AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells(BMSCs) with human urokinase-type plasminogen activator(u PA) on liver fibrosis, and to investigate the mechanism of gene therapy.M...AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells(BMSCs) with human urokinase-type plasminogen activator(u PA) on liver fibrosis, and to investigate the mechanism of gene therapy.METHODS: BMSCs transfected with adenovirusmediated human urokinase plasminogen activator(Adu PA) were transplanted into rats with CCl4-induced liver fibrosis. All rats were sacrificed after 8 wk, and their serum and liver tissue were collected for biochemical, histopathologic, and molecular analyzes. The degree of liver fibrosis was assessed by hematoxylin and eosin or Masson's staining. Western blot and quantitative reverse transcription-polymerase chain reaction were used to determine protein and m RNA expression levels.RESULTS: Serum levels of alanine aminotransferase, aminotransferase, total bilirubin, hyaluronic acid, laminin, and procollagen type Ⅲ were markedly decreased, whereas the levels of serum albumin were increased by u PA gene modified BMSCs treatment. Histopathology revealed that chronic CCl4-treatment resulted in significant fibrosis while u PA gene modified BMSCs treatment significantly reversed fibrosis. By quantitatively analysing the fibrosis area of liver tissue using Masson staining in different groups of animals, we found that model animals with CCl4-induced liver fibrosis had the largest fibrotic area(16.69% ± 1.30%), while fibrotic area was significantly decreased by BMSCs treatment(12.38% ± 2.27%) and was further reduced by u PA-BMSCs treatment(8.31% ± 1.21%). Both protein and m RNA expression of β-catenin, Wnt4 and Wnt5 a was down-regulated in liver tissues following u PA gene modified BMSCs treatment when compared with the model animals.CONCLUSION: Transplantation of u PA gene modified BMSCs suppressed liver fibrosis and ameliorated liver function and may be a new approach to treating liver fibrosis. Furthermore, treatment with u PA gene modified BMSCs also resulted in a decrease in expression of molecules of the Wnt signaling pathway.展开更多
基金江苏省2015年度普通高校研究生科研创新计划项目(No KYZZ15-0274)The People Programme(Marie Curie Actions)of the European Union’s Seventh Framework Programme FP7/2007-2013/under REA grant agreement n°PIR SES-GA-2013-612589)+1 种基金江苏省优势学科和江苏省自然科学基金项目(No BK20131415)国家自然科学基金资助项目(No 81503374)
基金Supported by National Natural Science Foundation of ChinaNo.81460114+5 种基金Natural Science Foundation of Guangxi Zhuang Autonomous RegionNo.1355005-3-2 and No.2012GXNSFAA053143Chinese Traditional Medicine Science Foundation of Guangxi Zhuang Autonomous RegionNo.GZPT1238Science Foundation of Guangxi Department of EducationNo.201203YB036 and No.2013LX031
文摘AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells(BMSCs) with human urokinase-type plasminogen activator(u PA) on liver fibrosis, and to investigate the mechanism of gene therapy.METHODS: BMSCs transfected with adenovirusmediated human urokinase plasminogen activator(Adu PA) were transplanted into rats with CCl4-induced liver fibrosis. All rats were sacrificed after 8 wk, and their serum and liver tissue were collected for biochemical, histopathologic, and molecular analyzes. The degree of liver fibrosis was assessed by hematoxylin and eosin or Masson's staining. Western blot and quantitative reverse transcription-polymerase chain reaction were used to determine protein and m RNA expression levels.RESULTS: Serum levels of alanine aminotransferase, aminotransferase, total bilirubin, hyaluronic acid, laminin, and procollagen type Ⅲ were markedly decreased, whereas the levels of serum albumin were increased by u PA gene modified BMSCs treatment. Histopathology revealed that chronic CCl4-treatment resulted in significant fibrosis while u PA gene modified BMSCs treatment significantly reversed fibrosis. By quantitatively analysing the fibrosis area of liver tissue using Masson staining in different groups of animals, we found that model animals with CCl4-induced liver fibrosis had the largest fibrotic area(16.69% ± 1.30%), while fibrotic area was significantly decreased by BMSCs treatment(12.38% ± 2.27%) and was further reduced by u PA-BMSCs treatment(8.31% ± 1.21%). Both protein and m RNA expression of β-catenin, Wnt4 and Wnt5 a was down-regulated in liver tissues following u PA gene modified BMSCs treatment when compared with the model animals.CONCLUSION: Transplantation of u PA gene modified BMSCs suppressed liver fibrosis and ameliorated liver function and may be a new approach to treating liver fibrosis. Furthermore, treatment with u PA gene modified BMSCs also resulted in a decrease in expression of molecules of the Wnt signaling pathway.
文摘目的探讨在体外环境下芝麻素对小鼠骨髓间充质干细胞向成骨细胞分化的影响。方法从大鼠股骨中提取骨髓间充质干细胞(bone marrow stromal cells,BMSCs),诱导其向成骨细胞分化。使用FH535(1μmol/L)沉默Wnt/β-catenin信号通路,建立沉默组,同时建立非沉默组。两组同时给予芝麻素(0、1或10μmol/L)干预,检测细胞增殖/毒性,同时检查碱性磷酸酶(alkaline phosphatase,ALP)、成骨相关转录因子抗体(osterix,OSX)、SRY-box 9(SOX9)、Runt相关转录因子2(recombinant runt related transcription factor 2,RUNX2)、骨钙素(osteocalcin,OCN)、b-连环蛋白(β-catenin)、低密度脂蛋白受体相关蛋白5(low density lipoprotein receptor-related protein 5,LRP5)以及糖原合成酶激酶-3b(glycogen synthase kinase-3b,gsk-3b)的表达水平。结果芝麻素对BMSCs的增殖无明显影响,且无细胞毒性作用。较高浓度的芝麻素诱导Wnt/β-catenin信号通路,增加ALP、OSX、SOX9、RUNX2和OCN的表达(P<0.05)。沉默Wnt/β-catenin后,RUNX2和OCN表达减弱。结论芝麻素有通过调节Wnt/β-catenin信号通路促进大鼠骨髓间充质干细胞向成骨细胞分化的趋势,对重塑大鼠骨结构有一定影响。芝麻素对骨质疏松有治疗和预防作用。
