AIM: To establish a model of islet-ductal cell transdifferen-tiation to identify the transdifferentiated cells. METHODS: Collagen was extracted from rat tail at first. Purified rat islets were divided into three group...AIM: To establish a model of islet-ductal cell transdifferen-tiation to identify the transdifferentiated cells. METHODS: Collagen was extracted from rat tail at first. Purified rat islets were divided into three groups, embedded in collagen gel and incubated respectively in DMEM/F12 alone (control group), DMEM/F12 plus epidermal growth factor (EGF), DMEM/F12 plus EGF and cholera toxin (CT). Transdifferentiation was proved by microscopy, RT-PCR, immunohistochemistry and RIA. RESULTS: Islets embedded in collagen gel plus EGF and CT were cystically transformed and could express new gene cytokeratin 19 while still maintaining the expression of insulin and Pdx-1 genes. Immunohistochemistry demonstrated that the protein of cytokeratin 19 was only expressed in the third group. The insulin content secreted by islets in the third group decreased significantly during the transdiffe-rentiation. CONCLUSION: CT is a crucial factor for the islet-ductal cell transdifferentiation.展开更多
目的:探讨苯妥英钠(PHT)对血管内皮细胞转分化作用的影响及其作用机制,为提高牙周组织再生能力的治疗提供思路。方法:通过transwell建立RAOEC与大鼠巨噬细胞共培养体系。研究设为空白组:基础培养基;P0组:基础培养基+100 ng/m L LPS;P25...目的:探讨苯妥英钠(PHT)对血管内皮细胞转分化作用的影响及其作用机制,为提高牙周组织再生能力的治疗提供思路。方法:通过transwell建立RAOEC与大鼠巨噬细胞共培养体系。研究设为空白组:基础培养基;P0组:基础培养基+100 ng/m L LPS;P25组:基础培养基+100 ng/m L LPS+25μg/m L PHT;P50组:基础培养基+100 ng/m L LPS+50μg/m L PHT,每组设3个复孔。T组:利用8 cm2培养皿单独接种RAOEC细胞,添加含有10μg/m L TGF-β1的培养基培养。Elisa检测不同处理组中TGF-β1的分泌量。RT-PCR及Western-blot,鉴定RAOEC转分化的标志CD31、α-SMA、SCF的mRNA及蛋白表达量。结果:一定浓度范围内,随着PHT浓度的升高,大鼠血管内皮细胞表型发生明显转变;ELISA检测结果显示,P0组、P25组、P50组细胞上清液TGF-β1含量分别为1.87±1.01、5.33±0.94、8.96±1.25,差异具有统计学意义(P<0.05);RT-PCR检测结果显示,P0、P25、P50组间比较,CD31 mRNA表达量逐步降低,α-SMA与SCF mRNA表达量逐步升高,差异具有统计学意义(P<0.05);Western-blot灰度分析显示,P0、P25、P50组间比较,CD31灰度值逐步降低,α-SMA与SCF灰度值逐步增高,差异具有统计学意义(P<0.05)。结论:PHT具有促进大鼠血管内皮细胞转分化的作用,为PHT应用于牙周组织再生提供了新思路。展开更多
基金Supported by the National Natural Science Foundation of China, No. 30200136
文摘AIM: To establish a model of islet-ductal cell transdifferen-tiation to identify the transdifferentiated cells. METHODS: Collagen was extracted from rat tail at first. Purified rat islets were divided into three groups, embedded in collagen gel and incubated respectively in DMEM/F12 alone (control group), DMEM/F12 plus epidermal growth factor (EGF), DMEM/F12 plus EGF and cholera toxin (CT). Transdifferentiation was proved by microscopy, RT-PCR, immunohistochemistry and RIA. RESULTS: Islets embedded in collagen gel plus EGF and CT were cystically transformed and could express new gene cytokeratin 19 while still maintaining the expression of insulin and Pdx-1 genes. Immunohistochemistry demonstrated that the protein of cytokeratin 19 was only expressed in the third group. The insulin content secreted by islets in the third group decreased significantly during the transdiffe-rentiation. CONCLUSION: CT is a crucial factor for the islet-ductal cell transdifferentiation.
文摘目的:探讨苯妥英钠(PHT)对血管内皮细胞转分化作用的影响及其作用机制,为提高牙周组织再生能力的治疗提供思路。方法:通过transwell建立RAOEC与大鼠巨噬细胞共培养体系。研究设为空白组:基础培养基;P0组:基础培养基+100 ng/m L LPS;P25组:基础培养基+100 ng/m L LPS+25μg/m L PHT;P50组:基础培养基+100 ng/m L LPS+50μg/m L PHT,每组设3个复孔。T组:利用8 cm2培养皿单独接种RAOEC细胞,添加含有10μg/m L TGF-β1的培养基培养。Elisa检测不同处理组中TGF-β1的分泌量。RT-PCR及Western-blot,鉴定RAOEC转分化的标志CD31、α-SMA、SCF的mRNA及蛋白表达量。结果:一定浓度范围内,随着PHT浓度的升高,大鼠血管内皮细胞表型发生明显转变;ELISA检测结果显示,P0组、P25组、P50组细胞上清液TGF-β1含量分别为1.87±1.01、5.33±0.94、8.96±1.25,差异具有统计学意义(P<0.05);RT-PCR检测结果显示,P0、P25、P50组间比较,CD31 mRNA表达量逐步降低,α-SMA与SCF mRNA表达量逐步升高,差异具有统计学意义(P<0.05);Western-blot灰度分析显示,P0、P25、P50组间比较,CD31灰度值逐步降低,α-SMA与SCF灰度值逐步增高,差异具有统计学意义(P<0.05)。结论:PHT具有促进大鼠血管内皮细胞转分化的作用,为PHT应用于牙周组织再生提供了新思路。
