Objective To explore the effect of TGF-a1 on the mRNA and prot ei n expression of connective tissue growth factor (CTGF) in human peritoneal mesot helial cells (HPMCs) and the probable mechanism. Methods Greater oment...Objective To explore the effect of TGF-a1 on the mRNA and prot ei n expression of connective tissue growth factor (CTGF) in human peritoneal mesot helial cells (HPMCs) and the probable mechanism. Methods Greater omenta from hea lthy adults were used for primary culture of HPMCs. The third generation cells w ere stimulated by 5 ng/ml TGF-a1. Immunohistochemistry, Western blot, ELISA an d RT-PCR were employed to examine the follows:the mRNA and protein expression o f CTGF, the mRNA and protein expression of fibronectin (FN) and collagen type Ⅰ (ColⅠ), the protein expression of p-Smad2/3 and its migration in HPMCs. Result s (1)The mRNA expression of CTGF in the treatment groups increased obviously a s compared to the control group, and peaked at 48 h. (2)The protein expression of intracellular CTGF in the control group was very little, whereas it increase d markedly 24 h after TGF-a1 stimulation, and peaked at 48 h. The mRNA express ion of intracellular FN, and the protein expression of supernatent FN, as well a s the mRNA and protein expression of intracellular ColⅠin the treatment group u p-regulated significantly in a time dependent manner as compared to those in th e control group. (3)p-Smad2/3 in HPMCs hardly expressed in the control group( 3%p-Smad2/3-positive cells), and remarkably expressed 15 min after TGF-a1 s timulation(29%), peaked at 1 h (84%) and then decreased after 2 h(37%). Meanw hile, p-Smad2/3 mainly distributed in cytoplasm at 15 min, concentrated in cell nucleus and peri-nucleus at 1 h, and distributed in cytoplasm again at 2 h. Co nclusions TGF-a1 can induce the mRNA and protein expression of CTGF in the pro gression of human peritoneal fibrosis. The probably mechanism is that signaling through Smad pathway in HPMCs can be activated by TGF-a1 stimulation.展开更多
转化生长因子-β(transform ing growth factor-,βTGF-β)是一类多功能多肽类生长因子,对细胞增殖分化、细胞外基质产生、血管生成、细胞凋亡及机体免疫系统均起着非常重要的调节作用。体内外实验证实TGF-β及其受体的表达均受到低氧...转化生长因子-β(transform ing growth factor-,βTGF-β)是一类多功能多肽类生长因子,对细胞增殖分化、细胞外基质产生、血管生成、细胞凋亡及机体免疫系统均起着非常重要的调节作用。体内外实验证实TGF-β及其受体的表达均受到低氧的调节,但这种调节具有明显的组织细胞特异性,而且不同亚型的TGF-β及不同类型的受体对低氧的反应是通过不同的信号分子或转录因子来介导的。展开更多
目的:探讨转化生长因子-β_1诱导的大鼠心肌细胞肥大中ERK信号通路的蛋白表达。方法:建立TGF-β_1诱导的体外大鼠心肌细胞肥大模型。PI染色标记细胞双链RNA法检测心肌细胞RNA表达量,间接检测心肌细胞肥大。实时荧光定量PCR(Real time P...目的:探讨转化生长因子-β_1诱导的大鼠心肌细胞肥大中ERK信号通路的蛋白表达。方法:建立TGF-β_1诱导的体外大鼠心肌细胞肥大模型。PI染色标记细胞双链RNA法检测心肌细胞RNA表达量,间接检测心肌细胞肥大。实时荧光定量PCR(Real time PCR)检测心肌细胞肥大相关基因肌球蛋白重链β亚型(β-MHC)的表达。Western-blot检测心肌细胞中ERK1/2蛋白表达。结果:TGF-β_1可诱导体外培养心肌细胞肥大基因的表达,β-MHC的表达明显高于对照组(P<0.01)。TGF-β_1组PI含量明显增高(P<0.01),与对照组相比,TGF-β_1可明显上调ERK通路中磷酸化ERK1/2蛋白表达增加。RNA干扰技术抑制TGF-β_1的特异性通路Smad通路中Smad2蛋白表达后,磷酸化ERK1/2表达也被部分抑制(P<0.01)。结论:TGF-β_1可诱导心肌细胞肥大的形成,同时ERK通路中ERK1/2表达增加,TGF/Smad通路与ERK通路中ERK1/2可能存在交互作用。展开更多
文摘Objective To explore the effect of TGF-a1 on the mRNA and prot ei n expression of connective tissue growth factor (CTGF) in human peritoneal mesot helial cells (HPMCs) and the probable mechanism. Methods Greater omenta from hea lthy adults were used for primary culture of HPMCs. The third generation cells w ere stimulated by 5 ng/ml TGF-a1. Immunohistochemistry, Western blot, ELISA an d RT-PCR were employed to examine the follows:the mRNA and protein expression o f CTGF, the mRNA and protein expression of fibronectin (FN) and collagen type Ⅰ (ColⅠ), the protein expression of p-Smad2/3 and its migration in HPMCs. Result s (1)The mRNA expression of CTGF in the treatment groups increased obviously a s compared to the control group, and peaked at 48 h. (2)The protein expression of intracellular CTGF in the control group was very little, whereas it increase d markedly 24 h after TGF-a1 stimulation, and peaked at 48 h. The mRNA express ion of intracellular FN, and the protein expression of supernatent FN, as well a s the mRNA and protein expression of intracellular ColⅠin the treatment group u p-regulated significantly in a time dependent manner as compared to those in th e control group. (3)p-Smad2/3 in HPMCs hardly expressed in the control group( 3%p-Smad2/3-positive cells), and remarkably expressed 15 min after TGF-a1 s timulation(29%), peaked at 1 h (84%) and then decreased after 2 h(37%). Meanw hile, p-Smad2/3 mainly distributed in cytoplasm at 15 min, concentrated in cell nucleus and peri-nucleus at 1 h, and distributed in cytoplasm again at 2 h. Co nclusions TGF-a1 can induce the mRNA and protein expression of CTGF in the pro gression of human peritoneal fibrosis. The probably mechanism is that signaling through Smad pathway in HPMCs can be activated by TGF-a1 stimulation.
文摘转化生长因子-β(transform ing growth factor-,βTGF-β)是一类多功能多肽类生长因子,对细胞增殖分化、细胞外基质产生、血管生成、细胞凋亡及机体免疫系统均起着非常重要的调节作用。体内外实验证实TGF-β及其受体的表达均受到低氧的调节,但这种调节具有明显的组织细胞特异性,而且不同亚型的TGF-β及不同类型的受体对低氧的反应是通过不同的信号分子或转录因子来介导的。
文摘目的:探讨转化生长因子-β_1诱导的大鼠心肌细胞肥大中ERK信号通路的蛋白表达。方法:建立TGF-β_1诱导的体外大鼠心肌细胞肥大模型。PI染色标记细胞双链RNA法检测心肌细胞RNA表达量,间接检测心肌细胞肥大。实时荧光定量PCR(Real time PCR)检测心肌细胞肥大相关基因肌球蛋白重链β亚型(β-MHC)的表达。Western-blot检测心肌细胞中ERK1/2蛋白表达。结果:TGF-β_1可诱导体外培养心肌细胞肥大基因的表达,β-MHC的表达明显高于对照组(P<0.01)。TGF-β_1组PI含量明显增高(P<0.01),与对照组相比,TGF-β_1可明显上调ERK通路中磷酸化ERK1/2蛋白表达增加。RNA干扰技术抑制TGF-β_1的特异性通路Smad通路中Smad2蛋白表达后,磷酸化ERK1/2表达也被部分抑制(P<0.01)。结论:TGF-β_1可诱导心肌细胞肥大的形成,同时ERK通路中ERK1/2表达增加,TGF/Smad通路与ERK通路中ERK1/2可能存在交互作用。
