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Nonalcoholic fatty liver disease (NAFLD) has become one of the most common chronic liver diseases in the world, and it seriously harms human health. Recent studies have found that bone morphogenetic protein 4 (BMP4) might be associated with NAFLD. This article reviews the latest advances in the research on the association between BMP4 and NAFLD in China and globally and explores the potential mechanism of action of BMP4 on NAFLD, in order to provide new ideas for the prevention and treatment of NAFLD.
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Objective:To explore the effect of bone morphogenetic protein 4 (BMP4) on the glycolysis level of human retinal microvascular endothelial cells (hRMECs).Methods:A experimental study. hRMECs cultured in vitro were divided into normal group, 4-hydroxynonenal (HNE) group (4-HNE group) and 4-HNE+BMP4 treatment group (BMP4 group). 4-HNE group cell culture medium was added with 10 μmmol/L 4-HNE; BMP4 group cell culture medium was added with recombinant human BMP4 100 ng/ml after 6 h stimulation with 10 μmol/L 4-HNE. The levels of intracellular reactive oxygen species (ROS) were detected by flow cytometry. The effect of 4-HNE on the viability of cells was detected by thiazole blue colorimetry. Cell scratch test and Transwell cell method were used to determine the effect of 4-HNE on cell migration. The relative expression of BMP4 and SMAD9 mRNA and protein in normal group and 4-HNE group were detected by realtime quantitative polymerase chain reaction and Western blot. Seahorse XFe96 cell energy metabolism analyzer was used to determine the level of intracellular glycolysis metabolism in normal group, 4-HNE group and BMP4 group. One-way analysis of variance was used for comparison between groups.Results:The ROS levels in hRMECs of normal group, 4-HNE group and BMP4 group were 21±1, 815±5, 810±7, respectively. Compared with the normal group, the levels of ROS in the 4-HNE group and the BMP4 group were significantly increased, and the difference was statistically significant ( F=53.40, 50.30; P<0.001). The cell viability in the normal group and 4-HNE group was 1.05±0.05 and 1.28±0.05, respectively; the migration rates were (0.148±0.005)%, (0.376±0.015)%; the number of cells passing through the pores were 109.0±9.6, 318.0±6.4, respectively. Compared with the normal group, the 4-HNE group had significantly higher cell viability, cell migration rate, and the number of cells passing through the pores, and the differences were statistically significant ( F=54.35, 52.84, 84.35; P<0.05). The relative expression levels of BMP4 and SMAD9 mRNA in the cells of the 4-HEN group were 1.680±0.039 and 1.760±0.011, respectively; compared with the normal group, the difference was statistically significant ( F=53.66, 83.54; P<0.05). The relative expression levels of BMP4 and SMAD9 proteins in the cells of the normal group and 4-HEN group were 0.620±0.045, 0.860±0.190, 0.166±0.049, 0.309±0.038, respectively; compared with the normal group, the differences were statistically significant ( F=24.87, 53.84; P<0.05). The levels of intracellular glycolysis, glycolytic capacity and glycolytic reserve in normal group, 4-HNE group and BMP4 group were 1.21±0.12, 2.84±0.24, 1.78±0.36, 2.59±0.11, 5.34±0.32, 2.78±0.45 and 2.64±0.13, 5.20±0.28, 2.66±0.33. Compared with the normal group, the differences were statistically significant (4-HNE group: F=86.34, 69.75, 58.45; P<0.001; BMP4 group: F=56.87, 59.35, 58.35; P<0.05). There was no significant difference in intracellular glycolysis, glycolysis capacity and glycolysis reserve level between 4-HNE group and BMP4 group ( F=48.32, 56.33, 55.01; P>0.05). Conclusion:BMP4 induces the proliferation and migration of hRMECs through glycolysis.
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Abstract This study tested the hypothesis that head and neck radiotherapy (HNRT) impacts the immunoexpression of type I collagen, bone sialoprotein (BSP) and bone morphogenetic protein 4 (BMP4), thereby leading to micromorphological changes in the dentin-pulp complex (DPC), and promoting the onset and progression of radiation caries (RC). Twenty-two demineralized sections of carious teeth (a group of 11 irradiated teeth and a control group of 11 non-irradiated teeth) extracted from 19 head and neck cancer patients were analyzed by conventional optical microscopy and immunohistochemistry to investigate the micromorphology (cellular layer hierarchy, blood vessels, odontoblasts, fibroblasts, extracellular matrix, calcification, necrosis, reactionary dentin formation, and chronic inflammation), and the patterns of staining/immunolocalization of type I collagen, BSP and BMP4 in the dental pulp of irradiated and control samples. No significant differences attributable to the direct impact of radiotherapy were detected in DPC micromorphology between the groups. In addition, the patterns of immunohistochemical staining and immunolocalization of the proteins studied did not differ between the irradiated and the control samples for type I collagen, BSP or BMP4. This study rejected the hypothesis that HNRT directly damages dentition by changing the organic components and the microstructure of the DPC, ultimately leading to RC.
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Objective:To observe the effect of bone morphogenetic protein 4 (BMP4) on the proliferation and migration of human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:The hRMEC cultured in vitro were divided into control group, 4-hydroxynonenal (HNE) treatment group (4-HNE group), 4-HNE+BMP4 group (BMP4 group). Cell culture medium of 4-HNE treatment group was added with 10 μmmol/L 4-HNE; cell culture of BMP4 group was cultured with 10 μmmol/L 4-HNE, and after stimulation for 6 h, 100 ng/ml recombinant human BMP4 was added. The effects of 4-HNE and BMP4 on hRMEC viability was detected by thiazole blue colorimetric method. The effects of 4-HNE and BMP4 on cell migration was determined by cell scratch test. The relative expression of BMP4 mRNA in the cells of the control group and 4-HNE treatment group and the mRNA expression of the control group, the fibronectin (FN) of BMP4 group, laminin (Laminin), α-smooth muscle contractile protein (α-SMA), and collagen type Ⅰ (Collagen Ⅰ), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the relative expression of BMP4 protein in the control group and 4-HNE group. The control group and 4-HNE group were compared by t test. Results:Compared with the control group, cell viability ( t=12.73, 16.26, P=0.000 2, <0.000 1), cell migration rate ( t=28.17, 37.48, P<0.000 1, <0.000 1) in 4-HNE group and BMP4 group were significantly increased, and the difference was statistically significant; the relative expression of BMP4 mRNA and protein in the 4-HNE group was significantly increased, and the difference was statistically significant ( t=16.36, 69.35, P=0.000 1, <0.000 1). The qRT-PCR test results showed that compared with the control group, the relative expression of VEGF, FN, Laminin, α-SMA, Collagen Ⅰ, and CTGF mRNA in the cells of the BMP4 group was significantly increased, and the difference was statistically significant ( t=10.61, 17.00, 14.85, 7.78, 12.02, 10.61, P=0.0004, <0.000 1, 0.000 1, 0.001 5, 0.000 1, 0.000 4). Conclusion:BMP4 can induce the proliferation and migration of hRMEC; it can also regulate the expression of angiogenesis factors and fibrosis-related factors in hRMEC.
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Purpose To explore the expression of bone morphogenetic protein-4 (BMP-4) in prostate cancer tissues and to analyze the effects of its down-regulation on proliferation and invasion of prostate cancer cells. Methods The expression of BMP-4 protein in prostate tissues of 58 patients with prostate cancer and 40 patients with benign prostatic hyperplasia (BPH) was detected by immunohistochemical En Vision method. The BMP-4 protein expression of different prostate cancer cells (LNcap, PC-3, Du145) and BPH was analyzed by Western blot.BMP-4 siRNA and control siRNA were transfected into LNcap respectively and then divided into three groups: untreated group, control siRNA group and BMP-4 siRNA group. Western blot was used to detect the expression levels of BMP-4 protein, MMP-2and MMP-9 in LNcap cells among the three groups. The proliferation of three groups was detected by CCK 8. The invasion ability of three groups was detected by Transwell chamber. Results The positive proportion of BMP-4 protein in prostate cancer tissues was higher than that in benign prostatic hyperplasia (P < 0.05) , and the expression level of BMP-4 protein in three kinds of prostate cancer cells was higher than that in BPH cell. The expression of BMP-4 protein in BMP-4 siRNA group was lower than that in untreated group and control siRNA group.The cell proliferation ability and cell invasion ability in BMP-4siRNA group were lower than those in untreated group and control siRNA group. The expression levels of MMP-2 and MMP-9proteins in BMP-4 siRNA group were lower than those in untreated group and control siRNA group. Conclusion BMP-4 is highly expressed in prostate cancer tissues and is associated with proliferation and invasion of prostate cancer cells, and it may become a potential therapeutic target for prostate cancer.
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<p><b>OBJECTIVE</b>This study aimed to construct the expression of bone morphogenetic protein-4 (BMP4) lentiviral vector gene and explore its influence on the biological activity of mouse induced pluripotent stem (iPS) cells.</p><p><b>METHODS</b>iPS cell lines stably overexpressing BMP4 were constructed by lentivirus transfection (BMP4-overexpressing group). Cells without transfection served as the blank group, and cells with only vector transfection served as the empty-vector group. Cell proliferation was detected by CCK8, and the expression levels of ameloblastin (AMBN), cytokeratin (CK) 14, dentin sialophospho-protein (DSPP), bone sialoprotein (BSP), and Runx2 mRNA were detected by quantitative polymerase chain reaction. Alkaline phosphatase (ALP) activity was used to detect the degree of cell differentiation.</p><p><b>RESULTS</b>Compared with blank and empty-vector groups, proliferation activity and ALP activity of BMP4-overexpressing group obvious increased (P<0.05), BMP4, AMBN, CK14, DSPP, BSP, Runx2 mRNA expression also increased (P<0.05).</p><p><b>CONCLUSIONS</b>BMP4 can significantly promote the odontogenic differentiation of iPS.</p>
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Objective To observe the effect of bone morphogenetic protein 4(BMP4) on the infil-tration of macrophages and the expression of nuclear factor-κB in mice model of unilateral ureteral obstruc-tion(UUO).Methods C57BL/6 mice were randomly divided into four groups:injected intra-abdominally with saline-sham-operated group(saline-sham group,n =8),injection intra-abdominally with saline-UUO-operated group(saline-UUO group,n=8),injection intra-abdominally with anti-BMP4-sham-operated group (anti-BMP4-sham group,n=8),and injection intra-abdominally with anti-BMP4-UUO-operated group(anti-BMP4-UUO group,n=8).Either saline or anti-BMP4(group 200 μl/gram of body weight per day) were injected intra-abdominally for 7 days after surgery.Mice were sacrificed at 7th day to evaluate the expression of CD68 and p-P65 by immunohistochemical staining in each group.Besides,the p-P65 protein level was also analyzed by Western blotting in four groups.Results Immunohistochemical staining showed that the expres-sions of CD68 and p-P65 were significantly reduced in the kidney cortices in anti-BMP4-UUO group than in saline-UUO group(P<0.05,respectively).Similar to that,the p-P65 protein level was significantly reduced in the kidney cortices in anti-BMP4-UUO group than in saline-UUO group(P < 0.05,respectively). Conclusion BMP4 participates in the process of renal interstitial inflammation in obstructive nephropathy, and may play a role through the activation of nuclear factor-κB signaling pathway.
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ObjectiveTo explore regulation effect of miR-340-5p on regulating bone morphogenetic protein 4(BMP4) expression and the differentiation of rat's NSCs.MethodsNSCs of rats were divided randomly into three groups: normal group (Mock),group with nonsense oligonucleotide (Anti-Con) and group with antisense oligonucleotide of miR-340-5p (Anti-miR-340-5p).The qRT-PCR was used to detect the relative expression of miR-340-5p.The expression of NF200 and MAP-2 was detected by immunocytochemical staining and immunofluorescence,respectively.Western blot was used to detect the expression of BMP4 protein.ResultsThe relative levels of miR-340-5p expression were significantly decreased in Anti-miR-340-5p group (0.14±0.01) compared with that of Mock group(1.01±0.17) and Anti-Con group(1.07±0.13) (P<0.01).Immunocytochemical staining indicated that NF200 was positive in cells of Anti-miR-340-5p group.The proportion of MAP2 positive cells was increased in Anti-miR-340-5p group compared with other groups (P<0.05).Western blot showed the increased expression of BMP4 protein in Anti-miR-340-5p group (0.84±0.09) compared with Mock group(0.53±0.04) and Anti-Con group (0.63±0.09) (P<0.05).ConclusionThe miR-340-5p may exert a potential function in regulating differentiation of NSCs into neurons through a negative regulation of BMP4.
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Bone morphogenetic protein-4(BMP4) is one of BMPs member,it not only can play an important role in cell proliferation and invasion,but also play an important role in cell differentiation,apoptosis and angiogenesis.A study showed BMP4 in malignant tumor can inhibit the growth of tumor cells,and also can induce tumor cell invasion,metastasis and epithelial mesenchymal transformation,and SMAD is the classic signaling pathways.BMP4 is expected to become a potential therapeutic targets and in evaluating the prognosis,immune therapy in malignant tumor.In this paper,the structure and function of BMP4 and its mechanism in tumor were reviewed.
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Objective To investigate the mechanism of bone morphogenetic protein 4(BMP4) in the genesis of Barrett′s esophagus.Methods Human esophageal epithelial cell (HEEC) and MRC-5 were cultured.The effects of different concentration of BMP4 and different pH value of hydrochloric acid or glycocholic acid on the expression of caudal-related homeobox transcription factor 2(CDX2) in HEEC were detected by real time polymerase chain reaction.The effects of different pH value of hydrochloric acid or glycocholic acid on BMP4 expression in MRC-5 were also investigated.Independent sample t test was performed for statistical analysis.Results After HEEC stimulated by BMP4 at 0.1, 1.0, 10.0 and 100.0 ng/mL, the relative quantity expressions of CDX2 were 1.617±0.246, 2.489±0.455, 5.629±0.449 and 13.670±1.689, respectively, which were higher than those of control group (1.000±0.043, 1.029±0.094, 1.001±0.002 and 1.049±0.051, respectively), and the differences were statistically significant (t=2.47, 3.14, 10.31, 7.47;all P<0.05).After MRC-5 stimulated by acid at pH four or five, or glycocholic acid at pH four or five, the relative quantity expressions of BMP4 were 2.430±0.105, 2.394±0.145, 125.900±12.620 and 2.128±0.215, respectively, which were higher than those of control group(1.025±0.095, 0.999±0.007, 1.060±0.138 and 0.893±0.110,respectively), and the differences were statistically significant (t=9.94, 9.59, 9.89, 5.11;all P<0.01).Conclusion BMP4 can increase the expression of CDX2 in HEEC, which promote the genesis of Barrett′s esophagus.
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Objective To explore the reparation role of recombinant human bone morphogenetic protein-4 mature peptide (rhBMP-4m) on hematopoietic system injury induced by irradiation in mice.Methods Ninety BALB/c mice were randomly divided into normal control group,model group and rhBMP-4m treatment group,with 30 mice in each group.The mice in model group and rhBMP-4m treatment group were irradiated by 60Co γ-ray(7.5 Gy) for 200 s;while the mice in normal control group did not receive irradiation.After irradiation,the mice in model group were given physiological saline 1.0 mL by peritoneal injection per day for 6 days;the mice in rhBMP-4m treatment group were given rhBMP-4m 0.5 mg by peritoneal injection per day for 6 days.The peripheral white blood cell count and marrow mononuclear cells,the percent of CD34 + cells in marrow mononuclear cells of mice were detected on the first,third,fifth,seventh,ninth day after irradiation;the number of spleen nodus and the pleen weight/body weight ratio were detected at the ninth day after irradiation.Results There was no statistic difference in the number of peripheral white blood cells of mice among the three groups on the first day after irradiation(P ＞ 0.05).The peripheral white blood cell count of mice in model group was significantly lower than that in the normal control group on the third,fifth,seventh,ninth day after irradiation (P ＜ 0.01).There was no statistic difference in the number of peripheral white blood cells of mice between rhBMP-4m treatment group and model group on the third,fifth day after irradiation (P ＞ 0.05);the peripheral white blood cell count of mice in rhBMP-4m treatment group was significantly higher than that in model group on the seventh,ninth day after irradiation (P ＜ 0.05).The number of marrow mononuclear cells of mice in model group was significantly lower than that in the normal control group at each time piont after irradiation(P ＜ 0.05).There was no statistic difference in the number of marrow mononuclear cells of mice between the rhBMP-4m treatment group and model group on the first,third day after irradiation (P ＞ 0.05);the number of marrow mononuclear cells of mice in rhBMP-4m treatment group was significantly higher than that in the model group on the fifth,seventh,ninth day after irradiation(P ＜ 0.05).The proportion of CD34 + cells in mononuclear cells of mice in model group was significantly lower than that in the normal control group at each time piont after irradiation(P ＜0.01).There was no statistic difference in the proportion of CD34 + cells in mononuclear cells of mice between the rhBMP-4m treatment group and model group on the first,third day after irradiation (P ＞ 0.05);the proportion of CD34 + cells in mononuclear cells of mice in rhBMP-4m treatment group was significantly higher than that in the model group on the fifth,seventh,ninth day after irradiation(P ＜ 0.05).Compared with the normal control group,the number of spleen nodus was increased and the spleen weight/body weight ratio of mice was decreased in model group on the ninth day after irradiation(P ＜ 0.05);the number of spleen nodus and the spleen weight/body weight ratio of mice in rhBMP-4m treatment group were significantly higher than those in the model group on the ninth day after irradiation (P ＜ 0.01).Conclusion rhBMP-4m can accelerate the reconstruction of bone marrow hematopoietic system of mice with hematopoietic system injury induced by irradiation.
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@#Bone morphogenetic proteins(BMPs)belongs to the transforming growth factor beta(TGF-β)superfamily, which was isolated from the decalcified bone matrix by Urist in 1965.As one of the members of BMPs family, BMP4 not only promotes the differentiation and development of chondrocytes, but also plays an important role in the regulation of eye cells. BMP4 can mediate apoptosis of retinal vascular endothelial cells and inhibit angiogenesis, and is expressed in adult corneal epithelium, corneal cells and corneal endothelial cells. Also, participated in the layered corneal epithelium cells proliferation. In this paper, the recent advances in the study of BMP4 in ocular tissues are summarized, and the function of BMP4 in different developmental stages of eye is briefly reviewed.
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Objective Bone morphogenetic protein 4 ( BMP4) induces rat myocardial hypertrophy in H9c2 cells, but the spe-cific mechanism remains unclear .The study aimed to elucidate the role of autophagy in myocardial hypertrophy and its relationship with extracellular signal regulating kinase ( ERK1/2) signal transduction pathway . Methods H9c2 cardiomyocytes were randomly divided into 4 groups:control group, BMP4 group, BMP4+PD98059 (ERK1/2 signal pathway inhibitor) group and BMP4+3MA (autophagy inhibitor) group.With PD98059(50μmol/L) and 3MA(5mmol/L) blocking for 30min, BMP4 (50μg/L) were added.Expressions of tiny tube related proteins 1 light chain 3 (LC3) and p-ERK1/2 were detec-ted after culturing for 30min.48h later, measurements were made on cell surface area , average protein content and α-smooth muscle actin (α-SMA ) protein expression level .Cell morphology and size were measured respectively by inverted microscope and Image J software .Total protein content was detected by BCA , and western blot was used to measure the expressions of LC3, ERK1/2, p-ERK1/2 and α-SMA. Resul ts 30min later, the expressions of LC3 and p-ERK1/2 were significantly higher in BMP4 group than in control group [(1.54 ±0.05) vs (1.95 ±0.11),(0.94 ±0.04) vs (1.33 ± 0.06),P0.05). Conclusion Autophagy may participate in BMP4-induced rat myo-cardial hypertrophy in H9c2 cells through ERK1/2 signal transduction pathway .The clinical treatment of myocardial hypertrophy may benefit from the blocking of autophagy or ERK 1/2 signal transduction pathway .
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Objective:To explore the role of bone morphogenetic protein 4 (BMP-4) in hepatic progenitor cells (HPCs).Methods: The effect of BMP-4 on rat hepatic oval cells was examined by using the WB-F344 rat hepatocytic epithelial stem-cell-like cell line. This hepatocytic cell line could exert various hepatocyte functions including the secretion of albumin and urea. Immunohistochemistry was used to examine the effects of BMP-4 and its antagonist, Noggin, on the proliferation and differentiation of these cells, cellular uptake and excretion of indocyanine green, the periodic acid-schiff (PAS) assay for glycogen storage and the expression of hepatic markers.Results:Our results showed for the first time that BMP-4 may acted as a potential inducer of hepatic differentiation in rat hepatic oval cells.Conclusions:This cell source offers a much-needed attractive and expandable source for future investigations of drug screening, stem cell technologies and cellular transplantation, in a society with increasing levels of liver disease and damage.
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OBJECTIVE@#To explore the role of bone morphogenetic protein 4 (BMP-4) in hepatic progenitor cells (HPCs).@*METHODS@#The effect of BMP-4 on rat hepatic oval cells was examined by using the WB-F344 rat hepatocytic epithelial stem-cell-like cell line. This hepatocytic cell line could exert various hepatocyte functions including the secretion of albumin and urea. Immunohistochemistry was used to examine the effects of BMP-4 and its antagonist, Noggin, on the proliferation and differentiation of these cells, cellular uptake and excretion of indocyanine green, the periodic acid-schiff (PAS) assay for glycogen storage and the expression of hepatic markers.@*RESULTS@#Our results showed for the first time that BMP-4 may acted as a potential inducer of hepatic differentiation in rat hepatic oval cells.@*CONCLUSIONS@#This cell source offers a much-needed attractive and expandable source for future investigations of drug screening, stem cell technologies and cellular transplantation, in a society with increasing levels of liver disease and damage.
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Objective: To explore the role of bone morphogenetic protein 4 (BMP-4) in hepatic progenitor cells (HPCs). Methods: The effect of BMP-4 on rat hepatic oval cells was examined by using the WB-F344 rat hepatocytic epithelial stem-cell-like cell line. This hepatocytic cell line could exert various hepatocyte functions including the secretion of albumin and urea. Immunohistochemistry was used to examine the effects of BMP-4 and its antagonist, Noggin, on the proliferation and differentiation of these cells, cellular uptake and excretion of indocyanine green, the periodic acid-schiff (PAS) assay for glycogen storage and the expression of hepatic markers. Results: Our results showed for the first time that BMP-4 may acted as a potential inducer of hepatic differentiation in rat hepatic oval cells. Conclusions: This cell source offers a much-needed attractive and expandable source for future investigations of drug screening, stem cell technologies and cellular transplantation, in a society with increasing levels of liver disease and damage.
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Objective To investigate the effect of umbilical cord blood monocytes(UCBMC) transplantation in neonatal rats with hypoxic-ischemic brain damage (HIBD) on rat astrocyte proliferation and its correlation with bone morphogenetic protein(BMP) 4.Methods Forty 7-day-old SD rats with the random number table,were divided into normal control(CON) group,hypoxic-ischemic(HI) group,normal(N) + UCBMC group,HI + UCBMC group,10 rats in each group.HIBD model was prepared according to the Rice method.Twenty-four hours after hypoxia,the UCBMC group and HI + UCBMC group were injected with 3 × 106 UCBMC via the lateral ventricle.Seven days after transplantation,changes in the number of neurons were observed by Nissl staining;the expression of glial fibrillary acidic protein (GFAP) was observed by Western blot;the astrocyte proliferation was observed by proliferating cell nuclear antigen (PCNA)/GFAP,BMP4/GFAP immunofluorescence double staining,and their correlation was analyzed.Results Nissl staining showed that the neurons at cerebral cortex and hippocamp were irregularly arranged and decreased in the HI group,and the number of Nissl-stained cells were significantly less than that in the CON group(tcortex =26.54,thippocamp =32.26,all P ＜0.05) ;but the Nissl-stained cells were well arranged in the HI + UCBMC group and more Nissl-stained cells were observed as compared with the HI group (tcortex =10.18,thippocamp =12.56,all P ＜ 0.05) ; Western blot showed that the expression of GFAP in the HI group was significantly higher than that in CON group(t =5.50,P ＜ 0.05) ;but the expression of GFAP in the HI + UCBMC group was significantly lower than that in the HI group (t =3.04,P ＜ 0.05) ; immunofluorescence double staining showed that there were more PCNA + GFAP + cells in the HI group compared with the CON group(t =10.39,P ＜ 0.05),but fewer PCNA + GFAP + cells were observed in the HI + UCBMC group than those in the HI group(t =3.72,P ＜ 0.05).And there were more BMP4 + cells in the HI group compared with the CON group (t =5.52,P ＜ 0.05),but fewer BMP4 + cells were observed in the HI + UCBMC group than those in the HI group(t =2.33,P ＜0.05).The fluorescence intensity of GFAP were correlated with that of BMP4 in HI + UCBMC group (r =0.84,P ＜ 0.05).Conclusions UCBMC transplantation can decrease the proliferation of the astrocytes,thus promote brain damage repair and its mechanism might be collected with the decreased expression of BMP4.
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Objective To investigate the changes of plasma bone morphogenetic protein-4 (BMP-4) levels in patients with coronary heart disease (CHD) and rosuvastatin intervention effect on BMP-4 level.Methods Fifty-two patients with CHD and 35 health people were enrolled in this study as CHD group and control group.ELISA method was used to detect the concentration of plasma BMP-4.Analyzed the relationship between plasma BMP-4 and blood lipids,flow-mediated dilation (FMD),nitric oxide (NO),cyclooxygenase-2 (COX-2),malondialdehyde (MDA) and superoxide dismutase (SOD).And observed the changing of plasma BMP-4 before and after rosuvastatin intervention.Results Plasma BMP-4 level in CHD patients was (7.53 ± 1.20) μg/L,higher than that of control group ((3.81 ± 0.79) μ g/L,t =3.541,P =0.006).After rosuvastatin treatment,plasma BMP-4 level in CHD patient was decreased from (7.53 ± 1.20) μg/L to (5.40± 0.98) μg/L (t =1.436,P =0.001).Plasma BMP-4 level was positively correlated with COX-2,MDA,low-density lipoprotein cholesterol,total cholesterol (r =0.395,0.350,0.274,0.288 respectively,P ＜ 0.01 or P ＜0.05).But,it was negatively correlated with NO,high-density lipoprotein cholesterol,SOD,FMD (r =-0.291,-0.253,-0.476,-0.320 respectively,P ＜0.01 or P ＜0.05).COX-2,SOD and FMD were independent risk factors of plasma BMP-4 in patients with CHD.Conclusion Oxidative stress and endothelial dysfunction are in patients with CHD.Rosuvastatin treatment can remarkably reduce plasma BMP-4 level,alleviate vascular endothelium injury induced by oxidative stress and improve endothelial function in patients with CHD.
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Background & objectives: BMP (bone morphogenetic protein)-4/7 and bFGF (basic fibroblast growth factor) significantly promote the osteogenic activity and the proliferation of rabbit BMSCs (bone marrow stromal cells), respectively. However, their synergistic effects on the proliferation and the differentiation of BMSCs remain unclear. In the present study, the effects of bFGF and BMP-4/7 were investigated on the proliferation and the differentiation of rat BMSCs in vitro. Methods: BMSCs were isolated from New Zealand white rabbits and cultured to the third passage. The samples were divided into five groups according to the material implanted: (A) 80 ng/ml BMP-4/7; (B) 80 ng/ml bFGF; (C) 30 ng/ml BMP-4/7 and 30 ng/ml bFGF; (D) 50 ng/ml BMP-4/7 and 50 ng/ml bFGF; and (E) 80 ng/ml BMP-4/7 and 80 ng/ml bFGF. Cell proliferation was analyzed using methyl thiazolyl tetrazolium (MTT) assay. Alkaline phosphatase activity and osteocalcin (OC) dynamics were also measured. Results: BMP-4/7 alone significantly (P<0.05) promoted the proliferation of BMSCs. At the same time, it also promoted or inhibited the osteogenic differentiation of BMSCs. The synergistic effects of BMP-4/7 and bFGF significantly promoted both the proliferation and the osteogenic differentiation of BMSCs. The treatment of the synergistic effects was dose and time dependent. Interpretation & conclusions: A rational combination of BMP-4/7 and bFGF can promote the proliferation and the osteogenic differentiation of BMSCs. In addition, the synergistic functions are effective.
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Bone morphogenetic protein type 2 (BMP-2) is a potent local factor, which promotes bone formation and has been used as an osteogenic supplement for mesenchymal stem cells. OBJECTIVES: This study evaluated the effect of a recombinant BMP-2 as well as the endogenous BMP-4 and BMP-7 in the osteogenic differentiation of adipose-derived stem cells (ASCs) in medium supplemented with ascorbate and β-glycerophosphate. MATERIAL AND METHODS: Human ASCs were treated with osteogenic medium in the presence (ASCs+OM+BMP-2) or absence (ASCs+OM) of BMP-2. The alkaline phosphatase (ALP) activity was determined and the extracellular matrix mineralization was evaluated by Von Kossa staining and calcium quantification. The expressions of BMP-4, BMP-7, Smad1, Smad4, and phosphorylated Smad1/5/8 were analyzed by western blotting. Relative mRNA expressions of Smad1, BMP receptor type II (BMPR-II), osteonectin, and osteocalcin were evaluated by qPCR. Results: ASCs+OM demonstrated the highest expression of BMP-4 and BMP-7 at days 21 and 7, respectively, the highest levels of BMPR-II mRNA expression at day 28, and the highest levels of Smad1 mRNA at days 14 and 28. ASCs+OM+BMP-2 demonstrated the highest levels of Smad1 mRNA expression at days 1, 7, and 21, the highest expression of Smad1 at day 7, the highest expression of Smad4 at day 14, the highest ALP activity at days 14 and 21, and expression of phosphorylated Smad1/5/8 at day 7. ASCs+OM and ASCs+OM+BMP2 showed similar ALP activity at days 7 and 28, similar osteonectin and osteocalcin mRNA expression at all time periods, and similar calcium depositions at all time periods. CONCLUSIONS: We concluded that human ASCs expressed endogenous BMP-4 and BMP-7. Moreover, the supplementation of ASCs with BMP-2 did not increase the level of osteogenic markers in the initial (ALP activity), intermediate (osteonectin and osteocalcin), or final (calcium deposition) phases, suggesting that the exogenous addition of BMP-2 did not improve the in vitro osteogenesis process of human ASCs.