetic retinopathy (NPDR) and the effects on serum vascular epithelial growth factor (VEGF) and pigment epithelium derived factor (PEDF). Methods Totally 86 NPDR patients were randomly divided into treatment group (43 cases) and control group (43 cases). Both groups were given hypoglycemic, anti hypertensive and lipid-regulatory basic therapy. The control group was given calcium distillate capsules, 0.5 g per time, 3 times a day, orally; the treatment group was treated with Tangzhiping Prescription based on the control group, 1 dosage per day, twice a day, orally. Four weeks were set as one treatment course. Treatment for both groups lasted for three courses. Clinical efficacy and fundus efficacy of both groups were evaluated. TCM symptom scores, fundus scores, and visual condition were observed; FPG, 2 h PG, Hb A1 C, TC, TG, HDL-C and LDL-C and changes in the contents of VEGF and PEDF were detected. Results The control group and the treatment group lost 2 and 3 cases respectively. The total effective rate of clinical efficacy and total fundus efficiency of the control group were 65.00％ and 68.35％, respectively, and the treatment group were 87.50％ and 84.62％ respectively, with statistical significance (P＜0.05). Compared with before treatment, TCM symptom scores and fundus scores decreased significantly after treatment in both groups (P＜0.01); After treatment, the TCM symptom scores and fundus scores in the treatment group were lower than those in the control group (P＜0.05, P＜0.01). Compared with before treatment, visual acuity improved significantly after treatment in both groups (P＜0.05, P＜0.01). Compared with before treatment, levels of FPG, 2 h PG, Hb A1 c, TC, TG, and LDL-C decreased significantly in both groups after treatment (P＜0.01). After treatment, the levels of 2 h PG, Hb A1 c, TC, TG, and LDL-C in the treatment group were significantly lower than the control group (P＜0.05, P＜0.01). Compared with before treatment, the levels of VEGF decreased and PEDF levels increased in both groups, with statistical significance (P＜0.01). After treatment, the improvement of VEGF and PEDF in the treatment group was better than that in the control group (P＜0.05, P＜0.01); negative correlation was found between VEGF and PEDF (r=-0.320, P＜0.01). Conclusion Tangzhiping Prescription can effectively improve the clinical symptoms of patients with NPDR and slow down the progress of NPDR via reducing the blood glucose and blood lipids, and regulating the contents of VEGF and PEDF.
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Objective To investigate the protective effects and possible mechanism of pigment epithelium derived faetor (PEDF) on myocardial cells H9C2 under hypoxia and serum-free condition.Methods H9C2 cells were culture in vitro and performed the hypoxia and serum-free processing.The cells were divided into the control group (H9C2),hypoxia group (hypoxia + H9C2),PEDF group(hypoxia+H9C2 +PEDF) and mitochondrial fission inhibitor(Mdivi-1) group(hypoxia+h9C2+Mivi-1).The apoptotic rate was detected by TUNNEL staining.The proteins levels of dynamin related peptide1 (Drp1) and cleaved-caspase 3 were measured by Western blot.Electron Microscopy and MitoTracker Red were used to detect the mitochondria morphology,the mitochondrial membrane potential was evaluated by cationic dye JC-1.MitoSOXTM was used to detect mitochondrial reactive oxygen species (ROS).Results Hypoxia induced mitochondrial fission(P＜0.05).The hypoxia group (6 h) and control group had statistical difference(P＜0.05).PEDF reduces mitochondrial fission under hypoxia condition(P＜0.05),which had statistical difference between the PEDF group and hypoxia group (6 h)(P＜0.05).PEDF and Mdivi-1 could decrease cell apoptosis under hypoxia condition(24 h),compared with the hypoxia group,the difference was statistically significant(P＜0.05).Conclusion PEDF decrease cell apoptosis by inhibiting H9C2 cells mitochondrial fission under hypoxia condition.
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@#AIM:To observe the protective effect of pigment epithelium derived factor(PEDF)on retinal ganglion cells(RGCs)after acute injury of optic nerve in rats. <p>METHODS: Totally 60 healthy female SD rats were randomly divided into three groups: blank group, model group and experimental group, 20 rats in each group. Acute injury model of optic nerve in rats are established by crushing optic nerve. After the model of optic nerve injury was made successfully in model group, the rats were injected with balanced salt solution 5μL into vitreous cavity, and at 1 and 2wk later respectively they were injected again. In experimental group, after the model of optic nerve injury was made successfully, immediately the rats were injected with the PEDF 5μL into vitreous cavity, also at 1 and 2wk later respectively they were injected again. To observe the retinal morphologic changes and count the number of retinal ganglion cells, the specimen was given HE staining and observed under light microscope. The immunohistochemical semi quantitative method was used to detect the expression of Bcl-2 and Bax to observe the effect of PEDF on the optic nerve after injury. <p>RESULTS: Cell morphology of ganglion cells in experimental group was better and the number of RGCs was much more, comparing with the model group by HE staining. The expression of Bcl-2 and Bax in the experimental group was significantly different with that in the blank group and the model group(<i>P</i><0.05). The expression of Bcl-2 increased and the expression of Bax decreases in the experimental group compared with the model group. <p>CONCLUSION: PEDF can inhibit or weak the apoptosis of RGCs cells after optic nerve injury and alleviate the injury of optic nerve by increasing the expression of Bcl-2 protein or decreasing the expression of Bax protein.
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Objective To investigate the protective effects and possible mechanism of pigment epithelium derived faetor (PEDF) on myocardial cells H9C2 under hypoxia and serum-free condition.Methods H9C2 cells were culture in vitro and performed the hypoxia and serum-free processing.The cells were divided into the control group (H9C2),hypoxia group (hypoxia + H9C2),PEDF group(hypoxia+H9C2 +PEDF) and mitochondrial fission inhibitor(Mdivi-1) group(hypoxia+h9C2+Mivi-1).The apoptotic rate was detected by TUNNEL staining.The proteins levels of dynamin related peptide1 (Drp1) and cleaved-caspase 3 were measured by Western blot.Electron Microscopy and MitoTracker Red were used to detect the mitochondria morphology,the mitochondrial membrane potential was evaluated by cationic dye JC-1.MitoSOXTM was used to detect mitochondrial reactive oxygen species (ROS).Results Hypoxia induced mitochondrial fission(P＜0.05).The hypoxia group (6 h) and control group had statistical difference(P＜0.05).PEDF reduces mitochondrial fission under hypoxia condition(P＜0.05),which had statistical difference between the PEDF group and hypoxia group (6 h)(P＜0.05).PEDF and Mdivi-1 could decrease cell apoptosis under hypoxia condition(24 h),compared with the hypoxia group,the difference was statistically significant(P＜0.05).Conclusion PEDF decrease cell apoptosis by inhibiting H9C2 cells mitochondrial fission under hypoxia condition.