Role and mechanism of platelet-derived growth factor BB in thrombocytosis in Kawasaki disease / 中国当代儿科杂志

OBJECTIVES@#To study the role and mechanism of platelet-derived growth factor BB (PDGF-BB) on platelet production in Kawasaki disease (KD) mice and human megakaryocytic Dami cells through in vitro and invivo experiments.@*METHODS@#ELISA was used to measure the expression of PDGF in the serum of 40 children with KD and 40 healthy children. C57BL/6 mice were used to establish a model of KD and were then randomly divided into a normal group, a KD group, and an imatinib group (30 mice in each group). Routine blood test was performed for each group, and the expression of PDGF-BB, megakaryocyte colony forming unit (CFU-MK), and the megakaryocyte marker CD41 were measured. CCK-8, flow cytometry, quantitative real-time PCR, and Western blot were used to analyze the role and mechanism of PDGF-BB in platelet production in Dami cells.@*RESULTS@#PDGF-BB was highly expressed in the serum of KD children (P<0.001). The KD group had a higher expression level of PDGF-BB in serum (P<0.05) and significant increases in the expression of CFU-MK and CD41 (P<0.001), and the imatinib group had significant reductions in the expression of CFU-MK and CD41 (P<0.001). In vitro experiments showed that PDGF-BB promoted Dami cell proliferation, platelet production, mRNA expression of PDGFR-β, and protein expression of p-Akt (P<0.05). Compared with the PDGF-BB group, the combination group (PDGF-BB 25 ng/mL + imatinib 20 μmol/L) had significantly lower levels of platelet production, mRNA expression of PDGFR-β, and protein expression of p-Akt (P<0.05).@*CONCLUSIONS@#PDGF-BB may promote megakaryocyte proliferation, differentiation, and platelet production by binding to PDGFR-β and activating the PI3K/Akt pathway, and the PDGFR-β inhibitor imatinib can reduce platelet production, which provides a new strategy for the treatment of thrombocytosis in KD.

Effect of platelet-derived growth factor-BB on pulmonary vascular remodeling in neonatal rats with hypoxic pulmonary hypertension and its mechanism / 中国当代儿科杂志

OBJECTIVES@#To study the effect of platelet-derived growth factor-BB (PDGF-BB) on pulmonary vascular remodeling in neonatal rats with hypoxic pulmonary hypertension (HPH).@*METHODS@#A total of 128 neonatal rats were randomly divided into four groups: PDGF-BB+HPH, HPH, PDGF-BB+normal oxygen, and normal oxygen (n=32 each). The rats in the PDGF-BB+HPH and PDGF-BB+normal oxygen groups were given an injection of 13 μL 6×1010 PFU/mL adenovirus with PDGF-BB genevia the caudal vein. After 24 hours of adenovirus transfection, the rats in the HPH and PDGF-BB+HPH groups were used to establish a neonatal rat model of HPH. Right ventricular systolic pressure (RVSP) was measured on days 3, 7, 14, and 21 of hypoxia. Hematoxylin-eosin staining was used to observe pulmonary vascular morphological changes under an optical microscope, and vascular remodeling parameters (MA% and MT%) were also measured. Immunohistochemistry was used to measure the expression levels of PDGF-BB and proliferating cell nuclear antigen (PCNA) in lung tissue.@*RESULTS@#The rats in the PDGF-BB+HPH and HPH groups had a significantly higher RVSP than those of the same age in the normal oxygen group at each time point (P<0.05). The rats in the PDGF-BB+HPH group showed vascular remodeling on day 3 of hypoxia, while those in the HPH showed vascular remodeling on day 7 of hypoxia. On day 3 of hypoxia, the PDGF-BB+HPH group had significantly higher MA% and MT% than the HPH, PDGF-BB+normal oxygen, and normal oxygen groups (P<0.05). On days 7, 14, and 21 of hypoxia, the PDGF-BB+HPH and HPH groups had significantly higher MA% and MT% than the PDGF-BB+normal oxygen and normal oxygen groups (P<0.05). The PDGF-BB+HPH and HPH groups had significantly higher expression levels of PDGF-BB and PCNA than the normal oxygen group at all time points (P<0.05). On days 3, 7, and 14 of hypoxia, the PDGF-BB+HPH group had significantly higher expression levels of PDGF-BB and PCNA than the HPH group (P<0.05), while the PDGF-BB+normal oxygen group had significantly higher expression levels of PDGF-BB and PCNA than the normal oxygen group (P<0.05).@*CONCLUSIONS@#Exogenous administration of PDGF-BB in neonatal rats with HPH may upregulate the expression of PCNA, promote pulmonary vascular remodeling, and increase pulmonary artery pressure.

Shexiang Baoxin Pill Regulates Intimal Hyperplasia, Migration, and Apoptosis after Platelet-Derived Growth Factor-BB-Stimulation of Vascular Smooth Muscle Cells via miR-451 / 中国结合医学杂志

OBJECTIVE@#To investigate the regulatory roles of Shexiang Baoxin Pill (SXBXW) in neointimal formation and vascular smooth muscle cells (VSMCs) invasion and apoptosis as well as the potential molecular mechanisms using cultured VSMCs model of vascular injury (platelet-derived growth factor (PDGF)-BB-stimulated) in vitro.@*METHODS@#VSMCs were randomly assigned to 5 groups: blank, PDGF-BB (20 ng/mL+ 0.1% DMSO), SXBXW-L (PDGF-BB 20 ng/mL + SXBXW low dose 0.625 g/L), SXBXW-M (PDGF-BB 20 ng/mL + SXBXW medium dose 1.25 g/L) and SXBXW-H (PDGF-BB 20 ng/mL+ SXBXW high dose 2.5 g/L) group. Cell proliferation was assessed using cell counting kit-8 (CCK-8) assay and bromodeoxyuridine (BrdU) incorporation assay, the migration effects were detected by Transwell assay, cell apoptosis rate was measured by the Annexin V/propidium iodide (PI) apoptosis kit. The markers of contractile phenotype of VSMCs were detected with immunofluorescent staining. To validate the effects of miR-451 in regulating proliferation, migration and apoptosis treated with SXBXW, miR-451 overexpression experiments were performed, the VSMCs were exposed to PDGF-BB 20 ng/mL + 0.1% DMSO and later divided into 4 groups: mimic-NC (multiplicity of infection, MOI=50), SXBXW (1.25 g/L) + mimic-NC, mimic-miR451 (MOI=50), and SXBXW (1.25 g/L) + mimic-miR451, and alterations of proteins related to the miR-451 pathway were analyzed using Western blot.@*RESULTS@#PDGF-BB induced VSMCs injury causes acceleration of proliferation and migration. SXBXW inhibited phenotypic switching, proliferation and migration and promoted cell apoptosis in PDGF-BB-induced VSMCs. In addition, miR-451 was shown to be down-regulated in the VSMCs following PDGF-BB stimulation. SXBXW treatment enhanced the expression of miR-451 in PDGF-BB-induced VSMCs (P<0.05). Compared with SXBXW + mimic-NC and mimic-miR451 groups, the expression of tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (Ywhaz) and p53 was further reduced in SXBXW + mimic-miR451 group, while activating transcription factor 2 (ATF2) was increased in VSMCs (P<0.05).@*CONCLUSION@#SXBXW regulated proliferation, migration and apoptosis via activation of miR-451 through ATF2, p53 and Ywhaz in PDGF-BB-stimulated VSMCs.

Study of rrPDGF-BB gene modified BMSCs in promoting distraction osteogenesis of rat femur / 中华显微外科杂志

Objective:To investigate the therapeutic effect of rat bone marrow mesenchymal stem cells (BMSCs) transfected with recombinant rat platelet-derived growth factor BB (rrPDGF-BB) gene on the distraction osteogenesis.Methods:From October, 2019 to June, 2020, 48 batches of BMSCs were cultured from 48 young SD rats, 24 of which were transfected with rrPDGF-BB gene by lentivirus. Meanwhile, other 72 male adult SD rats were randomly selected to establish the right femoral distraction osteogenesis model. The rats were equally divided into 3 groups. PBS, BMSCs without intervention and BMSCs transfected with rrPDGF-BB gene were injected into the distraction space of each group of rats assigned as Blank group, Negative group and Experimental group, respectively. Results of the experiment were evaluated by means of imaging and immunohistochemistry. P<0.05 indicated a statistically significant difference. Results:The cultured BMSCs grew well. The expression of CD34(0.1%) and CD45(2.8%) in the third generation of BMSCs was low, and that of CD29 (95.1%) was high, which was consistent with the phenotype of BMSCs described in literatures. After transfection, the expression of green fluorescence gradually increased with the extension of transfection time, confirming the success of transfection. After 14 days, all rats reached the expected distance of distraction. The rats were observed at assigned time points in 2, 4 and 8 weeks. The photos of femur specimen showed that continuous callus could be seen in the experimental group, the hardness and colour were close to the normal bone tissue, and the activity of the distraction space was poor, which was lower than that of the blank group. X-ray examination showed that there were more new callus in the experimental group, and the bone marrow cavity was re-canalized earlier than that of the blank group; Micro-CT examination, in sagittal plane, showed that the distraction space of the experimental group healed well, the broken end was connected, and the recanalization of bone marrow cavity was earlier than that of the blank group; Micro-CT parameters of each group showed that trabecular thickness[(0.297±0.005) mm], trabecular number [(1.663±0.032) mm], bone volume fraction[(59.832±2.187)%] and bone mineral density[(0.586±0.014) g/cm 3] of the experimental group were the greatest, while trabecular separation[(0.399±0.051) mm] of the experimental group was the smallest. There was statistical difference between each group( P < 0.05); HE staining and VEGF immunohistochemistry showed that the vessels and chondrocytes formed earlier and were more in the experimental group than that in the blank group. In 8 weeks, the new callus joined into one piece under the microscope in the experimental group, and the bone marrow cavity was re-canalized with a large number of red blood cells. Conclusion:Studies have shown that BMSCs transfected with rrPDGF-BB gene can promote the formation of callus in the distraction area of rats, shorten the mineralisation time of new callus, and promote the maturation of new bone in the area of distraction osteogenesis.

Effects of hypoxia inducible factor-1α on osteogenic differentiation and angiogenesis related factors of bone marrow mesenchymal stem cells / 口腔疾病防治

Objective@#To investigate the level of hypoxia inducible factor-1α (HIF-1α) on osteoblasts and angiogenesis-associated cytokines in bone marrow mesenchymal stem cells (BMSCs) from SD rats.@*Methods@#BMSCs were isolated and cultured and identified by flow cytometry. Plasmid vectors containing upregulated and downregulated HIF-1α gene and a control vector were constructed. The plasmids were transfected into BMSCs by Lipofectamine®LTX transfection reagent, and the cells were divided into an overexpression experimental group, an overexpression control group, a low expression experimental group and a low expression control group. All components were stained with a lizarin red 3 d and 7 d after osteogenesis induction. The mRNA expression levels of the target gene HIF-1α, osteogenic differentiation-specific markers, including Runt-related transcription factor 2 (Runx2) and angiogenic markers, including platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-β (TGF-β), were detected by RT-PCR. Western blot was used to detect the protein expression of the target proteins HIF-1α, Runx2, and PDGF-BB.@*Results@# The CD29- and CD45-positivity rates of BMSC surface markers identified by flow cytometry were 98.2% and 4.2%, respectively. RT-PCR results showed that the mRNA expression of HIF-1α, Runx2, TGF-β and PDGF-BB was observably increased (P < 0.001). The mRNA expression levels of HIF-1α, Runx2, TGF-β and PDGF-BB in BMSCs from the low expression experimental group were significantly reduced (P < 0.001). Western blot results showed that the expression levels of HIF-1α, Runx2 and PDGF-BB in BMSCs from the overexpression experimental group were all increased (P < 0.001). The expression levels of HIF-1α, Runx2 and PDGF-BB in BMSCs from the low expression experimental group were reduced (P < 0.001). Alizarin red staining results showed that the area of calcium nodules in the low expression experimental group was smaller than that in low expression control group, the area of red calcium nodules in the over expression experimental group was larger than that in over expression control group, and with the increase of osteogenic induction time, the calcification area of each group also increased.@*Conclusion@# Upregulation and downregulation of HIF-1α can regulate the osteogenic differentiation and the expression of angiogenesis related factors of BMSCs.

Effects of vascular endothelial growth factor combined with platelet-derived growth factor-bb on the angiogenic and proliferative abilities of bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND: Bone tissue engineering provides a new path to treat critical-sized bone defects. However, a stable osteogenesis and osseointegration can only be insured via formation of an intact vascular network, which can achieve satisfactory therapeutic effects. Thereafter, angiogenesis is a challenge and difficulty in bone tissue engineering. OBJECTIVE: To investigate the effect of combined application of vascular endothelial growth factor (VEGF) and platelet-derived growth factor-BB (PDGF-BB) on proliferation and angiogenesis of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells from Sprague-Dawley rats were cultured and isolated in vitro, and interfered with different concentrations of VEGF (20, 40, 60, 80, 100, 120 μg/L) and PDGF-BB (20, 40, 60, 80, 100, 120 μg/L) in combination or 100 μg/L VEGF, and 100 μg/L PDGF-BB alone. The optimum concentration of promoting cell proliferation was detected by cell counting kit-8 assay. The expression levels of angiopoietin-1, hypoxia-inducible factor-1α, hepatocyte growth factor and insulin-like growth factor were detected by RT-PCR at 7 and 14 days after intervention. RESULTS AND CONCLUSION: (1) After growth factors were added, the cell proliferative ability was significantly improved, and combined application revealed better effect. The optimal concentration grou was 80 μg/L VEGF+80 μg/L PDGF-BB. (2) Both VEGF and PDGF-BB could promote the mRNA expression levels of angiopoietin-1, hypoxia-inducible factor-1α, hepatocyte growth factor and insulin-like growth factor, and the effect was more obvious in combined application. (3) The mRNA expression levels of hypoxia-inducible factor-1α and hepatocyte growth factor were significantly increased with time (P < 0.05). The mRNA expression levels of angiopoietin-1 and insulin-like growth factor were significantly decreased with time (P < 0.05). (4) In vitro experimental results suggest that VEGF and PDGF-BB at the concentration of 80 μg/L can consistently promote angiogenesis, and the effect of combined application is better than that of single application.

Platelet-derived growth factor-bb promotes proliferation, differentiation and migration of skeletal muscle myoblast / 中国组织工程研究

BACKGROUND: Skeletal muscle myoblasts differentiate and fuse to form polynuclear muscle tubes and muscle fibers to complete the repair of muscle injury when skeletal muscles were injured. However, the repair process is slow and incomplete. Platelet-derived growth factor-BB can stimulate the proliferation, differentiation and migration of a variety of tissue cells, and plays an important role in the process of tissue repair after various injuries. OBJECTIVE: To explore the effects of different concentrations of platelet-derived growth factor-BB on proliferation, differentiation and migration of skeletal muscle myoblast C2C12 cells and the action mechanism. METHODS: The mouse skeletal muscle myoblast C2C12 cells were cultured with platelet-derived growth factor-BB 0, 5, 10, 20 and 40 μg/L and platelet-derived growth factor receptor inhibitor imatinib. The expression of platelet-derived growth factor receptor in C2C12 cells was detected by immunocytochemistry and western blot assay. After 1, 2, 3, 4, and 5 days of culture, CCK8 method was used to detect cell proliferation. After 4 days of induction and differentiation culture, the formation of myotubes in each group was observed by light microscope. The expression of myosin heavy chain and MyoG Gene was observed by immunofluorescence and western blot assay. After 48 hours of culture, Transwell method was used to detect cell migration. RESULTS AND CONCLUSION: (1) Immunofluorescence chemistry and western blot assay indicated that platelet-derived growth factor receptor expression could be detected in C2C12 cells, and semi-quantitative statistical analysis showed no significant difference in platelet-derived growth factor receptor expression between groups with different platelet-derived growth factor-BB concentrations (P > 0.05). (2) CCK8 assay demonstrated that the proliferation of C2C12 cells showed no significant change in platelet-derived growth factor-BB groups compared with the control group (platelet-derived growth factor-BB 0 μg/L group). (3) Immunofluorescence chemistry showed that compared with control group, number of myosin heavy chain positive cells increased in platelet-derived growth factor-BB groups; 40 μg/L platelet-derived growth factor-BB concentration got the highest; the mature muscle tubes up to (27.00±0.76) per field of vision, and MyoG expression populations was most obviously compared with the control group (P < 0.05). (4) Transwell results showed that compared with the control group, the migration number of C2C12 cells in the platelet-derived growth factor-BB group increased, and the migration number in the 40 μg/L group was up to 144.00±13.03 (P < 0.05). (5) It is concluded that platelet-derived growth factor-BB promoted the migration, differentiation and myotube formation of C2C12 cells, and the pro-differentiation mechanism is related to its enhanced binding with platelet-derived growth factor receptor, so as to improve the expression of differentiation related gene MyoG.

Effect of Korean Magnolia obovata Extract on Platelet-Derived Growth Factor-Induced Vascular Smooth Muscle Cells / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of Korean Magnolia obovata crude extract (KME) on plateletderived growth factor (PDGF)-BB-induced proliferation and migration of vascular smooth muscle cells (VSMCs).@*METHODS@#KME composition was analyzed by high-performance liquid chromatography (HPLC). VSMCs were isolated from the aorta of a Sprague-Dawley rat, incubated in serum free-Dulbecco's modified Eagle's medium in the presence or absence of KME (10, 30, 100, and 300 μg/mL), then further treated with PDGF-BB (10 ng/mL). VSMC proliferation was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and VSMC migration was determined using the Boyden chamber and scratch wound healing assays. Western blot analysis was used to detect phosphorylation of extracellular signal-regulated protein kinases 1 and 2 (p-ERK1/2), protein kinase B (p-Akt), and stress-activated protein kinase/c-Jun NH2-terminal kinase (p-SAPK/JNK). The antimigration and proliferation effects of KME were tested using aortic sprout outgrowth.@*RESULTS@#The HPLC analysis identified honokiol (0.45 mg/g) and magnolol (0.34 mg/g) as the major components of KME. KME (30, 100, and 300 μg/mL) significantly decreased the proliferation and migration of PDGF-BB-stimulated (10 ng/mL) VSMCs and the PDGF-BB-induced phosphorylation of EKR1/2, Akt, and SAPK/JNK (P<0.05). Furthermore, PDGF-BBinduced VSMCs treated with 300 μg/mL of KME showed reduction in aortic sprout outgrowth.@*CONCLUSION@#KME could inhibit abnormal proliferation and migration of VSMCs by down-regulating the phosphorylation of EKR1/2 and Akt. Thus, KME might be a functional food for preventing vascular disorders.

Study on serum PDGF-BB as biomarker for diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To observe the correlation between serum level of platelet derived growth factor -BB(PDGF-BB)and diabetic retinopathy(DR)in type 2 diabetic patients.<p>METHODS: Seventy-five cases of healthy subjects underwent physical examination were taken as control group. Diabetic patients were divided into non-diabetic retinopathy group(NDR, 25 cases), non-proliferative diabetic retinopathy group(NPDR, 25 cases)and proliferative diabetic retinopathy group(PDR, 25 cases). Serum level of each group was measured using Enzyme-Linked Immunosorbent Assay(ELISA). Correlation between serum PDGF-BB level and stages of diabetes mellitus, macular thickness, glycosylated hemoglobin, and other biochemical indexes in serum were analyzed. <p>RESULTS: Serum level of PDGF-BB in control group(400.28±44.55pg/mL), NDR(409.65±50.37pg/mL), NPDR(535.67±69.21pg/mL)and PDR subgroup(551.60±103.46pg/mL)were significantly different(<i>F</i>=14.259, <i>P</i><0.01). Serum level of PDGF-BB in NPDR group and PDR group were higher compared to control group and NDR subgroup. Multiple regression analysis showed that serum level of PDGF-BB positively correlated with fasting blood glucose(FBG), triglycerides(TG)and stages of diabetes(<i>P</i><0.05). Serum level of PDGF-BB in PDR subgroup correlated with the thickness of macular(<i>r=</i>0.613, <i>P</i><0.05). Whereas, this correlation was not observed in other groups(<i>r_C=</i>0.013, <i>r_NDR=</i>0.051,<i> r_NPDR=</i>0.062; <i> P</i>>0.05).<p>CONCLUSION: Serum PDGF-BB level was observed to rise with severity of DR and relevant to macula edema. It was also positively correlated to FPG, TG and stages of diabetes. PDGF-BB could be recommended as diagnostic biomarker for DR.

Detection and significance of LCN2 and PDGF-BB in serum and metastasis of lung cancer patients with bone metastasis / 临床与实验病理学杂志

Purpose To investigate the expression of lipocalin-2 (LCN2) and plateled derived growth factor-BB (PDGF-BB) in serum,carcinoma and bone metastases of lung cancer patients. Methods Protein chip were used to screen the differential expression of cytokines in serum of 19 lung cancer patients (9 patients with bone metastasis and 10 patients freedistant metastasis) . Immunohistochemistry was performed to assess the differential expression of LCN2 and PDGF-BB cytokines in 12 cases of primary lung cancer without distant metastasis and 12 cases of primary lung cancer with only bone metastasis. Results Serum level of lipid transport factor (LCN2) and PDGFBB in non-small cell lung cancer patients with bone metastasis were significantly higher than that without distant metastasis(P< 0. 05) . There was no difference cytokines between small cell lung cancer patients with bone metastasis and without metastasis group (P > 0. 05) . The results of immunohistochemistry showed that high expression of LCN2 and PDGF-BB in bone metastasis tissues was significantly higher than that in primary lung cancer tissues. Conclusions High expression of LCN2 and PDGF-BB in serum and bone metastasis tissue of patients with non-small cell lung cancer might be involved in the occurrence,development of bone metastasis of lung cancer in the bone marrow,may be an important biomarker and potential therapeutic target for bone metastasis of lung cancer.

Predicting value of PDGF-BB combined with CLIF-COFs for the occurrence of ARDS in post-liver transplantation patients / 国际外科学杂志

Objective To investigate the role of cytokines combined with CLIF consortium organ failure score (CLIF-COFs) for predicting the occurrence of acute respiratory distress syndrome (ARDS) in for post-liver transplant for hepatitis B-related acute-on-chronic liver failure (HB-ACLF) patients.Methods From Jul.2014 to Oct.2017,there were 37 cases of HB-ACLF undergoing liver transplantation in Beijing YouAn Hospital,Capital Medical University.According to whether the patients happened ARDS or not,37 cases were divided into ARDS group (n =9) and non-ARDS group (n =28).All patients' plasma was prospectively collected immediately before liver transplantation and on the I st,3rd,5th,7th day post-liver transplantation.The serum levels of twenty-seven cytokines were determined by 200 LUMINEX liquid chip technology.Cytokines,CLIF-COFs,clinical and biochemical indexes were analyzed with logistic regression and the receiver operating characteristic (ROC) to confirm the correlation with ARDS post liver transplantation.Results There were significant differences between HB-ACLF patients between ARDS group and non-ARDS group in age and pre-transplant infection (P ＜ 0.05).The CLIF-COFs was higher in ARDS non-without than that in non-ARDS group (P =0.019).The serum levels of vascular endothelial growth factor and platelet-derived growth factor bb were lower but IL-6 was higher post transplantation in ARDS group.The COX analysis indicated that CLIF-COFs and post liver transplantation PDGF-BB were predictors of post-LT ARDS.The area under the receiver operating characteristic (AUROC) curves was 0.728 and 0.175,respectively.The area under the curve of the discriminatory power of CLIF-COFs combined with PDGF-BB was 0.913,and the maximum Youden index is 0.786.Conclusion CLIF-COFs combines with PDGF-BB can predict the occurrence of ARDS post-liver transplantation in HB-ACLF patients.

The effects of macrophages with high expression of TL1A on activation and proliferation of hepatic stellate cells in vitro / 中华肝脏病杂志

Objective@#To explore the effects of macrophages with high expression of TL1A on the activation and proliferation of HSCs in vitro. @*Methods@#The Bone marrow-derived macrophages (BMMs) and peritoneal macrophages (PMs) from wild type (WT) and myeloid-overexpressed TL1A transgenic mice were isolated, differentiated and activated. HSCs were harvested from activated macrophages culture supernatant (CM). HSCs were detected by immunofluorescence and real-time Q-PCR. And the proliferation was detected by CCK-8 and BrdU assay kit. The levels of IL-1β and PDGF-BB in macrophage culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA). @*Results@#BMMs-derived CM-intervention HSCs were used to detect the expression of α-smooth muscle actin (α-SMA) on the 2nd, 4th and 6th day respectively by immunofluorescence method. There was no significant difference between the two groups on the 2 nd and the 6th day, P > 0.05; On day 4, the CM/Tg group was significantly higher than that of CM/WT group, P < 0.01; the results of CMs derived from PMs were consistent with the above trend. The expression of α-SMA mRNA on the 2nd, 4th and 6th day was detected by real-time Q-PCR method using BM-derived CMs. No significant difference was found between the groups on the 2nd day (P > 0.05).α-SMA mRNA increased further on the 4th and 6th day, and the level of CM/Tg in CM/Tg group was significantly higher than that in CM/WT group (P < 0.05). The detection results of CMs derived from PMs were consistent with the above trend. The results of CCK-8 assay and BrdU assay showed that the proliferation rate of HSCs in CM Tg group was significantly higher than that in CM/WT group (P < 0.01). The CMs derived from PMs were used to interfere with HSCs. And the results were consistent with the above trend. For BMMs, the levels of IL-1β and PDGF-BB in the lipopolysaccharide (LPS) + IFNγ/Tg culture supernatant were significantly higher than those in the LPS+IFNγ/WT group (P < 0.01). For the culture supernatants of PMs Liquid test results consistent with the above trend. @*Conclusion@#Macrophages with high expression of TL1A could enhance the activation and proliferation of HSCs by increasing the secretion of IL-1β and PDGF-BB.

Laminin and Platelet-Derived Growth Factor-BB Promote Neuronal Differentiation of Human Urine-Derived Stem Cells

Urine-derived stem cells (USCs) are considered as a promising cell source capable of neuronal differentiation. In addition, specific growth factors and extracellular matrix are essential for enhancing their neuronal differentiation efficiency. In this study, we investigated the possibility of neuronal differentiation of USCs and the role of laminin and platelet-derived growth factor BB (PDGF-BB) as promoting factors. USCs were isolated from fresh urine of healthy donors. Cultured USCs were adherent to the plate and their morphology was similar to the cobblestone. In addition, they showed chromosome stability, rapid proliferation rate, colony forming capacity, and mesenchymal stem cell characteristics. For inducing the neuronal differentiation, USCs were cultured for 14 days in neuronal differentiation media supplemented with/without laminin and/or PDGF-BB. To identify the expression of neuronal markers, RT-PCR, flow cytometry analysis and immunocytochemistry were used. After neuronal induction, the cells showed neuron-like morphological change and high expression level of neuronal markers. In addition, laminin and PDGF-BB respectively promoted the neuronal differentiation of USCs and the combination of laminin and PDGF-BB showed a synergistic effect for the neuronal differentiation of USCs. In conclusion, USCs are noteworthy cell source in the field of neuronal regeneration and laminin and PDGF-BB promote their neuronal differentiation efficiency.

The relationship between plasma PDGF-BB level and coronary heart disease and coronary artery stenosis / 天津医药

Objective To investigate the relationship between plasma level of platelet derived growth factor-BB (PDGF-BB), coronary heart disease (CHD) and the severity of coronary artery stenosis. Methods A total of 262 patients hospitalized in Department of Cardiology, Tianjin Chest Hospital were collected in this study. According to the medical history, symptoms, laboratory examination and the results of coronary angiography, patients were divided into stable angina pectoris (SAP) group (n=57), acute coronary syndrome (ACS) group (n=119) and normal control group (n=86). The ACS group was divided into three subgroups:single vessel group (n=38), double vessel group (n=35) and multiple vessel group (n=46). The general clinical data, biochemical parameters and plasma PDGF-BB levels were compared between SAP group, ACS group and control group. Spearman correlation analysis was used to analyze the relationship between PDGF-BB level, high-sensitivity C-reactive protein (hs-CRP) and Gensini scores. Logistic regression analysis was used to analyze the risk factors of coronary heart disease. Results (1) The plasma levels of hs-CRP and PDGF-BB were significantly higher in ACS group than those in control group and SAP group (P0.05). (3) There was no significant difference in plasma level of PDGF-BB between single vessel group, double vessel group and multiple vessel group (P > 0.05). (4) Logistic regression analysis showed that high plasma level of PDGF-BB was the risk factor for coronary heart disease. Conclusion PDGF-BB plasma level is associated with the pathogenesis of coronary heart disease, which may reflect the instability of coronary atherosclerotic plaques, but it is not an index to evaluate the severity of coronary stenosis.

Platelet-Derived Growth Factor-BB-Immobilized Asymmetrically Porous Membrane for Enhanced Rotator Cuff Tendon Healing

Rotator cuff tear is a common musculoskeletal disease that often requires surgical repair. Despite of recent advances in surgical techniques, the re-tear rate of the rotator cuff tendon is very high. In this study, a platelet-derived growth factor-BB (PDGF-BB)-immobilized asymmetrically porous membrane was fabricated to investigate the feasibility for enhancing rotator cuff tendon regeneration through the membrane. PDGF-BB is recognized to promote tendon regeneration. The asymmetrically porous membrane was fabricated by polycaprolactone and Pluronic F127 using an immersion precipitation technique, which can allow selective permeability (preventing scar tissue invasion into defect region but allowing permeation of oxygen/nutrients) and incorporation of bioactive molecules (e.g., PDGF-BB) via heparin binding. The PDGF-BB immobilized on the membrane was released in a sustained manner over 42 days. In an animal study using Sprague-Dawley rats, the PDGF-BB-immobilized membrane group showed significantly greater regeneration of rotator cuff tendon in histological and biomechanical analyses compared with the groups of control (suturing) and membrane without PDGF-BB immobilization. The enhancing regeneration of rotator cuff tendon of the PDGF-BB-immobilized membrane may be caused from the synergistic effect of the asymmetrically porous membrane with unique properties (selective permeability and hydrophilicity) as a scaffold for guided tendon regeneration and PDGF-BB sustainedly released from the membrane.

Optimization of Recombinant Human Platelet-Derived Growth Factor-BB Encapsulated in Poly (Lactic-co-Glycolic Acid) Microspheres for Applications in Wound Healing

Growth factors play multiple and critical roles in wound repair processes. Platelet-derived growth factor (PDGF) is a potent growth factor that is particularly important in the early inflammatory phase of wound healing. In order to extend the half-life of PDGF, polymeric encapsulation is used. In the current study, Poly (lactic-co-glycolic acid) (PLGA) microspheres containing recombinant human (rh) PDGF-BB were prepared to prolong the effectiveness of this growth factor. PLGA microspheres were optimized using a modified w/o/w-double-emulsion/solvent evaporation method by changing the processing conditions of stirring speed and emulsifier (polyvinyl alcohol) concentration. Microspheres prepared using the optimized method released rhPDGF-BB for up to three weeks. An in vitro migration assay showed a significant decrease in the wound area in cells treated with rhPDGF-BB microspheres compared to control cells. These findings demonstrate the potential of rhPDGF-BB encapsulated in microspheres to enhance wound healing.

The anti-fibrosis effect of ghrelin in human hepatic stellate cells via PI3K-AKT pathway / 中国综合临床

Objective To observe the effect of ghrelin on the expression of procollagen type Ⅰ and alpha smooth muscle actin (α-SMA) synthesis in vitro cultured human hepatic stellate cell (HSC-LX2) stimulated by Platelet-derived growth factor-BB (PDGF-BB).Besides,the effect of PI3K-AKT pathway was studied.Methods Cultured LX2 were intervented and jointing intervented according to the different ghrelin concentration by ghrelin and PDGF:control group,0.1 μmol/L Ghrelin group,10 μg/L PDGF group,0.05 μmol/L Ghrelin +10 μg/L PDGF group,0.1 μmol/L Ghrelin + 10 μg/L PDGF group,0.15 μmol/L Ghrelin + 10 μg/L PDGF group.Culture HSC-LX2 in vitro,joint intervention cells with different concentrations.Procollagen Ⅰ mRNA expression were detected by Polymerase chain reaction (PCR),besides,α-SMA and AKT expression were detected by Western blot in each groups.After treatment by PI3K specific inhibitor LY294002 in LX2,three groups were divided into PDGF,Ghrelin + PDGF and LY294002 + Ghrelin +PDGF.Procollagen Ⅰ mRNA expression were detected by PCR,and α-SMA was detected by Western blot.Results PCR results showed that procollagen Ⅰ expression in PDGF treated group was significantly higher than the control group ((6.91 ± 0.46) vs.(1.00 ± 0.08),P ＜ 0.05),so PDGF can promote the expression of procollagen type Ⅰ.Procollagen Ⅰ mRNA expression between ghrelin group(0.60±0.13) and blank control group had no significant change(P＞0.05).Procollagen Ⅰ mRNA expression between different concentrations of Ghrelin and PDGF (3.11 ± 0.28,2.03 ±0.23,0.70 ± 0.06) was significantly reduced than PDGF group.The difference was statistically significant (P ＜0.05),and with the increase of the ghrelin concentration of the inhibition,the effect was more obvious in a concentration dependent manner.Western blot showed that α-SMA expression was lower in Ghrelin +PDGF group than PDGF group.AKT expression was higher in Ghrelin +PDGF group than PDGF group,indicating that PI3K-AKT may participate in the anti-fibrosis effect of ghrelin in LX2.After treatment of PI3K specific inhibitor,procollagen Ⅰ expression in LY294002+Ghrelin +PDGF group was significantly higher than Ghrelin +PDGF group((4.13±0.21) vs.(2.34±0.25),P＜0.05).Western blot also showed that α-SMA expression was higher in LY294002 + Ghrelin + PDGF group than Ghrelin + PDGF group.It was suggested that after inhibitation of PI3K,the anti-fibrosis effect of ghrelin in LX2 was attenuated.Conclusion After stimulated by PDGF in hepatic stellate cell,ghrelin can inhibit procollagen type Ⅰ and alpha-SMA synthesis in the process of hepatic fibrosis via PI3K-AKT pathway,thus,ghrelin may become one of the new ways of prevention and treatment of liver fibrosis.

Effect of rituximab on serum hepcidin, sCD40L, PDGF-BB and LDH levels in patients with malignant lymphadenoma / 中国生化药物杂志

Objective To explore the effect of rituximab on serum hepcidin (Hepc), soluble CD40 ligands (sCD40L), platelet derived growth factor-BB (PDGF-BB) and lactate dehydrogenase (LDH) in patients with malignant lymphadenoma.Methods 52 patients with malignant lymphoma from the hospital were randomly divided into control group and experimental group.The control group were treated by cyclophosphamide, pirarubicin, vincristine and prednisone and the experiment group were treated on the basis of control group with rituximab.The serum hepcidin, sCD40L, PDGF-BB and LDH levels were compared after treatment.Results Compared with pre-treatment, the serum hepcidin, PDGF-BB and LDH levels were lower( P<0.05), the sCD40L level was higher(P<0.05) post-treatment.The serum hepcidin, PDGF-BB and LDH levels in experimental group post-treatment were lower, sCD40L level was higher than those in control group(P<0.05).Conclusions The rituximab could reduce serum hepcidin, PDGF-BB and LDH levels and elevate serum sCD40L level in patients with malignant lymphadenoma.

Effect of hypoxia and PDGF-BB siRNA transfected on the production of PDGF-BB and proliferation in cultured human RPE cells / 中华实验眼科杂志

Background Proliferative vitreoretinopathy (PVR):no blood vessels,fibrous membrane cell proliferation occurs retinal surface.The development of specific mechanisms has not been completely clarified,but the retinal pigment epithelium(RPE) and platelet-derived growth factor(PDGF) for the past few years are the research hotspot.Objective To explore the effects of hypoxia on the production of PDGF-BB and proliferation in cultured human RPE (hRPE) cells.Methods In the experimental group,10,15,20,30,40 μmol/L CoCl2 were used to mimic hypoxia environment of hRPE cells,the control group did not add CoCl2 on cultured hRPE cells.By using reverse transcriptive PCR (RT-PCR) and enzyme linked immunosorbent assay(ELISA),the expression of PDGF-BB mRNA and protein was detected,and detected proliferation of hRPE cells by MTT.The cells were divided into 6 groups,different siRNAs were transfected into PDGF-BB siRNA1 group,PDGF-BB siRNA2 group,PDGF-BB siRNA3 group (because PDGF-BB has three different siRNAs,one of which is a valid siRNA),β-actin siRNA group,unrelated sequence siRNA group,only Lip2000 was plused in the blank control group.After transfection of 4-6 hours,except for blank control group,the other five groups were added to PDGF-BB mRNA and protein and its effect on the proliferation of hRPE cells obviously CoCl2 concentrations (15 μmol/L) simulated hypoxia environment of 24 hours,the detection of PDGF-BB mRNA and protein and hRPE cell proliferation rate.Results Neither PDGF-BB mRNA or protein was found in the blank control group.The production of PDGF-BB mRNA and protein in hRPE cells cultured by different concentrations were different,with significant differences among them (F=43.737,P＜0.01;F=54.612,P＜0.05),and the expressions of PDGF-BB mRNA and protein of 15 μmol/L CoCl2 group were more than other groups,with significant differences between the two groups (all at P＜0.05).MTT showed that PDGF-BB protein and hRPE cell proliferation rate in different concentions of CoCl2 groups were different,with significant differences among them(F=95.379,P＜0.01 ; F =63.375,P＜0.05),the expression of PDGF-BB protein was more and hRPE cell proliferation rate was higher in 15 μmol/L CoCl2group than other groups,with significant differences between the two groups (all at P＜0.05),postive correlation was found between PDGF-BB protein and hRPE cell proliferation rate (r =0.994,P＜0.05).The production of PDGF-BB mRNA and protein in the different siRNA transfected groups were different,with significant differences among them (F =156.330,125.650,both at P＜0.01),and the expressions of PDGF-BB mRNA and protein of PDGF-BB siRNA2 group were more than other groups,with significant differences between them(all at P＜0.05).MTT showed that PDGF-BB protein and hRPE cell proliferation rate in different siRNA transfectedgroups were different,with significant differences among them (F =73.131,98.564,both at P＜ 0.01).The expression of PDGF-BB protein was less and hRPE cell proliferation rate was lower in PDGF-BB siRNA2 group than other groups,with significant differences between them (all at P ＜ 0.05),postive correlation was found between PDGF-BB protein and hRPE cell proliferation rate (r =0.996,P＜0.05).Conclusions Proper hypoxia can induce the expression of PDGF-BB,PDGF-BB expression can significantly promote the proliferation of hRPE cells.In the transfection targeting PDGF-BB siRNA,PDGF-BB expression is suppressed,can effectively reduce the value of hRPE cell hyperplasia.

Artemisinin attenuates platelet-derived growth factor BB-induced migration of vascular smooth muscle cells

BACKGROUND/OBJECTIVES: Artemisinin (AT), an active compound in Arternisia annua, is well known as an anti-malaria drug. It is also known to have several effects including anti-oxidant, anti-inflammation, and anti-cancer activities. To date, the effect of AT on vascular disorders has not been studied. In this study, we investigated the effects of AT on the migration and proliferation of vascular smooth muscle cells (VSMC) stimulated by platelet-derived growth factor BB (PDGF-BB). MATERIALS/METHODS: Aortic smooth muscle cells were isolated from Sprague-Dawley rats. PDGF-BB stimulated VSMC migration was measured by the scratch wound healing assay and the Boyden chamber assay. Cell viability was determined by using an EZ-Cytox Cell Viability Assay Kit. The production of reactive oxygen species (ROS) in PDGF-BB stimulated VSMC was measured through H2DCF-DA staining. We also determined the expression levels of signal proteins relevant to ROS, including measures of extracellular signal-regulated kinase (ERK) 1/2 measured by western blot analysis and matrix metalloproteinase (MMP) 9 measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: AT (10 microM and 30 microM) significantly reduced the proliferation and migration of PDGF-BB stimulated VSMC in a dose-dependent manner. The production of ROS, normally induced by PDGF-BB, is reduced by treatment with AT at both concentrations. PDGF-BB stimulated VSMC treated with AT (10 microM and 30 microM) have reduced phosphorylation of ERK1/2 and inhibited MMP9 expression compared to untreated PDGF-BB stimulated VSMC. CONCLUSIONS: We suggest, based on these results, that AT may exert an anti-atherosclerotic effect on PDGF-BB stimulated VSMCs by inhibiting their proliferation and migration through down-regulation of ERK1/2 and MMP9 phosphorylation.