目的:观察表皮生长因子(EGF)对胰腺癌细胞KP4增殖、黏附及侵袭力和核转录因子(NF-κB)、尿激酶型纤溶酶原激活物(uPA)表达的影响．方法:通过细胞侵袭、增殖及黏附实验观察在EGF影响下肿瘤细胞侵袭、增殖及黏附能力变化．Western blot,RT...目的:观察表皮生长因子(EGF)对胰腺癌细胞KP4增殖、黏附及侵袭力和核转录因子(NF-κB)、尿激酶型纤溶酶原激活物(uPA)表达的影响．方法:通过细胞侵袭、增殖及黏附实验观察在EGF影响下肿瘤细胞侵袭、增殖及黏附能力变化．Western blot,RT-PCR及EMSA实验检测胰腺癌细胞的NF-κB活性和uPA表达并观察在NF-κB抑制物PDTC抑制下EGF诱导的NF-κB活性和uPA表达及肿瘤细胞侵袭力变化．结果:EGF能够明显促进胰腺癌细胞的侵袭能力,具有明显的剂量依赖性(50,25,5μg/L vs 0μg/L:116±13,97±10,83±7 vs 72±5;t= 3．552,3．018,2．373;P=0．006,0．015,0．042),然而对增殖及黏附力无明显影响．随着EGF浓度的增加,NF-κB活性明显增强．EGF上调uPA蛋白及mRNA表达,具有浓度依赖性．PDTC显著抑制EGF诱导的NF-κB活性、uPA表达及胰腺癌细胞的侵袭力．结论:EGF通过激活NF-κB而诱导uPA表达,促进胰腺癌细胞的侵袭和转移．展开更多
Objective：To investigate the mechanism of action of Fuzheng Huayu Formula（扶正化瘀方,FZHY）against renal interstitial fibrosis（RIF）relating to oxidative injury and nuclear factor-kappa B（NF-κB）activity.Methods...Objective：To investigate the mechanism of action of Fuzheng Huayu Formula（扶正化瘀方,FZHY）against renal interstitial fibrosis（RIF）relating to oxidative injury and nuclear factor-kappa B（NF-κB）activity.Methods：Thirty-two Sprague-Dawley rats were randomly divided into 3 groups：normal group,model group and FZHY treatment group.The RIF model was induced by oral administration of HgC l2 at a dose of 8 mg/kg body weight once a day for 9 weeks.Meanwhile,rats in FZHY treatment group orally took FZHY at a dose of4.0 g/kg rat weight for 9 weeks.The content of hydroxyproline（Hyp）and collagen deposition in kidney were observed.The activities of superoxide dismutase（SOD）and glutathione peroxidase（GSH-Px）,the content of glutathione（GSH）and malondialdehyde（MDA）of kidney were tested.The expressions of inhibitor-κappa B（IκB）,phospho-IκB（p-IκB）,tumor necrosis factor-α（TNF-α）,matrix metalloproteinase-2（MMP-2）andα-smooth muscle actin（α-SMA）were analyzed by Western blot.α-SMA expression was also observed by immunofluorescent staining.MMP-2 activity was measured by gelatin zymography.NF-κB activation was determined by electrophoretic mobility shift assay.Results：Renal interstitial fibrosis was induced by Hg Cl2,demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney（P〈0.01）.FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the Hg Cl2-treated rats（P〈0.01）.GSH content decreased obviously,and MDA content increased significantly in HgC l2-treated rats compared with that of normal rats（P〈0.01）.FZHY significantly increased GSH content and decreased MDA content in the model rats（P〈0.01）.The expressionα-SMA was increased in model rats compared with that of normal rats,FZHY significantly decreased its expression（P〈0.01）.The expressions of p-IκB and TNF-αand MMP-2,MMP-2 activity,and NF-κB activation were increased in model group compared with that in normal group（P〈0.01）,FZHY significantly decreased NF-κB activation,MMP-2 activity and p-IκB and TNF-αexpressions（P〈0.01）.Conclusions：FZHY could protect kidney from oxidative injury intoxicated by Hg Cl2,and antagonized oxidative stress-stimulated NF-κB activity through inhibition of IκB phosphorylation in the interstitial fibrotic kidney,these effects importantly contributed to FZHY action mechanism against renal interstitial fibrosis.展开更多
文摘目的:观察表皮生长因子(EGF)对胰腺癌细胞KP4增殖、黏附及侵袭力和核转录因子(NF-κB)、尿激酶型纤溶酶原激活物(uPA)表达的影响．方法:通过细胞侵袭、增殖及黏附实验观察在EGF影响下肿瘤细胞侵袭、增殖及黏附能力变化．Western blot,RT-PCR及EMSA实验检测胰腺癌细胞的NF-κB活性和uPA表达并观察在NF-κB抑制物PDTC抑制下EGF诱导的NF-κB活性和uPA表达及肿瘤细胞侵袭力变化．结果:EGF能够明显促进胰腺癌细胞的侵袭能力,具有明显的剂量依赖性(50,25,5μg/L vs 0μg/L:116±13,97±10,83±7 vs 72±5;t= 3．552,3．018,2．373;P=0．006,0．015,0．042),然而对增殖及黏附力无明显影响．随着EGF浓度的增加,NF-κB活性明显增强．EGF上调uPA蛋白及mRNA表达,具有浓度依赖性．PDTC显著抑制EGF诱导的NF-κB活性、uPA表达及胰腺癌细胞的侵袭力．结论:EGF通过激活NF-κB而诱导uPA表达,促进胰腺癌细胞的侵袭和转移．
基金Supported by the National Natural Science Foundation of China(No.81270053)the National Science and Technology Major Project(No.2014ZX10005001)+1 种基金the International S&T Cooperation Program of China(No.2014DFA31440)"Three-Year Action Plan" for Development of TCM in Shanghai(No.ZY3-CCCX-2-1003)
文摘Objective：To investigate the mechanism of action of Fuzheng Huayu Formula（扶正化瘀方,FZHY）against renal interstitial fibrosis（RIF）relating to oxidative injury and nuclear factor-kappa B（NF-κB）activity.Methods：Thirty-two Sprague-Dawley rats were randomly divided into 3 groups：normal group,model group and FZHY treatment group.The RIF model was induced by oral administration of HgC l2 at a dose of 8 mg/kg body weight once a day for 9 weeks.Meanwhile,rats in FZHY treatment group orally took FZHY at a dose of4.0 g/kg rat weight for 9 weeks.The content of hydroxyproline（Hyp）and collagen deposition in kidney were observed.The activities of superoxide dismutase（SOD）and glutathione peroxidase（GSH-Px）,the content of glutathione（GSH）and malondialdehyde（MDA）of kidney were tested.The expressions of inhibitor-κappa B（IκB）,phospho-IκB（p-IκB）,tumor necrosis factor-α（TNF-α）,matrix metalloproteinase-2（MMP-2）andα-smooth muscle actin（α-SMA）were analyzed by Western blot.α-SMA expression was also observed by immunofluorescent staining.MMP-2 activity was measured by gelatin zymography.NF-κB activation was determined by electrophoretic mobility shift assay.Results：Renal interstitial fibrosis was induced by Hg Cl2,demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney（P〈0.01）.FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the Hg Cl2-treated rats（P〈0.01）.GSH content decreased obviously,and MDA content increased significantly in HgC l2-treated rats compared with that of normal rats（P〈0.01）.FZHY significantly increased GSH content and decreased MDA content in the model rats（P〈0.01）.The expressionα-SMA was increased in model rats compared with that of normal rats,FZHY significantly decreased its expression（P〈0.01）.The expressions of p-IκB and TNF-αand MMP-2,MMP-2 activity,and NF-κB activation were increased in model group compared with that in normal group（P〈0.01）,FZHY significantly decreased NF-κB activation,MMP-2 activity and p-IκB and TNF-αexpressions（P〈0.01）.Conclusions：FZHY could protect kidney from oxidative injury intoxicated by Hg Cl2,and antagonized oxidative stress-stimulated NF-κB activity through inhibition of IκB phosphorylation in the interstitial fibrotic kidney,these effects importantly contributed to FZHY action mechanism against renal interstitial fibrosis.
