[ Objective] To investigate the mechanisms involved in the Up-regulatory effects of 17β-estrodiol on β-defensin-2 （SBD-2） in epithelial cells of ovine oviduct. [ Methods] Epithelial cells of ovine oviduct were iso...[ Objective] To investigate the mechanisms involved in the Up-regulatory effects of 17β-estrodiol on β-defensin-2 （SBD-2） in epithelial cells of ovine oviduct. [ Methods] Epithelial cells of ovine oviduct were isolated and cultured; and then the cultured cells at secondary generation were divided into 17β-estradiol （E2, 10^-8 tool/L） group, estrogen nuclear receptor antagonist ICI182780 （10^-7 tool/L） group, PKA antagonist KT-5720 （1 μmol/L） group, PKC antagonist H- 7（50 μmol/L） group, nuclear factor kappa B antagonist PDTC（50μmol/L） group and the blank control group （ Control ）. Firstly, different antagonists were added into corresponding antagonist groups in order to interfere the epithelial ceils of ovine oviduct for 1 h. Then, 17β-estradiol （ 10^-8 mol/L） was added into each antagonist group and E2 group for cultivation for 6 h. Finally, real-time fluorescent quantitative RT-PCR was used to detect the changes of SBD-2 mRNA expression. [ Results] 10^-8 mol/L 17β-estrodiol had significantly Up-regulatory effects on the expression of SBD-2 mRNA （P 〈 0. 05 ）. Estrogen nuclear receptor antagonist ICI182780, NF-κB antagonist PDTC and PKC antagonist H-7 could all block the Up-regnlatory effects on SBD-2. But PKA antagonist KT-5720 showed no significant effects on the Up-regulation of SBD-2 mRNA expression induced by 17β-estrodiol. [ Conclusions] SBD-2 mRNA expression induced by 17β-estrodiol in epithelial cells of ovine oviduct was mediated by estrogen nuclear receptor ICI182780, NF-κB and PKC pathways. However, PKA pathway might not participate in the Up-regulation of SBD-2 mRNA expression.展开更多
AIM To investigate the antitumor effects and underlying mechanisms of(17 R,18 R)-2-(1-hexyloxyethyl)-2-devinyl chlorine E6 trisodium salt(YLG-1)-induced photodynamic therapy(PDT) on pancreatic cancer in vitro and in v...AIM To investigate the antitumor effects and underlying mechanisms of(17 R,18 R)-2-(1-hexyloxyethyl)-2-devinyl chlorine E6 trisodium salt(YLG-1)-induced photodynamic therapy(PDT) on pancreatic cancer in vitro and in vivo.METHODS YLG-1 is a novel photosensitizer extracted from spirulina. Its phototoxicity, cellular uptake and localization, as well as its effect on reactive oxygen species(ROS) production, apoptosis, and expression of apoptosis-associated proteins were detected in vitro. An in vivo imaging system(IVIS), the Lumina K imaging system, and mouse models of subcutaneous Panc-1-bearing tumors were exploited to evaluate the drug delivery pathway and pancreatic cancer growth in vivo.RESULTS YLG-1 was localized to the mitochondria, and the appropriate incubation time was 6 h. Under 650 nm light irradiation, YLG-1-PDT exerted a potent cytotoxic effect on pancreatic cancer cells in vitro, which could be abolished by the ROS scavenger N-acetyl-L-cysteine(NAC). The death mode caused by YLG-1-PDT was apoptosis, accompanied by upregulated Bax and cleaved Caspase-3 and decreased Bcl-2 expression. The results from the IVIS images suggested that the optimal administration route was intratumoral(IT) injection and that the best time to conduct YLG-1-PDT was 2 h post-IT injection. Consistent with the results in vitro, YLG-1-PDT showed great growth inhibition effects on pancreatic cancer cells in a mouse model.CONCLUSION YLG-1 is a potential photosensitizer for pancreatic cancer PDT via IT injection, the mechanisms of which are associated with inducing ROS and promoting apoptosis.展开更多
基金Supported by the National Natural Science Foundation of China(31060328)the Natural Science Foundation of Inner Mongolia(No.2014BS0801)+1 种基金the Doctor's Start-up Fund in Inner Mongolia Medical University(NY2011BQ003)the Youth Entrepreneurship Foundation of Inner Mongolia Medical University(NY2010QN003)
文摘[ Objective] To investigate the mechanisms involved in the Up-regulatory effects of 17β-estrodiol on β-defensin-2 （SBD-2） in epithelial cells of ovine oviduct. [ Methods] Epithelial cells of ovine oviduct were isolated and cultured; and then the cultured cells at secondary generation were divided into 17β-estradiol （E2, 10^-8 tool/L） group, estrogen nuclear receptor antagonist ICI182780 （10^-7 tool/L） group, PKA antagonist KT-5720 （1 μmol/L） group, PKC antagonist H- 7（50 μmol/L） group, nuclear factor kappa B antagonist PDTC（50μmol/L） group and the blank control group （ Control ）. Firstly, different antagonists were added into corresponding antagonist groups in order to interfere the epithelial ceils of ovine oviduct for 1 h. Then, 17β-estradiol （ 10^-8 mol/L） was added into each antagonist group and E2 group for cultivation for 6 h. Finally, real-time fluorescent quantitative RT-PCR was used to detect the changes of SBD-2 mRNA expression. [ Results] 10^-8 mol/L 17β-estrodiol had significantly Up-regulatory effects on the expression of SBD-2 mRNA （P 〈 0. 05 ）. Estrogen nuclear receptor antagonist ICI182780, NF-κB antagonist PDTC and PKC antagonist H-7 could all block the Up-regnlatory effects on SBD-2. But PKA antagonist KT-5720 showed no significant effects on the Up-regulation of SBD-2 mRNA expression induced by 17β-estrodiol. [ Conclusions] SBD-2 mRNA expression induced by 17β-estrodiol in epithelial cells of ovine oviduct was mediated by estrogen nuclear receptor ICI182780, NF-κB and PKC pathways. However, PKA pathway might not participate in the Up-regulation of SBD-2 mRNA expression.
基金Supported by National Natural Science Foundation of China,No.81472844
文摘AIM To investigate the antitumor effects and underlying mechanisms of(17 R,18 R)-2-(1-hexyloxyethyl)-2-devinyl chlorine E6 trisodium salt(YLG-1)-induced photodynamic therapy(PDT) on pancreatic cancer in vitro and in vivo.METHODS YLG-1 is a novel photosensitizer extracted from spirulina. Its phototoxicity, cellular uptake and localization, as well as its effect on reactive oxygen species(ROS) production, apoptosis, and expression of apoptosis-associated proteins were detected in vitro. An in vivo imaging system(IVIS), the Lumina K imaging system, and mouse models of subcutaneous Panc-1-bearing tumors were exploited to evaluate the drug delivery pathway and pancreatic cancer growth in vivo.RESULTS YLG-1 was localized to the mitochondria, and the appropriate incubation time was 6 h. Under 650 nm light irradiation, YLG-1-PDT exerted a potent cytotoxic effect on pancreatic cancer cells in vitro, which could be abolished by the ROS scavenger N-acetyl-L-cysteine(NAC). The death mode caused by YLG-1-PDT was apoptosis, accompanied by upregulated Bax and cleaved Caspase-3 and decreased Bcl-2 expression. The results from the IVIS images suggested that the optimal administration route was intratumoral(IT) injection and that the best time to conduct YLG-1-PDT was 2 h post-IT injection. Consistent with the results in vitro, YLG-1-PDT showed great growth inhibition effects on pancreatic cancer cells in a mouse model.CONCLUSION YLG-1 is a potential photosensitizer for pancreatic cancer PDT via IT injection, the mechanisms of which are associated with inducing ROS and promoting apoptosis.
