Objective To investigate the effect of transforming growth factor-β1 (TGF-β1) gene transfer on the biological characteristics of osteoblasts. Methods The expression of TGF-β1 in the transfected osteoblasts was detected by in situ hybridization and assay of TGF-β1 activity in the supernatant (minklung epithelium cell growth -inhibition test). The effects of gene transfer andsupernatant of the transfected osteoblasts on the proliferation and alkaline phosphatase(ALP) activity of osteoblasts were detected by 3 H-TdR and MTT. Results The results of in situ hybridization analysis suggested that the osteoblasts transfected by TGF-β1 gene could express TGF-β1 obviously. The complex medium, which was the mixture of serum-free DMEM and the activated supernatant according to 1∶1, 1∶2, 1∶4, could inhibit growth of Mv-1-Lu evidently and the ratios ofinhibition were 16.3%, 22.7%, 28.2% respectively. TGF-β1 gene transfer hadno effect on the biological characteristics of osteoblasts, but the activated supernatant of transfected osteoblasts stimulated proliferation and inhibited ALPactivity of osteoblasts. Conclusion TGF-β1 gene transfer promotes the expression of TGF-β1 and the biological characteristics of trasfected osteoblasts are stable, which is helpful for gene therapy of bone defects in vivo.
This study is aimed to investigate the effects of mechanical stretch on the expression of transforming growth factor-β1 (TGF-β1) and fibroblast growth factor-2 (FGF-2), and the signaling pathway in human bronchial epithelioid (16HBE) cells under mechanical stretch. Using loading device with flexible substrate (FX-4000T) to stretch 16HBE cells, we found that the stretching elongation was 15%, at frequency of 1 Hz, stretching for 0.5 h, 1 h, 1.5 h and 2 h. Choosing the higher expression of TGF-β1, FGF-2 and Ca2+ group to carry out intervention experiments, we used the cells pretreated with canonical transient receptor potential 1 (TRPC1) channel antagonist SKF96365, protein kinase C (PKC) inhibitor HA-100, and thereafter mechanical stretch to interpose. Compared with those in the blank control group, TGF-β1 and FGF-2' protein and mRNA, intracellular Ca2+ fluorescence intensity were higher, and the differences were statistically significant (P < 0.05) at the 4 time points, 0.5 h, 1 h, 1.5 h and 2 h. At 0.5 h, the increasing rate was the highest. TGF-β1 protein and mRNA, FGF-2 protein and mRNA, intracellular Ca2+ luorescence intensity in the stretch+SKF96365 and stretch+HA-100 intervented group were decreased, the differences were statistically significant than those in 0.5 h stretch group (P < 0.05) without intervention. The expression of TGF-β1, FGF-2 was up-regulated in 16HBE cells under mechanical stretch, PKC, TRPC1, and Ca2+ may participate in the signal path.
【Abstract】Objective To investigate the expression of tumor growth tactor β1 (TGFβ1) and p27 in gallbladder carcinoma and their relation to the development of the carcinoma. Methods The expression of TGF-β1 and p27 in 36 cases of gallbladder carcinoma was detected by SP immunohistochemical staining. Twenty cases of chronic cholecystitis were collected as control. Results The positive rate of TGF-β1 （63.9%） was higher than that of the control （10.0%），P＜0.05， and the positive rate of p27 (47.2%) was lower than that of the control 100%(P＜0.05). The positive rate of TGF-β1 was significantly higher in metastasis or Nevin Ⅳ～Ⅴ stage cases than that of non-metastasis or Nevin Ⅰ～Ⅲ stage cases 33.3% (P＜0.05). The positive rate of p27 was statistically higher in moderate and highly differentiation (60.9%), nonmetastasis (75.0%) or Nevin’s Ⅰ～Ⅲ stage (75.0%) cases than those of poor differentiation (23.0%), metastasis (33.3%) and Nevin Ⅳ～Ⅴ stage (33.3%) cases (P＜0.05). The expression of p27 was negatively correlated with that of TGF-β1(r=-0.4473，P＜0.05). There was significant difference in survival time between patients with TGF-β1 positive and TGF-β1 negative（P＜0.05）. The difference was also found between patients with p27 positive and p27 negative. Conclusion The upregulation of TGF-β1 and downregulation of p27 in gallbladder carcinoma indicates the imbalance of TGF-β1/p27 system, which may play a role in the carcinogenesis and predict the malignant behaviors of the carcinoma.
Objective To investigate the effect of imatinib mesylate on radiation-induced lung injury mice and its influence on the oxidative stress and transforming growth factor-β1 (TGF-β1) expression in mice. Methods Forty-five C57BL/6 mice were divided into a treatment group, a control group and a model group. The treatment group and model group were given radiation of 18 Gy delivered in the thorax. After 4 h daily of the radiation, the treatment group received imatinib mesylate of 0.081 g/kg, while the other groups were given normal saline solution. The experiments were continued for 30 days. After the experiments, the lungs of mice were divided into 4 parts. The haematoxylin and eosin and immunohistochemical stain were prepared to observe the situation of pathology and TGF-β1. The lung homogenate was prepared and the levels of superoxide dismutase (SOD), malondialdehyde (MDA), total antioxidant capacity (T-Aoc) and glutathione peroxidase (GSH-PX) were detected. Results The levels of GSH-PX, T-Aoc and SOD were (173.15±12.21) U, (119.33±11.06) U/mgprot and (1.73±0.33) nmol/mgprot in the treatment group, significantly higher than the control group, while the levels of MDA was (0.68±0.08) nmol/mgprot, significantly lower than the control group (P<0.05). The HE and immunohistochemical stain showed that there were mild alveolar inflammatory changes in the treatment group while such changes were serious in the model group. The scores of HE and immunohistochemical were 1.26±0.12 and 0.31±0.12 in the treatment group, significantly lower than those in the control group (P<0.05). Conclusion The imatinib mesylate can effectively ameliorate the oxidative stress and inhibite TGF-β1 expression in radiation-induced lung injury mice.
Objective To study the influence of transforming growth factor-β1(TGF-β1), dentin non-collagen proteins(dNCPs) and their complexon tissue engineering pulp system. Methods Collagen I and dentin powder were used to construct the system of pulp cells in 3dimensional culture, dentin powder was added in the gel. The tissue engineering pulp were divided TGF-β1 group, dNCPs group, TGF-β1/dNCPsgroup and control group.After3, 6 and 14 days, the appearance and the differentiation of pulp cells were observed by HE staining and immunohistochemical staining -respectively. Results Collagen I could form netted collagen gel construction. Growing condition of pulp cells in gel was similar to that of pulp cells in vivo. After the TGF-β1 and dNCPswere added, the pulp cells had some characteristics of odontoblasts and had unilateral cell process after culture 6 days. Pulp cells arranged with parallel columnar and form dentin-pulp-like complex after 14 days. Immunohistochemical staining showed dentin salivary protein(DSP) began to express in some cells.The number of positive cell was most in the TGF-β1 group. No positive cells were detected in the control group. Conclusion The transforming growth factor-β1 and noncollagen proteins can stimulate the pulp cells to transform into odontoblasts to some extent, which promote the formation of tissue engineering pulp.
ObjectiveTo investigate the early diagnostic value of transforming growth factor-β1(TGF-β1) on acute rejection after liver transplantation in rhesus by detecting the expression of TGF-β1 in the liver tissue. MethodsLiver transplantation models in rhesus were constructed by the improved vascular dual cuff, supporting tube of biliary tract, and artery anastomosis method.The successful models were randomly divided into experimental group (no immunosuppressant treatment in perioperative period) and control group (treated by immunosuppressant in perioperative period).Then the blood samples and liver tissues were collected at 6, 12, 24, and 72 hours after surgery.Allograft rejections of liver tissue after liver transplantation were monitored by liver function test, hematoxylin-eosin staining and Banff score.Finally, the expression level of TGF-β1 was detected by Western blot analysis or immunohistochemistry technique. Results①The acute rejection happened in all the rhesus at 12 h, 24 h and 72 h after liver transplantation, especially at 72 h after liver transplantation in the experimental group, the Banff grade levels of acute rejection in the liver tissue was more severe than that in the control group (P < 0.05).②The levels of ALT, AST, and TBIL after liver transplantation was gradually increased, which were similar at 6 h and 12 h after transplantation between the two groups, but which at 24 h and 72 h after transplantation in the experimental group were significantly higher than those in the control group (P < 0.05).③The results of TGF-β1 protein expression using immunohistochemical detection:The percentage of positive area of TGF-β1 of liver tissue at 12 h in the experimental group was significantly higher than that in the control group (P < 0.05).With the extension of time, it was gradually increased and significantly higher than that in the control group at 24 h or 72 h (P < 0.05).④The semi-quantitative results of TGF-β1 protein expression using Western blot detection:The TGF-β1 protein expressions began to increase at 6 h after liver transplantation in the experimental group and the control group, and the magnitude of increase was more obvious in the experimental group.The TGF-β1 protein expressions at different time (6 h, 12 h, 24 h, and 72 h) in the experimental group were significantly higher than those in the control group (P value was 0.003, 0.001, 0.001, and 0.001, respectively). ConclusionsThe elevated level of TGF-β1 of liver tissue after liver transplantation might suggest the enhanced cellular immune function, it might have certain significance for early diagnosis of acute rejection after liver transplantation.
Long non-coding RNA (lncRNA) Dnm3os plays a critical role in peritendinous fibrosis and pulmonary fibrosis, but its role in the process of cardiac fibrosis is still unclear. Therefore, we carried out study by using the myocardial fibrotic tissues obtained by thoracic aortic constriction (TAC) in an early study of our group, and the in vitro cardiac fibroblast activation model induced by transforming growth factor-β1 (TGF-β1). Quantitative real-time polymerase chain reaction (RT-qPCR), Western blot, and collagen gel contraction test were used to identify the changes of activation phenotype and the expression of Dnm3os in cardiac fibroblasts. Small interfering RNA was used to silence Dnm3os to explore its role in the activation of cardiac fibroblasts. The results showed that the expression of Dnm3os was increased significantly in myocardial fibrotic tissues and in the activated cardiac fibroblasts. And the activation of cardiac fibroblasts could be alleviated by Dnm3os silencing. Furthermore, the TGF-β1/Smad2/3 pathway was activated during the process of cardiac fibroblasts activation, while was inhibited after silencing Dnm3os. The results suggest that Dnm3os silencing may affect the process of cardiac fibroblast activation by inhibiting TGF-β1/Smad2/3 signal pathway. Therefore, interfering with the expression of lncRNA Dnm3os may be a potential target for the treatment of cardiac fibrosis.
目的 研究胃癌细胞SGC-7901培养上清液及转化生长因子-β1（TGF-β1）是否可促进人类腹膜间皮细胞表达βig-h3蛋白。方法 培养胃癌细胞SGC-7901，取第3天培养液上清与DMEM培养液的混合液 （1∶4）以及0、1.0、10.0和50.0ng/ml的 TGF-β1分别刺激人类腹膜间皮细胞HMrSV50、3、6、12及24h，ELISA方法检测上清液中βig-h3蛋白浓度，Western blot法检测细胞内βig-h3蛋白浓度。结果 对照组有基础量的βig-h3蛋白表达； 胃癌细胞SGC-7901培养上清液及TGF-β1均可明显增加HMrSV5细胞上清液及细胞内的βig-h3蛋白浓度（P＜0.05），且TGF-β1的刺激作用呈时间及浓度依赖性。结论 胃癌细胞SGC-7901培养上清液及TGF-β1可明显刺激HMrSV5细胞表达和分泌βig-h3蛋白。
OBJECTIVE: To explore the molecular mechanisms involved in the increased collagen synthesis by platelet-derived wound healing factors (PDWHF) during wound healing in alloxan-induced diabetic rats. METHODS: Thirty-three male SD rats were divided into two groups, the normal (n = 9) (group A) and the diabetic group (n = 24). Two pieces of full-thickness skin with diameter of 1.8 cm were removed from the dorsal site of diabetic rats. PDWHF (100 micrograms/wound) was topically applied to one side of the diabetic wounds (group B) on the operation day and then once a day in the next successive 6 days. Meanwhile, bovine serum albumin (100 micrograms/wound) was applied to the other side of diabetic wound as control group (group C) in the same way. Levels of transforming growth factor-beta 1 (TGF-beta 1) and procollagen I mRNA in wound tissue were inspected by dot blotting. RESULTS: TGF-beta 1 mRNA levels in group B were 4 folds and 5.6 folds compared with those in group C after 5 and 7 days (P lt; 0.01), however, still significantly lower than those of group A (P lt; 0.05). There was no significance difference among three groups on the 10th day after wounding. The levels for procollagen I mRNA in group B amounted to 2.1, 1.8 and 2.3 folds of those in group C after 5, 7, and 10 days (P lt; 0.01), respectively. Compared with those in the group A, procollagen I mRNA levels in the group B were significantly lower after 5 and 7 days (P lt; 0.05), and no significant difference was observed between group B and A after 10 days. CONCLUSION: One important way for PDWHF to enhance the collagen synthesis in diabetic wound healing is to increase the gene expression of endogenous TGF-beta 1.
Objective To investigate the role of IFN-γ in suppressing bleomycin-induced pulmonary fibrosis in rats.Methods Seventy-five SD rats were randomly divided into five groups (15 rats in each group)，ie.a normal group，a bleomycin-induced pulmonary fibrosis model group，a dexamethasone-treated group，a high-dose IFN-γ-treated group (150 000 U/kg) and a low-dose IFN-γ-treated group (50 000 U/kg).Five rats in each group were randomly killed in 7th day，14th day and 28th day after relative treatment respectively，and lung tissue samples were harvested for histopathology study.HE and Masson staining were used to determine the extent of alveolus inflammation and pulmonary fibrosis respectively.Histoimmunochemical method were adapted to determine protein levels of TGF-β1，CTGF，type Ⅰcollagen and type Ⅲ collagen in pulmonary tissues.Results Histopathological study showed that treatment with either dexamethasone or IFN-γ (both high dose and low dose) remarkably meliorated the extent of alveolus inflammation and suppressed pulmonary fibrosis (compared with model group，all Plt;0.05).Histoimmunochemical study suggested that both dexamethasone and IFN-γ could inhibit the expression of TGF-β1，CTGF，type Ⅰand type Ⅲ collagen (compared with model group，all Plt;0.05)，and the suppression of TGF-β1，type Ⅰand type Ⅲ collagen expression was more obvious in high-dose IFN-γ-treated group than those in low-dose group (Plt;0.05).Conclusions INF-γ possesses apparent anti-fibrosis effect that is similar to dexamethasone but with less side effect.Such effect may resulted from reduced production of type Ⅰand type Ⅲ collagen through expression inhibition of cytokines such as TGF-β1 and CTGF.