目的:观察雌激素(E2)诱导的大鼠慢性非细菌性前列腺炎(chronic nonbacterial prostatitis,CNP)中信号转导子及转录激活子1(signal transducer and activator of transcription 1,STAT-1)的激活和调节活化正常T细胞表达及分泌因子(regula...目的:观察雌激素(E2)诱导的大鼠慢性非细菌性前列腺炎(chronic nonbacterial prostatitis,CNP)中信号转导子及转录激活子1(signal transducer and activator of transcription 1,STAT-1)的激活和调节活化正常T细胞表达及分泌因子(regulated on activation,normal T-cell expressed and secreted,RANTES)的表达,探讨雌激素诱导炎症形成的机制。方法:将80只老年雄性SD大鼠随机分为对照组、去势组、去势+E2组和去势+E2+AG490组,每组20只。苏木素-伊红(HE)染色观察大鼠前列腺组织病理改变。Western blotting法测定STAT-1及p-STAT-1蛋白水平。RT-PCR和免疫组化SP法分别检测RANTES mRNA和蛋白表达水平,并分析p-STAT-1与RANTES表达水平的相关性。结果:去势+E2组前列腺组织呈明显炎症表现。去势+E2组中STAT-1及p-STAT-1蛋白表达明显高于对照组及去势组(P<0.01),RANTES mRNA和蛋白表达也显著增高(P<0.01)。去势+E2+AG490组中STAT-1及p-STAT-1蛋白、RANTES mRNA和蛋白表达水平较去势+E2组明显降低(P<0.01)。大鼠前列腺组织中p-STAT-1表达水平与RANTES mRNA转录水平呈正相关(r=0.735,P<0.05),RANTES表达水平与RANTESmRNA转录水平亦呈正相关(r=0.694,P<0.05)。结论:雌激素诱导CNP的形成可能与调节转录因子STAT-1、促进RANTES分子的表达及炎症反应有关。展开更多
In order to improve the extracellular endo-1,4-β-mannosidase(MAN) activity of recombinant Pichia pastoris, optimization of signal peptides was investigated. At first, five potential signal peptides(W1, MF4 I, INU1 A,...In order to improve the extracellular endo-1,4-β-mannosidase(MAN) activity of recombinant Pichia pastoris, optimization of signal peptides was investigated. At first, five potential signal peptides(W1, MF4 I, INU1 A, αpre, HFBI) were chosen to be analyzed by Signal P 4.0, among which W1 was designed. Then, the widely used signal peptide α-factor in expression vector p GAPZαA was replaced by those five signal peptides to reconstruct five new expression vectors. MAN activity was assayed after expression vectors were transformed into Pichia pastoris. The data show that the relative efficiencies of W1, MF4 I, INU1 A, αpre, and HFBI signal peptides are 23.5%, 203.5%, 0, 79.7%, and 120.3% compared with α-factor, respectively. The further gene copy number determination by the quantitative real-time PCR reveals that the MAN activities mediated by α-factor from 1 to 6 gene copy number levels are 12.95, 43.33, 126.63, 173.53, 103.23 and 88.63 U/m L, while those mediated by MF4 I are 79.22, 133.89, 260.14, 347.5, 206.15 and 181.89 U/m L, respectively. The maximum MAN activity reached 347.5 U/m L with 4 gene copies mediated by MF4 I. These results indicate that replacing the signal peptide α-factor with MF4 I and increasing MAN gene copies to a proper number can greatly improve the secretory expression of MAN.展开更多
文摘目的:观察雌激素(E2)诱导的大鼠慢性非细菌性前列腺炎(chronic nonbacterial prostatitis,CNP)中信号转导子及转录激活子1(signal transducer and activator of transcription 1,STAT-1)的激活和调节活化正常T细胞表达及分泌因子(regulated on activation,normal T-cell expressed and secreted,RANTES)的表达,探讨雌激素诱导炎症形成的机制。方法:将80只老年雄性SD大鼠随机分为对照组、去势组、去势+E2组和去势+E2+AG490组,每组20只。苏木素-伊红(HE)染色观察大鼠前列腺组织病理改变。Western blotting法测定STAT-1及p-STAT-1蛋白水平。RT-PCR和免疫组化SP法分别检测RANTES mRNA和蛋白表达水平,并分析p-STAT-1与RANTES表达水平的相关性。结果:去势+E2组前列腺组织呈明显炎症表现。去势+E2组中STAT-1及p-STAT-1蛋白表达明显高于对照组及去势组(P<0.01),RANTES mRNA和蛋白表达也显著增高(P<0.01)。去势+E2+AG490组中STAT-1及p-STAT-1蛋白、RANTES mRNA和蛋白表达水平较去势+E2组明显降低(P<0.01)。大鼠前列腺组织中p-STAT-1表达水平与RANTES mRNA转录水平呈正相关(r=0.735,P<0.05),RANTES表达水平与RANTESmRNA转录水平亦呈正相关(r=0.694,P<0.05)。结论:雌激素诱导CNP的形成可能与调节转录因子STAT-1、促进RANTES分子的表达及炎症反应有关。
基金Project(13JJ9002)supported by Hunan Provincial Natural Science Foundation of ChinaProject(2012XK4081)supported by the Key Science Technology Plan Project of Hunan Provincial Science&Technology Department,ChinaProject(CX2012B124)supported by the Graduate Degree Thesis Innovation Program of Hunan Province,China
文摘In order to improve the extracellular endo-1,4-β-mannosidase(MAN) activity of recombinant Pichia pastoris, optimization of signal peptides was investigated. At first, five potential signal peptides(W1, MF4 I, INU1 A, αpre, HFBI) were chosen to be analyzed by Signal P 4.0, among which W1 was designed. Then, the widely used signal peptide α-factor in expression vector p GAPZαA was replaced by those five signal peptides to reconstruct five new expression vectors. MAN activity was assayed after expression vectors were transformed into Pichia pastoris. The data show that the relative efficiencies of W1, MF4 I, INU1 A, αpre, and HFBI signal peptides are 23.5%, 203.5%, 0, 79.7%, and 120.3% compared with α-factor, respectively. The further gene copy number determination by the quantitative real-time PCR reveals that the MAN activities mediated by α-factor from 1 to 6 gene copy number levels are 12.95, 43.33, 126.63, 173.53, 103.23 and 88.63 U/m L, while those mediated by MF4 I are 79.22, 133.89, 260.14, 347.5, 206.15 and 181.89 U/m L, respectively. The maximum MAN activity reached 347.5 U/m L with 4 gene copies mediated by MF4 I. These results indicate that replacing the signal peptide α-factor with MF4 I and increasing MAN gene copies to a proper number can greatly improve the secretory expression of MAN.
