肝细胞生长因子激活胶原降解途径逆转心肌纤维化

Objective To study the effects of hepatocyte growth factor (HGF) on angiotensin Ⅱ(Ang Ⅱ )-induced synthesis and degradation of cardiac fibroblast collagen in neonatal mice.Methods Cardiac fibroblasts (CFs) of neonatal Sprague-Dawley (SD) rats were isolated by differential adhesion, and were identified by immunocytochemistry.The cells were assigned to normal control group (group C) , 10-6 mol/L Ang Ⅱ group (group A), 10μg/L HGF + 10-6 mol/L Ang Ⅱ group (group H1) and 100μg/L HGF +10-6 mol/L Ang Ⅱ group (group H2).All the groups were treated for 48 h.Hydroxyproline assay was used to determine the collagen protein level.RT-PCR was used to detect the expression of ColⅠ, ColⅢ, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-l) mRNA, and Western blotting was employed to measure Col I protein.In addition, MMP-1 activity was evaluated by collagenase Ⅰ activity assay kit.Results After intervention for 48 h, the collagen protein levels in groups A and H1 were significantly higher compared with group C [ (39.08±2.71) mg/L vs (37.45±4.22) mg/L vs (23.73 ±±1.62) mg/L, all P＜0.05] and the collagen protein level in group H2 [(26.03±3.04) mg/L] was lower than those of groups A and Hl(P＜0.05), but was not statistically different from group C.The expression levels of Col Ⅰ ,Col M and TIMP-1 showed an order of group A＞group HI＞group H2> group C (all P＜0.05), and for MMP-1 mRNA, group A ＜ group H1 ＜ group H2 ＜ group C (all P＜0.05).The levels of Col I protein expression showed an order of group A＞group HI＞group H2＞group C (89.90±4.29 vs 68.21?1.43 vs 36.08?.8 vs 30.14?.36, all P＜0.05).The expression and activity of MMP-1 mRNA in group A and H1 were obviously lower than those in group C (all P＜0.05), while group H2 were comparable to group C in these data.Conclusion HGF re-regulates MMP-1-TIMP-1 balance and ameliorates myocardiac fibrosis mainly by activating collagen degradation.     

EGFL7基因沉默对裸鼠乳腺癌血管生成的影响

Objective To investigate the effect of short hairpin RNA (shRNA) targeting at epidermal growth factor-like domain 7 (EGFL7) gene on angiogenesis of breast carcinoma and its mechanism in nude mice. Methods shRNA targeting at EGFL7 gene was constructed and transfected into SGC-7901 cells (pshEGFL7 group) , meanwhile , the cells transfected with vector plasmids were as a control group. Positive clones were selected and the transplanted tumor animal models constructed in nude mice , and the growth and volume of tumors were observed. After 8 weeks , EGFL7 mRNA and protein in transplanted tumor tissues were detected,and graded. Moreover, Anti-CD34, VEGF and TSP1 were stained by the immuno- chemistry method,and MMP-2 and TIMP2 mRNA were detected by RT-PCR. Results EGFL7 mRNA was down regulated significantly in the pshEGFL7 group. In the psh EGFL7 group, the tumor volume was ( 1.86 ± 0. 65) cm3, MVD was 20. 84 ± 6.38; while in the control group , tumor volume was (4.86 ± 1.15) cm3, MVD was 39.48 ± 9.01, In the EGFL7 group ,TSP1 protein presented positive , and VEGF protein presented weakly positive or negative.The expression of MMP-2 mRNA decreased ,TIMP2 mRNA increased in the pshEGFL7 group,and there were significant differences compared with the control group , P ＜ 0.01. Conclusion RNA interference targeting at EGFL7 gene can balance TSP1/VEGF, through regulating the expression of MMP-2/TIMP2 to impair angiogenesis of breast cancer in nude mice.     

Impact of CCAAT enhancer binding protein α on the differentiation and apoptosis of acute myeloid leukemia HL60 cells

Objective To investigate the effect of CCAAT enhancer binding protein α(C/EBPα) on differentiation and apoptosis in the acute myeloid leukemia HL60 cells in vitro and in vivo and its possible mechanism. Methods The C/EBPα expression plasmid pEGFP-C/EBPα and empty control plasmid were respectively transfected into HL60 cells with cationic liposome as transfected group and empty plasmid transfected group, and untreated HL60 cells served as control group. The cells stably expressing the C/EBPα gene were obtained by G418 selection. The morphological changes were observed under light microscope following WrightGiemsa staining. MTT assay was employed to evaluate cell proliferation, and flow cytometry(FCM) was performed to analyze cell apoptosis. Meanwhile, the expression of c-myc was respectively detected by RT-PCR and Western blot both at the mRNA and protein level. Twenty BALB/c nude mice were divided into 3 groups in a completely randomized design: 7 mice in transfected group, 7 mice in empty plasmid transfected group and 6 in control group. Three kinds of cells including pEGFP-C/EBPα-HL60 cells, pEGFP -HL60 cells and the control HL60 cells were injected into mice separately through the subcutaneous. The mice were sacrificed at 20 d after injection. The mass and size of subcutaneous xenograft tumors were measured and the cell apoptosis of subcutaneous tumor were detected by TUNEL. Results The pEGFP-C/EBPα-HL60 cell line stably expressing the C/EBPα gene was screened out. Compared to either empty plasmid transfected group or control group, the expression of C/EBPα could promote cellular differentiation of HL60. FCM showed higher apoptotic rate in transfected group[ (21.9±4.5)%,P＜0.05 ] ,while (5.4±1.4)% in control group and (5.0±1.3)% in empty plasmid transfected group. c-myc expression was significantly down-regulated by C/EBPα both at the mRNA and protein level. The mass and size of tumors in transfected group were smaller than those in empty plasmid transfected group and control group [ (5.35±1.12)g and(25±4)mm in control group, (5.12±1.31)g and ( 18±3)mm in empty plasmid transfected group ,while (3.26±0.72)g and ( 11±2)mm in transfected group, all P＜0.05]. More apoptosis cells were found in subcutaneous tumor of transfected group(both P＜0.05). Conclusion C/EBPα can not only inhibit the proliferation, but also induce massive apoptosis of HL60 cells, meanwhile C/EBPα is a tumor suppressor of acute myeloid leukemia.     

姜黄素抑制大鼠创伤性骨关节炎模型中软骨细胞核因子κB P65核转位的实验研究

BACKGROUND: Mechanical, inflammatory, and biochemical factors, particularly matrix metalloproteinases and reactive oxygen lead to chondrocyte degeneration in osteoarthritis. Curcumin has been shown to be a potent antioxidant; however, its protective effects against chondrocyte degeneration in osteoarthritis remain unclear. OBJECTIVE: To investigate the potential molecular mechanisms underlying the protective effects of curcumin on articular cartilage of osteoarthritis in rats. METHODS: A total of 30 Sprague-Dawley rats were used and randomly divided into model group (positive control, n=15) and normal group (negative control, n=15). Rat models of traumatic osteoarthritis were established, and then cartilage cells were isolated from articular cartilage and cultured in vitro. Chondrocytes were treated with curcumin (curcumin group) or PDTC (an inhibitor of nuclear factor-kappa B) for 24 hours. The expression level of nuclear factor-kappa B P65 in nucleus and cytoplasm in chondrocytes were determined by western blot assay and immunofluorescence. Moreover, mRNA expressions of type II collagen, matrix metalloproteinase-1 and -13 were analyzed using RT-qPCR. RESULTS AND CONCLUSION: Nuclear factor-kappa B P65 protein was mainly expressed in nucleus, but few in cytoplasm in positive control group; the reversed results were found in the curcumin group. Nuclear translocation of nuclear factor-kappa B P65 was observed mainly in nucleus in the positive control group; however, that was observed mainly in cytoplasm in the negative control, curcumin, and PDTC groups. Matrix metalloproteinase-1 and -13 mRNA expressions were significantly decreased, while type II collagen mRNA expression was significantly increased in the curcumin group compared with the positive control group. These findings indicated that curcumin protect chondrocytes against degeneration through inhibiting the activation of nuclear factor-kappa B signaling pathway, suppressing nuclear translocation of nuclear factor-kappa B P65 and inhibiting the expressions of matrix metalloproteinase-1 and -13, which are responsible for upregulation of type II collagen expression. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Effects of Tensile Stimulation and Electrical Stimulation on the Connexion-43 m RNA Levels of Cardiomyocytes in vitro

The heart is regulated by excitation-contraction mechanism and electrical stimulation, so both cardiac tissues and myocytes in vivo are affected by the mechanical and electrical factors at the same time. The device that can supply both tensile stimulation and electrical stimulation was produced. Cardiomyocytes were isolated by trypsin and collagenase Ⅱ combined digestion. After 72 h's culture, different electrical stimulations(0.5 V/cm, 1 V/cm, 2 V/cm; 1 Hz; 2 ms; 30 min) and mechanical stretching force(tensile 10%, 6 h) were used to stimulate cultured cardiomyocytes respectively. Then the combinative stimulation of mechanical and electrical stimulation with 0.5 V/cm was used to apply to cardiomyocytes simultaneously. After stimulation,Connexin-43(Cx-43) mRNA level was detected by RT-PCR. The stimulation by electrical pulse of 0.5 V/cm resulted in obvious enhancement of Cx-43 m RNA level compared to the control group and the Cx-43 mRNA levels at 0.5 V/cm were higher significantly than those under both 1.0 V/cm and 2.0 V/cm. After 10% of tensile stimulation for 6 h, expressions of Cx-43 mRNA were also significantly promoted. The combinative effect of mechanical and electrical stimulation on the expression of Cx-43 mRNA is higher than that under unique tensile stimulation or electrical stimulation,which is more beneficial to the development and connection of cardiomyocytes.     

红细胞生成素对大鼠阿霉素性心肌病的预防

Objective To investigate the preventive effect and potential mechanisms of erythropoietin (EPO) against doxorubicin (DOX)-induced cardiomyopathy. Methods Thirty-one Wistar rats were randomly divided into DOX group (n=12) , DOX + EPO group (n=11) and control group (n=8). Dilated cardiomyopathy was induced by intraperitoneal injection of DOX for both DOX group and DOX+EPO group, and normal saline or EPO was administered before DOX injection for preventive purpose. Left ventricular systolic function was evaluated by invasive haemodynamic measurements among three groups. Rats were then sacrificed for Masson-stained histopathology observation and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) analysis of apoptosis, with immunological detection for Bax and Bcl-2 by Western blotting. Results There was significant difference in left ventricular systolic function between DOX group and DOX+EPO group. The area ratio of myocardial fibrosis was significantly decreased from (12.14±1.07)% in the DOX group to (7.49±1.11)% in the DOX+EPO group (P<0.05). The apoptotic index was (0.93±0.08)% and (0.16±0.04)% in the DOX group and DOX + EPO group (P<0.05) , respectively. Western blotting showed that Bcl-2 protein expression was decreased in DOX group compared to control group, but significantly increased in the DOX + EPO group compared to DOX group (P<0.05). Conclusions EPO can protect against DOX-induced cardiomyopathy via anti-apoptotic pathways. The up-regulation of Bcl-2 protein expression may contribute to the protection.     

Regional myocardial heat-shock protein (HSP70) concentrations under different blood flow conditions

 The concentration of heat-shock proteins of 70 kD (HSP70) in heart tissue has been shown to increase during transient myocardial ischaemia and to persist during several hours of reperfusion. In this study the relationship between the local myocardial HSP70 concentration and blood flow was addressed for control physiological conditions and acute myocardial ischaemia. A specific aim of this study was to address the question of whether low flow areas under control physiological conditions have undergone a transient ischaemia during the preceding hours and thus may be in a state of hibernation or stunning. In 12 anaesthetized, open-chest beagle dogs (6 control and 6 with 60-min coronary artery stenosis) heart rate, mean aortic pressure, mean arterial partial pressure of O₂ and partial pressure of CO₂ averaged 85±16 beats/min, 94±14 mmHg, 102±17 mmHg and 39±6 mmHg, respectively. Regional HSP70 and myocardial blood flow (RMBF) were measured using an HSP70-enzyme-linked immunosorbent assay and the tracer microsphere technique, respectively, in samples of 250 mg wet mass. In the control group the mean RMBF was 1.06±0.59 ml·min^–1·g^–1 and the local HSP70 concentration was 7.08±1.03 μg/mg cytosolic protein. Myocardial HSP70 showed a blood flow-independent regional biological heterogeneity, equivalent to a coefficient of variation of 0.31. Local HSP70 concentrations did not differ (P>0.05) between control low and high flow samples, 6.16±1.0 vs 6.08±0.75 μg/mg cytosolic protein, respectively. However, after 60 min of coronary artery occlusion the local HSP70 concentration increased from 7.08 ±1.03 to 13.43±3.19 μg/mg cytosolic protein (P<0.001). There was a significant inverse relationship between the percent reduction of local blood flow and HSP70 (r=–0.56, P<0.001). From these results it is concluded that: (1) low flow samples under control physiological conditions are unlikely to be in a state of hibernation or stunning since their HSP70 concentration is normal and (2) the increase in the local HSP70 concentration during myocardial ischaemia reflects the degree of impairment of O₂ delivery. Received: 29 May 1998 / Received after revision: 14 August 1998 / Accepted: 25 August 1998     

hIL-24基因通过调控转化生长因子β/Smad通路影响瘢痕疙瘩的生物学特性

BACKGROUND:hIL-24, a tumor suppressor gene, can stimulate immune responses, inhibit the growth of tumor cells, and the formation of tumor vessels, and induce cell apoptosis. OBJECTIVE: To explore the effects of hIL-24 gene on the proliferation and apoptosis of fibroblasts in the keloid and the underlying mechanisms. METHODS: All the keloid specimens collected from 13 patients were used for fibroblast culture and indentification. Fibroblast of the keloid was transfected with or without hlL-24 lentivirus. Subsequently, mRNA expressions of transforming growth factor-β, Smad3, proliferating cell nuclear antigen, matrix metalloproteinase-2, -9, and metallopeptidase inhibitor 1 were determined. RESULTS AND CONCLUSION:Immunofluorescent staining and flow cytometry showed that vimentin antibody was expressed positively in cytoplasma of fibroblast cultures, and the purity was more than 97.8%. Western blot assay showed that hIL-24 expression was significantly increased in the transfected fibroblasts. Quantitative PCR showed that the overexpression rate of hIL-24 in fibroblasts was 81.7% and mRNA expressions of transforming growth factor-β, Smad3, proliferating cell nuclear antigen, matrix metalloproteinase-2, and -9 were significantly decreased, while metallopeptidase inhibitor 1 mRNA expression was significantly increased in hIL-24 transfection group compared with control group (P < 0.05). These findings suggest that hIL-24 gene inhibits the expressions of proliferating cell nuclear antigen, matrix metalloproteinase-2, and -9 in fibroblasts, and the underlying mechanism may involves TGF-β/Smad3 pathway. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

低强度脉冲式超声对关节软骨的修复

BACKGROUND: Articular cartilage injuries can result from a variety of causes. Conventional therapy cannot obtain the optimal clinical results. Low-intensity pulsed ultrasound has been shown to promote the repair of injured articular cartilage. OBJECTIVE: To investigate the effects of low-intensity pulsed ultrasound on the repair of injured articular cartilage. METHODS: Twenty New Zealand white rabbits were used to establish knee arthritis models and equally randomized into study and control groups, respectively. Rabbits in the study group received low-intensity pulsed ultrasound treatment, and sham low-intensity pulsed ultrasound treatment was given in the control group. At 8 weeks after treatment, pathological change and histological scores in articular cartilage tissue collected from both groups were determined. Moreover, the ultrastructure and type II collagen expression of chondrocytes were determined. Matrix metalloproteinase-13 mRNA expression was detected by quantitative real-time PCR. RESULTS AND CONCLUSION: At 8 weeks after treatment, toluidine blue staining showed a disordered arrangement of cells, decreased number of cartilage cells in each layer and cluster in the control group. Light disordered arrangement of cells, decreased appearance of the superficial layer cells and the cluster phenomenon were observed in the study group. Articular cartilage tissue scores were significantly decreased in the study group compared with the control group (P < 0.05). The chondrocytes were small, enlarged intracellular mitochondria and rough endoplasmic reticulum, cytoplasmic swelling, collagen fibrils coarse, well developed Golgi apparatus, and nuclear fragmentation were observed in the control group. In addition, the normal structure of organelles disappeared and cell degeneration was observed in the control group. In the study group, the size of chondrocytes and the Golgi complex and other organelles were normal, and the protein polysaccharide granules were observed in the cytoplasm and membrane. The mRNA expression of matrix metalloproteinase-13 in the study group was significantly lower than that in the control group (P < 0.05). Type II collagen immunoreactivity in the study group was stronger than that in the control group. No incision infection, suppuration, red swelling appeared in all rabbits. Our results suggest that low-intensity pulsed ultrasound can be used for the treatment of articular cartilage injury by alleviating the degradation of collagen type II and inhibiting the expression of matrix metalloproteinase-13. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

多聚左旋赖氨酸协同超顺磁性氧化铁标记大鼠C6脑胶质瘤细胞

Objective To investigate the labeling rate with poly-L-lysine (PLL) and super paramagnetic iron oxide particles (SPIO) for C6 rat glioma cells and the impact of PLL and SPIO on labeled cell bioactivity. Methods C6 rat glioma cells were incubated without SPIO or PLL (control group), with 25 mg/L SPIO (group A), 25 mg/L SPIO+0.75 mg/L PLL (group B), and 50 mg/L SPIO+1.5 mg/L PLL (group C) , respectively. The labeled cell bioactivity was analyzed by MTS assay (mono-nuclear cell direct cvtotoxicity assay) at different incubation time (6, 24 and 48 h) after treatment, and the labeling rate was detected by Prussian Blue staining. The labeled cells mass were imaged in vitro using a 3.0 T MRI scanner with GRE/30° T2*WI sequence. The R2* values and signal intensity were compared among these groups. Results Up to 48 h post-labeling, there was no significant decrease of cell bioactivity in all labeled cell groups (all P>0.05). Prussian Blue staining showed that the labeling rate was >98% in group B and C compared with about 70% in group A. The staining appeared increasingly darker in SPIO/PLL mixed labeled cells along with higher concentration of SPIO. 3.0 T MRI was signal-sensitive for the labeled cells. The R2* values were 11.76±5.74, 12.13±4.39, 61.22±27.85 and 90.07±35.59 for control group and group A, B and C, respectively. Difference of R2* value was not found between control group and group A (P>0.05), but was found between group B and C, meanwhile R* values of group B and C were significantly different with control group and group A (all P<0.01). Conclusions PLL may enhance the efficiency of SPIO labeling for C6 glioma cells and has no significant impact on the cell bioactivity. 3.0 T MRI with GRE/30° T2*WI sequence is signal-sensitive for labeled cells in vitro. The R2* value may increase along with intracellular SPIO level. 25 mg/L SPIO+0.75 mg/L PLL may yield satisfactory labeling for rat C6 glioma cells in vitro.     

多孔复合材料PHBV-SGBG的体外细胞相容性和体内成骨性

Objective To study the cytocompatibility of poly (3-hydroxybutyrate-co-3 -hydroxyvalerate)/sol-gel bioactive glass (PHBV/SGBG)composite with bone marrow stromal cells in vitro, and the in vivo osteogenesis efficiency of PHBV/SGBG composite under periosteal flap in bone defect animal models. Methods The cytocompatibility of PHBV/SGBG composite was tested by MTT assay and direct contact assay. Of established tibial defect canine models, PHBV/SGBG composite was embedded into the defects of the experimental group, while no treatment was given to the control group. Samples were harvested at weeks 2,4, 8, and 12 post-surgery. The effect of osteogenesis was inspected by scanning electron microscope observation and energy-dispersive X-ray analysis. Results The cytotoxicity score of PHBV/SGBG composite in the serial dilution extract and different culture time ranked from grades 0 to 1. Marrow stem cells (MSCs) attached to and then adhered firmly on the surface of PHBV/SGBG composite, and proliferated rapidly, with obvious cell prominences stretching into the micro-pores structure. In the experimental group, formation of new bone islands was observed at week 2 post-surgery, and mature bone regeneration at week 12. Scanning electron microscopy and energy-dispersive X-ray analysis showed increasing calcium peak in the experiment group. At week 4 Ca/P ratio was 1.086±0.034 in experiment group and 0.793±0.053 in control group. At week 12 Ca/P ratio was 1.603±0.067 in experiment group and 1.456±0.036 in control group (P<0.05). Conclusions PHBV/SGBG shows no cytotoxicity to MSCs, and possesses good osteo-conductibility and regeneration capability. It may be a promising composite for bone tissue engineering.     

雌激素受体β基因沉默对人成骨细胞转化生长因子β1和骨形态发生蛋白2表达的影响

BACKGROUND: There are few studies concerning estrogen receptor β gene, and its mechanism of regulating the bone metabolism is still unclear now.  OBJECTIVE: To analyze the effect of estrogen receptor β (ER β) silencing on the expressions of transforming growth factor β1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) in human osteoblasts METHODS: There were three groups: blank control group (hFOB 1.19 uninfected with any retrovirus); negative control group (containing invalid interference fragment ER β-shRNA-nc); optimal RNAi group (ER β-shRNA-3). ER β-shRNA retroviral vectors in the optimal RNAi group were used to transfect human osteoblasts followed by resistance screening and cell expansion. MTT assay was used to detect the proliferative activity of ER β-silenced osteoblasts. Then under estrogen intervention, the stable inhibition rate of ER β was determined using western blot assay, and the expressions of TGF-β1 and BMP-2 in human osteoblasts after ER β silencing were detected by RT-PCR technology and western blot assay. RESULTS AND CONCLUSION: Human osteoblasts that were stably transfected by ER β-shRNA-3 retroviral vector was selected successfully, and ER β silencing had no significant influence on the cell proliferation (P > 0.05). Under the interference of estrogen, the silencing efficiency of ER β protein was (93.11±0.57)% (P < 0.05), and after ER β silencing, the expressions of TGF-β1 and BMP-2 were increased by (26.65±3.81)% and (16.62±1.71)% at mRNA level, and increased by (23.79±3.76)% and (18.08±3.20)% at protein level (both P < 0.05). In conclusion, ER β may play an important role in bone metabolism by regulating the expressions of TGF-β1 and BMP-2. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

下腔静脉人工血管置换术后放疗对血管内膜的影响

Objective To investigate the effects of postoperative irradiation on the neointima of artificial blood vessel after prosthetic vessel replacement of inferior vena cava in dogs.Methods Sixteen dogs underwent ePTFE (expended poly-tetra fluoroethylene) prosthetic vessel replacement of inferior vena cava and were then randomly divided into radiotherapy group and control group (n=8 each).For the radiotherapy group, external radiation (35 Gy) was given at two weeks after surgery.Samples were then collected from these two groups on week 8 for study of patency rate of the artificial blood vessel.In addition,HE staining, measurement of neointima thickness, PCNA (proliferating cell nuclear antigen) and CD34 stained immunohistochemistry were performed, and number of endothelial cells (ECs) within 100 Jim neointima were counted.Results The patency rate was 100% (8/8) in radiotherapy group and 75% (6/8)in control group on week 8.The neointima thickness at each segment of artificial blood vessel was significantly decreased in radiotherapy group compared with those in the control group [proximal segment:(610.69±32.90)μm vs(753.39±10.36)μm; mid segment: (530.51 ±32.14 )jun vs(636.55?0.23)μm; distal segment: (544.52±41.99)μm vs (710.39±30.92)μm, all P＜0.01].The percentage counts of PCNA positive cells in each segment of artificial blood vessel were lowered in the radiotherapy group compared with those in the control group [proximal segment: (45.1±7.5)% vs(56.3±7.8)% ; mid segment: (29.2 ±±4.1)% vs(36.6±4.9)%; distal segment: (33.8±5.5)% vs (40.76±.7)%, all P＜0.05].There was no statistical difference in number of ECs within 100 p,m neointima between two groups (P>0.05).Conclusion External radiation (35 Gy) after prosthetic vessel replacement of inferior vena cava has no significant impact on the patency of artificial blood vessel, nor on linings of vascular smooth muscle cells in neointima, but may inhibit the neointima proliferation and PCNA expression.     

结缔组织生长因子诱导人肾小管上皮细胞向间充质细胞转化的实验研究

Objective To investigate the role of connective tissue growth factor ( CTGF) in epithelial mesenchymal transition of HK-2 cells in vitro.Methods HK-2 cells were randomly divided into two groups; (1) control group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum only; and (2) experimental group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum and recombinant CTGF at a final concentration of 5 p-g/L The cells were collected at 72 h time points.Direct immunoiluorescence staining and immunohistochemistry were used to evaluate the E-cadherin,Vimentin,α-SMA and ERK2 in cells.Western-blotting was used to detect the E-cadherin,Vimentin and ERK2 protein expression.Boyden Chamber was used to detect the migration of tubular endothelium at 1 d,3 d and S d.Results There were less E-cadherin but more Vimentin expressed in cells of the experimental group.The presence of α-SMA was detected at 48 h with peak at 72 h in the cells of the experimental group.On the first day,the cellular migration in the two groups showed no difference.However,after 3 days,the transformed cells migrated surpassed the control group with peak at the 5th day [ (45.0±1.1) : (14.0±1.2),P < 0.05 ) ].Conclusion Connective tissue growth factor induces mesenchymal transformation of HK-2 cells,in which the ERK2 signaling pathway may play an important role.     

结缔组织生长因子诱导人肾小管上皮细胞向间充质细胞转化的实验研究

Objective To investigate the role of connective tissue growth factor ( CTGF) in epithelial mesenchymal transition of HK-2 cells in vitro.Methods HK-2 cells were randomly divided into two groups; (1) control group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum only; and (2) experimental group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum and recombinant CTGF at a final concentration of 5 p-g/L The cells were collected at 72 h time points.Direct immunoiluorescence staining and immunohistochemistry were used to evaluate the E-cadherin,Vimentin,α-SMA and ERK2 in cells.Western-blotting was used to detect the E-cadherin,Vimentin and ERK2 protein expression.Boyden Chamber was used to detect the migration of tubular endothelium at 1 d,3 d and S d.Results There were less E-cadherin but more Vimentin expressed in cells of the experimental group.The presence of α-SMA was detected at 48 h with peak at 72 h in the cells of the experimental group.On the first day,the cellular migration in the two groups showed no difference.However,after 3 days,the transformed cells migrated surpassed the control group with peak at the 5th day [ (45.0±1.1) : (14.0±1.2),P < 0.05 ) ].Conclusion Connective tissue growth factor induces mesenchymal transformation of HK-2 cells,in which the ERK2 signaling pathway may play an important role.     

结缔组织生长因子诱导人肾小管上皮细胞向间充质细胞转化的实验研究

Objective To investigate the role of connective tissue growth factor ( CTGF) in epithelial mesenchymal transition of HK-2 cells in vitro.Methods HK-2 cells were randomly divided into two groups; (1) control group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum only; and (2) experimental group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum and recombinant CTGF at a final concentration of 5 p-g/L The cells were collected at 72 h time points.Direct immunoiluorescence staining and immunohistochemistry were used to evaluate the E-cadherin,Vimentin,α-SMA and ERK2 in cells.Western-blotting was used to detect the E-cadherin,Vimentin and ERK2 protein expression.Boyden Chamber was used to detect the migration of tubular endothelium at 1 d,3 d and S d.Results There were less E-cadherin but more Vimentin expressed in cells of the experimental group.The presence of α-SMA was detected at 48 h with peak at 72 h in the cells of the experimental group.On the first day,the cellular migration in the two groups showed no difference.However,after 3 days,the transformed cells migrated surpassed the control group with peak at the 5th day [ (45.0±1.1) : (14.0±1.2),P < 0.05 ) ].Conclusion Connective tissue growth factor induces mesenchymal transformation of HK-2 cells,in which the ERK2 signaling pathway may play an important role.     

结缔组织生长因子诱导人肾小管上皮细胞向间充质细胞转化的实验研究

Objective To investigate the role of connective tissue growth factor ( CTGF) in epithelial mesenchymal transition of HK-2 cells in vitro.Methods HK-2 cells were randomly divided into two groups; (1) control group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum only; and (2) experimental group including cells cultured in DMEM medium supplemented with 10% fetal bovine serum and recombinant CTGF at a final concentration of 5 p-g/L The cells were collected at 72 h time points.Direct immunoiluorescence staining and immunohistochemistry were used to evaluate the E-cadherin,Vimentin,α-SMA and ERK2 in cells.Western-blotting was used to detect the E-cadherin,Vimentin and ERK2 protein expression.Boyden Chamber was used to detect the migration of tubular endothelium at 1 d,3 d and S d.Results There were less E-cadherin but more Vimentin expressed in cells of the experimental group.The presence of α-SMA was detected at 48 h with peak at 72 h in the cells of the experimental group.On the first day,the cellular migration in the two groups showed no difference.However,after 3 days,the transformed cells migrated surpassed the control group with peak at the 5th day [ (45.0±1.1) : (14.0±1.2),P < 0.05 ) ].Conclusion Connective tissue growth factor induces mesenchymal transformation of HK-2 cells,in which the ERK2 signaling pathway may play an important role.