The relation between the expression and activity of MMP-9 in C-reactive protein （CRP）-induced human THP-1 mononuclear cells and the activation of nuclear factor kappa-B （NF-κB） was studied to investigate the poss...The relation between the expression and activity of MMP-9 in C-reactive protein （CRP）-induced human THP-1 mononuclear cells and the activation of nuclear factor kappa-B （NF-κB） was studied to investigate the possible role of CRP in plaque destabilization. Human THP-1 cells were incubated in the presence of CRP at 0 （control group）, 25, 50 and 100 μg/mL （CRP groups） for 24 h. In PDTC （a specific NF-κB inhibitor） group, the cells were pre-treated with PDTC at 10 μmol/L and then with 100 μg/mL CRP. The conditioned media （CM） and human THP-1 cells in different groups were harvested. MMP-9 expression in CM and human THP-1 cells was measured by ELISA and Western blotting. MMP-9 activity was assessed by fluorogenic substrates. The expression of NF-κB inhibitor α （IκB-α） and NF-κB p65 was detected by Western blotting and ELISA respectively. The results showed that CRP increased the expression and activity of MMP-9 in a dose-dependent manner in the human THP-1 cells. Western blotting revealed that IiB-α expression was decreased in the cells with the concentrations of CRP and ELISA demonstrated that NF-κB p65 expression in the CRP-induced cells was increased. After pre-treatment of the cells with PDTC at 10 μmol/L, the decrease in IκB-α expression and the increase in NF-κB p65 expression in the CRP-induced cells were inhibited, and the expression and activity of MMP-9 were lowered too. It is concluded that increased expression and activity of MMP-9 in CRP-induced human THP-1 cells may be associated with activation of NF-κB. Down-regulation of the expression and activity of MMP-9 may be a new treatment alternative for plaque stabilization by inhibiting the NF-κB activation.展开更多
目的:探讨围产期缺血缺氧对新生大鼠肺超微结构和肿瘤坏死因子α(TNFα-)及核因子κB抑制蛋白(κI-Bα)表达的影响。方法:结扎孕鼠一侧子宫角血管(另一侧宫内胎鼠作为对照),建立围产期缺血缺氧动物模型。电镜下观察肺组织超微结构变化...目的:探讨围产期缺血缺氧对新生大鼠肺超微结构和肿瘤坏死因子α(TNFα-)及核因子κB抑制蛋白(κI-Bα)表达的影响。方法:结扎孕鼠一侧子宫角血管(另一侧宫内胎鼠作为对照),建立围产期缺血缺氧动物模型。电镜下观察肺组织超微结构变化并分别采用ELISA、RT-PCR和W estern b lot杂交法观察不同程度缺血缺氧新生大鼠肺组织TNFα-蛋白和mRNA及I-κBα表达的变化。结果:宫内急性缺血缺氧20 m in时,新生大鼠Ⅱ型肺泡上皮细胞和毛细血管内皮细胞受损;肺组织TNFα-蛋白和mRNA表达明显增强(P均<0.01),κI-Bα表达明显减弱(P<0.05),并随缺血缺氧时间延长病变加重。结论:围产期急性缺血缺氧致新生大鼠肺损伤,Iκ-Bα和TNFα-的异常表达可能是肺损伤的重要原因。展开更多
文摘The relation between the expression and activity of MMP-9 in C-reactive protein （CRP）-induced human THP-1 mononuclear cells and the activation of nuclear factor kappa-B （NF-κB） was studied to investigate the possible role of CRP in plaque destabilization. Human THP-1 cells were incubated in the presence of CRP at 0 （control group）, 25, 50 and 100 μg/mL （CRP groups） for 24 h. In PDTC （a specific NF-κB inhibitor） group, the cells were pre-treated with PDTC at 10 μmol/L and then with 100 μg/mL CRP. The conditioned media （CM） and human THP-1 cells in different groups were harvested. MMP-9 expression in CM and human THP-1 cells was measured by ELISA and Western blotting. MMP-9 activity was assessed by fluorogenic substrates. The expression of NF-κB inhibitor α （IκB-α） and NF-κB p65 was detected by Western blotting and ELISA respectively. The results showed that CRP increased the expression and activity of MMP-9 in a dose-dependent manner in the human THP-1 cells. Western blotting revealed that IiB-α expression was decreased in the cells with the concentrations of CRP and ELISA demonstrated that NF-κB p65 expression in the CRP-induced cells was increased. After pre-treatment of the cells with PDTC at 10 μmol/L, the decrease in IκB-α expression and the increase in NF-κB p65 expression in the CRP-induced cells were inhibited, and the expression and activity of MMP-9 were lowered too. It is concluded that increased expression and activity of MMP-9 in CRP-induced human THP-1 cells may be associated with activation of NF-κB. Down-regulation of the expression and activity of MMP-9 may be a new treatment alternative for plaque stabilization by inhibiting the NF-κB activation.
文摘目的:探讨围产期缺血缺氧对新生大鼠肺超微结构和肿瘤坏死因子α(TNFα-)及核因子κB抑制蛋白(κI-Bα)表达的影响。方法:结扎孕鼠一侧子宫角血管(另一侧宫内胎鼠作为对照),建立围产期缺血缺氧动物模型。电镜下观察肺组织超微结构变化并分别采用ELISA、RT-PCR和W estern b lot杂交法观察不同程度缺血缺氧新生大鼠肺组织TNFα-蛋白和mRNA及I-κBα表达的变化。结果:宫内急性缺血缺氧20 m in时,新生大鼠Ⅱ型肺泡上皮细胞和毛细血管内皮细胞受损;肺组织TNFα-蛋白和mRNA表达明显增强(P均<0.01),κI-Bα表达明显减弱(P<0.05),并随缺血缺氧时间延长病变加重。结论:围产期急性缺血缺氧致新生大鼠肺损伤,Iκ-Bα和TNFα-的异常表达可能是肺损伤的重要原因。
