A selective, sensitive, and convenient assay for human collagenase is required because of its implication in diseases such as rheumatoid arthritis, osteoarthritis, and tumors. Here, a novel assay for human collagenase...A selective, sensitive, and convenient assay for human collagenase is required because of its implication in diseases such as rheumatoid arthritis, osteoarthritis, and tumors. Here, a novel assay for human collagenase activity is described in which enzymatic degradation of native collagen or acetyl peptide is determined by using a fluorogenic reaction for oligopeptides. The oligopeptides are quantified spectrofluorometrically with either 3,4-dihydroxyphenylacetic acid or 1,2-dihydroxybenzen reaction in the presence of sodium periodate and sodium borate (pH 7 - 8). These reactions can selectively convert N-terminal Gly-containing oligopeptides and N-terminal Ile-containing oligopeptides to fluorescence (FL) compounds, respectively, but not proteins, acetyl peptides or amino acids. Under optimized conditions using 1.65 μM collagen IV or 1.5 mM Ac-GPQGI- AGQ as substrates, this assay exhibits a proportional relationship between FL intensities and human collagenase-3 (MMP-13) concentrations. It can assay endogenous collagenase activities in several biological samples, such as cultured human cells and cheek tissue.展开更多
背景:腰椎间盘退行性疾病是导致腰痛和下肢神经症状的常见疾病。胶原代谢的平衡在维持椎间盘的稳定中发挥着重要的作用。目的:对椎间盘退变中胶原蛋白代谢失衡的研究进展作一综述。方法:由第一作者在中国知网、PubMed、Web of Science...背景:腰椎间盘退行性疾病是导致腰痛和下肢神经症状的常见疾病。胶原代谢的平衡在维持椎间盘的稳定中发挥着重要的作用。目的:对椎间盘退变中胶原蛋白代谢失衡的研究进展作一综述。方法:由第一作者在中国知网、PubMed、Web of Science数据库中检索2024年5月之前发表的相关文章,中文检索词为“椎间盘退变疾病”“胶原代谢”“胶原酶家族”“胶原合成相关因子”“细胞因子”,英文检索词为“degenerative disc disease”“Collagen metabolism”“Collagenase family”“Collagen synthesis related factors”“Collagen breakdown related factors”。最终纳入76篇文献进行综述。结果与结论:在椎间盘退变的过程中,胶原蛋白代谢的平衡在维持正常椎间盘的稳定中起到至关重要的作用。当椎间盘发生退变时,椎间盘内分泌大量促炎因子、胶原酶,并发生氧化应激反应等,使椎间盘内胶原蛋白分解增加,同时抑制生长因子、胶原蛋白合成酶、胶原蛋合成相关因子的生成,使椎间盘内胶原蛋白合成减少。上述两种情况共同作用使椎间盘内胶原蛋白的代谢平衡被打破,进一步加剧了椎间盘退变的进程。展开更多
INTRODUCTIONLiver fibrosis is a dynamic course leading tocirrhosis from a various chronic liver diseases. Thepathological basis of fibrosis is the disturbance ofproduction and degradation of the extracellularmatrix (E...INTRODUCTIONLiver fibrosis is a dynamic course leading tocirrhosis from a various chronic liver diseases. Thepathological basis of fibrosis is the disturbance ofproduction and degradation of the extracellularmatrix (ECM), which causes accumulation of ECMin the liver[1,2].展开更多
Insulin-like growth factor 1 （IGF-I） is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half- life, rapid clearance, and limited target specificity. To achieve t...Insulin-like growth factor 1 （IGF-I） is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half- life, rapid clearance, and limited target specificity. To achieve targeted and long-lasting treatment, we investigated the addition of a binding structure by fusing a collagen-binding domain to IGF- 1. After confirming its affinity for collagen, the biological activity of this construct was examined by measuring cell proliferation after transfection into PC12 and Schwann cells using a 3-（4,5-dimethyl-2-thiazolyl）-2,5-di- phenyl-2-H-tetrazolium bromide assay. Immunofluorescence staining was conducted to detect neurofilament and microtubule-associated protein 2 expression, while real time-polymerase chain reaction was utilized to determine IGF-1 receptor and nerve growth/actor mRNA expression. Our results demonstrate a significant increase in collagen-binding activity of the recombinant protein compared with IGF-1. Moreover, the recombinant protein promoted proliferation of PC12 and Schwann cells, and increased the expression of neurofilament and microtubule-associated protein 2. Importantly, the recombinant protein also stimulated sustained expression of IGF-1 receptor and nerve growth factor mRNA for days. These results show that the recombinant protein achieved the goal of targeting and long-lasting treatment, and thus could become a clinically used factor for promoting nerve regeneration with a prolonged therapeutic effect.展开更多
文摘A selective, sensitive, and convenient assay for human collagenase is required because of its implication in diseases such as rheumatoid arthritis, osteoarthritis, and tumors. Here, a novel assay for human collagenase activity is described in which enzymatic degradation of native collagen or acetyl peptide is determined by using a fluorogenic reaction for oligopeptides. The oligopeptides are quantified spectrofluorometrically with either 3,4-dihydroxyphenylacetic acid or 1,2-dihydroxybenzen reaction in the presence of sodium periodate and sodium borate (pH 7 - 8). These reactions can selectively convert N-terminal Gly-containing oligopeptides and N-terminal Ile-containing oligopeptides to fluorescence (FL) compounds, respectively, but not proteins, acetyl peptides or amino acids. Under optimized conditions using 1.65 μM collagen IV or 1.5 mM Ac-GPQGI- AGQ as substrates, this assay exhibits a proportional relationship between FL intensities and human collagenase-3 (MMP-13) concentrations. It can assay endogenous collagenase activities in several biological samples, such as cultured human cells and cheek tissue.
文摘背景:腰椎间盘退行性疾病是导致腰痛和下肢神经症状的常见疾病。胶原代谢的平衡在维持椎间盘的稳定中发挥着重要的作用。目的:对椎间盘退变中胶原蛋白代谢失衡的研究进展作一综述。方法:由第一作者在中国知网、PubMed、Web of Science数据库中检索2024年5月之前发表的相关文章,中文检索词为“椎间盘退变疾病”“胶原代谢”“胶原酶家族”“胶原合成相关因子”“细胞因子”,英文检索词为“degenerative disc disease”“Collagen metabolism”“Collagenase family”“Collagen synthesis related factors”“Collagen breakdown related factors”。最终纳入76篇文献进行综述。结果与结论:在椎间盘退变的过程中,胶原蛋白代谢的平衡在维持正常椎间盘的稳定中起到至关重要的作用。当椎间盘发生退变时,椎间盘内分泌大量促炎因子、胶原酶,并发生氧化应激反应等,使椎间盘内胶原蛋白分解增加,同时抑制生长因子、胶原蛋白合成酶、胶原蛋合成相关因子的生成,使椎间盘内胶原蛋白合成减少。上述两种情况共同作用使椎间盘内胶原蛋白的代谢平衡被打破,进一步加剧了椎间盘退变的进程。
基金Project supported by the National Natural Science Foundation of China, No. 39500138
文摘INTRODUCTIONLiver fibrosis is a dynamic course leading tocirrhosis from a various chronic liver diseases. Thepathological basis of fibrosis is the disturbance ofproduction and degradation of the extracellularmatrix (ECM), which causes accumulation of ECMin the liver[1,2].
基金supported by the National Natural Science Foundation of China,No.81350013a grant from the Jilin Provincial Science and Technology Plan of China,No.20160101027JC&SC201502001the Graduate Innovation Fund of Jilin University in China,No.2017031&2017176
文摘Insulin-like growth factor 1 （IGF-I） is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half- life, rapid clearance, and limited target specificity. To achieve targeted and long-lasting treatment, we investigated the addition of a binding structure by fusing a collagen-binding domain to IGF- 1. After confirming its affinity for collagen, the biological activity of this construct was examined by measuring cell proliferation after transfection into PC12 and Schwann cells using a 3-（4,5-dimethyl-2-thiazolyl）-2,5-di- phenyl-2-H-tetrazolium bromide assay. Immunofluorescence staining was conducted to detect neurofilament and microtubule-associated protein 2 expression, while real time-polymerase chain reaction was utilized to determine IGF-1 receptor and nerve growth/actor mRNA expression. Our results demonstrate a significant increase in collagen-binding activity of the recombinant protein compared with IGF-1. Moreover, the recombinant protein promoted proliferation of PC12 and Schwann cells, and increased the expression of neurofilament and microtubule-associated protein 2. Importantly, the recombinant protein also stimulated sustained expression of IGF-1 receptor and nerve growth factor mRNA for days. These results show that the recombinant protein achieved the goal of targeting and long-lasting treatment, and thus could become a clinically used factor for promoting nerve regeneration with a prolonged therapeutic effect.
