Erectile dysfunction （ED） is a major complication of diabetes mellitus. Icariin has been shown to enhance erectile function through its bioactive form, icarisid Ih This study investigates the effects of icarisid Ⅱ ...Erectile dysfunction （ED） is a major complication of diabetes mellitus. Icariin has been shown to enhance erectile function through its bioactive form, icarisid Ih This study investigates the effects of icarisid Ⅱ on diabetic rats with ED and its potential mechanism viathe assessment of advanced glycosylation end products （AGEs）, autophagy, mTOR and the NO-cGMP pathway. Icarisid Ⅱ was extracted from icariin by an enzymatic method. In the control and diabetic ED groups, rats were administered normal saline; in the icarisid Ⅱ group, rats were administered icarisid Ⅱ intragastrically. Erectile function was evaluated by measuring intracavernosal pressure/mean arterial pressure （ICP/MAP）. AGE concentrations, nitric oxide synthase （NOS） activity and cGMP concentration were assessed by enzyme immunoassay. Cell proliferation was analysed using methyl thiazolyl tetrazolium assay and flow cytometry. Autophagosomes were observed by transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 Iocalisation. The expression of NOS isoforms and key proteins in autophagy were examined by western blot. Our results have shown that Icarisid Ⅱ increased ICP/MAP values, the smooth muscle cell （SMC） growth curve, S phase and SMC/collagen fibril （SMC/CF） proportions and decreased Beclin 1 （P〈0.05）. Icarisid Ⅱ significantly increased the proliferative index and p-p70S6K（Thr389） levels and decreased the numbers of autophagosomes and the levels of LC3-11 （P〈0.01）. Icarisid Ⅱ decreased AGE concentrations and increased cGMP concentration, NOS activity （P〈0.05） and cNOS levels （P〈0.01） in the diabetic ED group. Therefore, Icarisid Ⅱ constitutes a promising compound for diabetic ED and might be involved in the upregulation of SMC proliferation and the NO-cGMP pathway and the downregulation of AGEs, autophagy and the mTOR pathway.展开更多
目的：观察白子菜对胰岛素抵抗人肝癌HepG2细胞的胰岛素受体（insulin receptor，InsR），葡萄糖转运蛋白4（glucosetransporter4，GLUT4）mRNA表达和蛋白激酶B（proteinkinaseB，PKB），糖原合成激酶-3／3（glycogensynthasekinase-3β...目的：观察白子菜对胰岛素抵抗人肝癌HepG2细胞的胰岛素受体（insulin receptor，InsR），葡萄糖转运蛋白4（glucosetransporter4，GLUT4）mRNA表达和蛋白激酶B（proteinkinaseB，PKB），糖原合成激酶-3／3（glycogensynthasekinase-3β，GSK一3／3）蛋白表达的影响，探讨其干预胰岛素抵抗的分子机制。方法：用胰岛素诱导HepG2细胞使其产生胰岛素抵抗，实验分为空白组，模型组，自子菜水提物（Gynura divaricata hot water extracts，GDE）高、低质量浓度（1．0，0．5g·L^-1）组，白子菜总黄酮（G．divaricataflavonoids，GDF）高、低质量浓度（0．2，0．1g·L^-1）组，白子菜总生物碱（G．divaricata alkaloid，GDA）高、低质量浓度（0．1，0．05g·L^-1）组，二甲双胍（1×10^-3mol·L^-1）组，葡萄糖氧化酶法检测HepG2细胞培养液上清液中葡萄糖的含量，实时荧光定量聚合酶反应（Real-timePCR）检测HepG2细胞InsR，GLUT4mRNA表达，蛋白免疫印迹法（Western blot）检测HepG2细胞PKB，GSK-3β的蛋白表达。结果：与模型组比较，1．0，0．5g·L。GDE组上清液葡萄糖含量极显著降低（P〈0．01），0．2g·L^-1GDF组上清液葡萄糖含量显著降低（P〈0．05）；GDE，GDF组InsR，GLUT4的mRNA表达显著增加（P〈0．05）；GDE组PKB的蛋白表达显著增加（P〈0．05），GSK-3β的蛋白表达显著降低（P〈0．05），GDF组PKB的蛋白表达极显著增加（P〈0．01），GSK-3β的蛋白表达极显著降低（P〈0．01）。结论：白子菜可改善胰岛素抵抗HepG2细胞对葡萄糖的摄取，其机制可能与上调胰岛素抵抗HepG2细胞InsR，GLUT4mRNA表达和PKB的蛋白表达及降低GSK-3β的蛋白表达相关。展开更多
文摘Erectile dysfunction （ED） is a major complication of diabetes mellitus. Icariin has been shown to enhance erectile function through its bioactive form, icarisid Ih This study investigates the effects of icarisid Ⅱ on diabetic rats with ED and its potential mechanism viathe assessment of advanced glycosylation end products （AGEs）, autophagy, mTOR and the NO-cGMP pathway. Icarisid Ⅱ was extracted from icariin by an enzymatic method. In the control and diabetic ED groups, rats were administered normal saline; in the icarisid Ⅱ group, rats were administered icarisid Ⅱ intragastrically. Erectile function was evaluated by measuring intracavernosal pressure/mean arterial pressure （ICP/MAP）. AGE concentrations, nitric oxide synthase （NOS） activity and cGMP concentration were assessed by enzyme immunoassay. Cell proliferation was analysed using methyl thiazolyl tetrazolium assay and flow cytometry. Autophagosomes were observed by transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 Iocalisation. The expression of NOS isoforms and key proteins in autophagy were examined by western blot. Our results have shown that Icarisid Ⅱ increased ICP/MAP values, the smooth muscle cell （SMC） growth curve, S phase and SMC/collagen fibril （SMC/CF） proportions and decreased Beclin 1 （P〈0.05）. Icarisid Ⅱ significantly increased the proliferative index and p-p70S6K（Thr389） levels and decreased the numbers of autophagosomes and the levels of LC3-11 （P〈0.01）. Icarisid Ⅱ decreased AGE concentrations and increased cGMP concentration, NOS activity （P〈0.05） and cNOS levels （P〈0.01） in the diabetic ED group. Therefore, Icarisid Ⅱ constitutes a promising compound for diabetic ED and might be involved in the upregulation of SMC proliferation and the NO-cGMP pathway and the downregulation of AGEs, autophagy and the mTOR pathway.
文摘目的：观察白子菜对胰岛素抵抗人肝癌HepG2细胞的胰岛素受体（insulin receptor，InsR），葡萄糖转运蛋白4（glucosetransporter4，GLUT4）mRNA表达和蛋白激酶B（proteinkinaseB，PKB），糖原合成激酶-3／3（glycogensynthasekinase-3β，GSK一3／3）蛋白表达的影响，探讨其干预胰岛素抵抗的分子机制。方法：用胰岛素诱导HepG2细胞使其产生胰岛素抵抗，实验分为空白组，模型组，自子菜水提物（Gynura divaricata hot water extracts，GDE）高、低质量浓度（1．0，0．5g·L^-1）组，白子菜总黄酮（G．divaricataflavonoids，GDF）高、低质量浓度（0．2，0．1g·L^-1）组，白子菜总生物碱（G．divaricata alkaloid，GDA）高、低质量浓度（0．1，0．05g·L^-1）组，二甲双胍（1×10^-3mol·L^-1）组，葡萄糖氧化酶法检测HepG2细胞培养液上清液中葡萄糖的含量，实时荧光定量聚合酶反应（Real-timePCR）检测HepG2细胞InsR，GLUT4mRNA表达，蛋白免疫印迹法（Western blot）检测HepG2细胞PKB，GSK-3β的蛋白表达。结果：与模型组比较，1．0，0．5g·L。GDE组上清液葡萄糖含量极显著降低（P〈0．01），0．2g·L^-1GDF组上清液葡萄糖含量显著降低（P〈0．05）；GDE，GDF组InsR，GLUT4的mRNA表达显著增加（P〈0．05）；GDE组PKB的蛋白表达显著增加（P〈0．05），GSK-3β的蛋白表达显著降低（P〈0．05），GDF组PKB的蛋白表达极显著增加（P〈0．01），GSK-3β的蛋白表达极显著降低（P〈0．01）。结论：白子菜可改善胰岛素抵抗HepG2细胞对葡萄糖的摄取，其机制可能与上调胰岛素抵抗HepG2细胞InsR，GLUT4mRNA表达和PKB的蛋白表达及降低GSK-3β的蛋白表达相关。
