Objective： To investigate the effects of Panax notoginseng saponins （PNS） on the proliferation and differentiation in NIH3T3 cells. Methods： NIH3T3 cells were treated by various concentrations of PNS 0, 0.05, 0.10...Objective： To investigate the effects of Panax notoginseng saponins （PNS） on the proliferation and differentiation in NIH3T3 cells. Methods： NIH3T3 cells were treated by various concentrations of PNS 0, 0.05, 0.10, 0.20, and 0.40 g/L. The vitality and proliferation potential of cells were detected by 3-（4,5-dimethylthiazol- 2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay, the alkaline phosphatase （ALP） activity was measured by p-nitrophenyl phosphate （pNPP） assay, and the mineralization formation ability was tested for the cellular differentiation toward osteoblast, as well as the expression level of phosphorylated extrecellular signal-regulated kinasel/2（P-ERK1/2）, extracellular signal-regulated kinasel/2 （ERK1/2） protein kinase was analyzed by Western blot with total cell lysate of NIH3T3 cells treated by PNS. Results： Both MTT and pNPP assay showed that optical density （OD） values were increased in response to PNS treatment at a dose-dependent pattern. The mineralization formation ability was enhanced in PNS-treated NIH3T3 cells compared with untreated cells. Meanwhile, the expression level of P-ERK1/2 protein kinase was up-regulated in PNS-treated NIH3T3 cells, while, the expression level of ERK1/2 protein kinase revealed no obvious difference with or without PNS treated cells. Conclusion： PNS could pay a role to promote the proliferation and differentiation in NIH3T3 cells by means of up-regulation of P-ERK1/2 protein kinase.展开更多
目的:探讨水凝胶递送碱性成纤维细胞生长因子(bFGF)对氧糖剥夺(OGD)后小鼠成纤维NIH3T3细胞功能的影响,并阐明负载bFGF的水凝胶对OGD条件下成纤维细胞氧化应激反应的改善作用。方法:制备bFGF明胶水凝胶,将对数生长期NIH3T3细胞分为空白...目的:探讨水凝胶递送碱性成纤维细胞生长因子(bFGF)对氧糖剥夺(OGD)后小鼠成纤维NIH3T3细胞功能的影响,并阐明负载bFGF的水凝胶对OGD条件下成纤维细胞氧化应激反应的改善作用。方法:制备bFGF明胶水凝胶,将对数生长期NIH3T3细胞分为空白组、20%明胶组和20%明胶+戊二醛组,采用CCK-8法检测6、12和24 h水凝胶浸出液共培养的NIH3T3细胞活性。将NIH3T3细胞分为对照组、OGD组和OGD+不同质量bFGF负载组(OGD+1 ng bFGF组、OGD+10 ng bFGF组和OGD+100 ng bFGF组)。采用Western blotting法检测各组细胞中α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白和Ⅲ型胶原蛋白表达水平。检测各组细胞中丙二醛(MDA)水平和乳酸脱氢酶(LDH)及超氧化物歧化酶(SOD)活性。采用酶联免疫吸附试验(ELISA)法检测bFGF体外释放率。结果:CCK-8法检测,与空白组比较,不同浸出时间20%明胶组和20%明胶+戊二醛组NIH3T3细胞活性差异无统计学意义(P>0.05)。Western blotting法检测,与对照组比较,OGD组NIH3T3细胞中α-SMA和Ⅰ型胶原蛋白表达水平明显升高(P<0.05或P<0.01);与OGD组比较,OGD+1 ng bFGF组、OGD+10 ng bFGF组和OGD+100 ng bFGF组NIH3T3细胞中α-SMA蛋白表达水平均明显降低(P<0.01),Ⅰ型胶原蛋白表达水平均明显降低(P<0.05或P<0.01),并呈剂量依赖性。与对照组比较,OGD组NIH3T3细胞中MDA水平和LDH活性升高(P<0.05),SOD活性降低(P<0.05)。与OGD组比较,OGD+1 ng bFGF组NIH3T3细胞中MDA水平和LDH活性降低(P<0.05),SOD活性升高(P<0.05)。ELISA法检测,在4 h内约10%bFGF由水凝胶中释放,在12 h内约15%bFGF由水凝胶中释放,在24 h内约21%bFGF由水凝胶中释放。结论:负载bFGF的水凝胶可以调控OGD条件下成纤维细胞的转化,抑制成纤维细胞Ⅰ型胶原蛋白表达,并改善细胞氧化应激反应,且该系统具有一定的持续释药能力。展开更多
基金Supported by the Grant from Zhejiang Provincial Natural Science Foundation of China(No.Y207728)Traditional Chinese Medicine Adiministration Bureau of Zhejiang Province, China(No.2008YA005)
文摘Objective： To investigate the effects of Panax notoginseng saponins （PNS） on the proliferation and differentiation in NIH3T3 cells. Methods： NIH3T3 cells were treated by various concentrations of PNS 0, 0.05, 0.10, 0.20, and 0.40 g/L. The vitality and proliferation potential of cells were detected by 3-（4,5-dimethylthiazol- 2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay, the alkaline phosphatase （ALP） activity was measured by p-nitrophenyl phosphate （pNPP） assay, and the mineralization formation ability was tested for the cellular differentiation toward osteoblast, as well as the expression level of phosphorylated extrecellular signal-regulated kinasel/2（P-ERK1/2）, extracellular signal-regulated kinasel/2 （ERK1/2） protein kinase was analyzed by Western blot with total cell lysate of NIH3T3 cells treated by PNS. Results： Both MTT and pNPP assay showed that optical density （OD） values were increased in response to PNS treatment at a dose-dependent pattern. The mineralization formation ability was enhanced in PNS-treated NIH3T3 cells compared with untreated cells. Meanwhile, the expression level of P-ERK1/2 protein kinase was up-regulated in PNS-treated NIH3T3 cells, while, the expression level of ERK1/2 protein kinase revealed no obvious difference with or without PNS treated cells. Conclusion： PNS could pay a role to promote the proliferation and differentiation in NIH3T3 cells by means of up-regulation of P-ERK1/2 protein kinase.
文摘目的:探讨水凝胶递送碱性成纤维细胞生长因子(bFGF)对氧糖剥夺(OGD)后小鼠成纤维NIH3T3细胞功能的影响,并阐明负载bFGF的水凝胶对OGD条件下成纤维细胞氧化应激反应的改善作用。方法:制备bFGF明胶水凝胶,将对数生长期NIH3T3细胞分为空白组、20%明胶组和20%明胶+戊二醛组,采用CCK-8法检测6、12和24 h水凝胶浸出液共培养的NIH3T3细胞活性。将NIH3T3细胞分为对照组、OGD组和OGD+不同质量bFGF负载组(OGD+1 ng bFGF组、OGD+10 ng bFGF组和OGD+100 ng bFGF组)。采用Western blotting法检测各组细胞中α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白和Ⅲ型胶原蛋白表达水平。检测各组细胞中丙二醛(MDA)水平和乳酸脱氢酶(LDH)及超氧化物歧化酶(SOD)活性。采用酶联免疫吸附试验(ELISA)法检测bFGF体外释放率。结果:CCK-8法检测,与空白组比较,不同浸出时间20%明胶组和20%明胶+戊二醛组NIH3T3细胞活性差异无统计学意义(P>0.05)。Western blotting法检测,与对照组比较,OGD组NIH3T3细胞中α-SMA和Ⅰ型胶原蛋白表达水平明显升高(P<0.05或P<0.01);与OGD组比较,OGD+1 ng bFGF组、OGD+10 ng bFGF组和OGD+100 ng bFGF组NIH3T3细胞中α-SMA蛋白表达水平均明显降低(P<0.01),Ⅰ型胶原蛋白表达水平均明显降低(P<0.05或P<0.01),并呈剂量依赖性。与对照组比较,OGD组NIH3T3细胞中MDA水平和LDH活性升高(P<0.05),SOD活性降低(P<0.05)。与OGD组比较,OGD+1 ng bFGF组NIH3T3细胞中MDA水平和LDH活性降低(P<0.05),SOD活性升高(P<0.05)。ELISA法检测,在4 h内约10%bFGF由水凝胶中释放,在12 h内约15%bFGF由水凝胶中释放,在24 h内约21%bFGF由水凝胶中释放。结论:负载bFGF的水凝胶可以调控OGD条件下成纤维细胞的转化,抑制成纤维细胞Ⅰ型胶原蛋白表达,并改善细胞氧化应激反应,且该系统具有一定的持续释药能力。
