Several metabolic gene expressions are regulated in concert with muscle glycogen status. We hypothesized that intermittent exercise performed at high but sub-maximal intensities with long recovery periods would induce...Several metabolic gene expressions are regulated in concert with muscle glycogen status. We hypothesized that intermittent exercise performed at high but sub-maximal intensities with long recovery periods would induce a low glycogen state that would stimul- ate peroxisome proliferator-activated receptor-γ coa- ctivator-1α (PGC1-α) and pyruvate dehydrogenase kinase-4 (PDK-4) gene expression in muscle. Nine young human subjects performed two intermittent exercise sessions. One session consisted of 60 s cycling bouts at VO2max (IE100%), and the other session consisted of 75 s cycling bouts at 80% VO2max (IE80%). Twelve bouts of exercise were completed in both sessions with a 4 min rest between each bout. Muscle specimens were obtained at pre-exercise and immediately, 1.5 h and 3 h post-exercise. Muscle gly- cogen was significantly decreased after both sessions (IE100%, 94.1 ± 5.8 to 38.7 ± 5.5 mmol/kg w.w.;IE80%, 94.6 ± 9.1 to 53.3 ± 4.8 mmol/kg w.w.;both P α and PDK- 4 mRNA expression were significantly increased after exercise in both IE100% and IE80% (PGC-1α: ~3.7 and ~2.9-fold, respectively;PDK-4: ~11.1 and ~3.5-fold, respectively;all P 100% than in IE80% (P a and PDK-4 mRNA expression, suggesting that increasing exercise intensity contributes to muscle glycogen depletion and PDK-4 mRNA expression in human skeletal muscle.展开更多
目的探讨脊髓背角星形胶质细胞中丙酮酸脱氢酶激酶4(PDK4)在吗啡耐受小鼠中的作用。方法SPF级雄性C57b1/6小鼠24只,6~8周龄,体重22~25 g。采用随机数字表法分为四组:生理盐水组(C组)、吗啡组(M组)、PDK4抑制剂二氯乙酸(DCA)组(D组)和吗...目的探讨脊髓背角星形胶质细胞中丙酮酸脱氢酶激酶4(PDK4)在吗啡耐受小鼠中的作用。方法SPF级雄性C57b1/6小鼠24只,6~8周龄,体重22~25 g。采用随机数字表法分为四组:生理盐水组(C组)、吗啡组(M组)、PDK4抑制剂二氯乙酸(DCA)组(D组)和吗啡+DCA组四组(MD组),每组6只。C组鞘内注射生理盐水10μl,每日2次,连续5 d;M组、D组和MD组分别鞘内注射含吗啡10μg、DCA 200μg和吗啡10μg+DCA 200μg的生理盐水10μl,每日2次,连续5 d。首次给药前和每日第1次给药后1 h检测小鼠甩尾潜伏期(TFL)。最后一次给药后处死小鼠,采用Western blot法检测脊髓背角中PDK4、磷酸化丙酮酸脱氢酶(p-PDH)、p-S^(293)-PDH、p-S^(300)-PDH蛋白含量,免疫荧光组织化学法检测脊髓背角中PDK4与星形胶质细胞标志物胶质纤维酸性蛋白(GFAP)共表达情况。结果与首次给药前比较,第1-5天第1次给药后1 h M组和MD组TFL明显延长(P<0.05)。与C组比较,M组第1-5天第1次给药后1 h TFL明显延长(P<0.05),脊髓背角PDK4、p-S^(300)-PDH、p-S^(293)-PDH蛋白含量明显升高(P<0.05);MD组第1-5天第1次给药后1 h TFL明显延长(P<0.05),脊髓背角p-S^(293)-PDH蛋白含量明显升高(P<0.05)。与M组比较,D组第1-5天第1次给药后1 h TFL明显缩短(P<0.05);MD组第3-5天第1次给药后1 h TFL明显延长(P<0.05);D组和MD组PDK4、p-S^(293)-PDH、p-S^(300)-PDH蛋白含量明显降低(P<0.05)。与D组比较,MD组第1-5天第1次给药后1 h TFL明显延长(P<0.05),脊髓背角p-S^(293)-PDH蛋白含量明显升高(P<0.05)。PDK4蛋白在脊髓背角细胞中表达,并与星形胶质细胞标志物GFAP共表达。结论吗啡耐受小鼠的脊髓背角星形胶质细胞中PDK4蛋白含量升高,PDK4抑制剂DCA抑制了下游p-S^(293)-PDH和p-S^(300)-PDH蛋白的表达。展开更多
文摘Several metabolic gene expressions are regulated in concert with muscle glycogen status. We hypothesized that intermittent exercise performed at high but sub-maximal intensities with long recovery periods would induce a low glycogen state that would stimul- ate peroxisome proliferator-activated receptor-γ coa- ctivator-1α (PGC1-α) and pyruvate dehydrogenase kinase-4 (PDK-4) gene expression in muscle. Nine young human subjects performed two intermittent exercise sessions. One session consisted of 60 s cycling bouts at VO2max (IE100%), and the other session consisted of 75 s cycling bouts at 80% VO2max (IE80%). Twelve bouts of exercise were completed in both sessions with a 4 min rest between each bout. Muscle specimens were obtained at pre-exercise and immediately, 1.5 h and 3 h post-exercise. Muscle gly- cogen was significantly decreased after both sessions (IE100%, 94.1 ± 5.8 to 38.7 ± 5.5 mmol/kg w.w.;IE80%, 94.6 ± 9.1 to 53.3 ± 4.8 mmol/kg w.w.;both P α and PDK- 4 mRNA expression were significantly increased after exercise in both IE100% and IE80% (PGC-1α: ~3.7 and ~2.9-fold, respectively;PDK-4: ~11.1 and ~3.5-fold, respectively;all P 100% than in IE80% (P a and PDK-4 mRNA expression, suggesting that increasing exercise intensity contributes to muscle glycogen depletion and PDK-4 mRNA expression in human skeletal muscle.
文摘目的探讨脊髓背角星形胶质细胞中丙酮酸脱氢酶激酶4(PDK4)在吗啡耐受小鼠中的作用。方法SPF级雄性C57b1/6小鼠24只,6~8周龄,体重22~25 g。采用随机数字表法分为四组:生理盐水组(C组)、吗啡组(M组)、PDK4抑制剂二氯乙酸(DCA)组(D组)和吗啡+DCA组四组(MD组),每组6只。C组鞘内注射生理盐水10μl,每日2次,连续5 d;M组、D组和MD组分别鞘内注射含吗啡10μg、DCA 200μg和吗啡10μg+DCA 200μg的生理盐水10μl,每日2次,连续5 d。首次给药前和每日第1次给药后1 h检测小鼠甩尾潜伏期(TFL)。最后一次给药后处死小鼠,采用Western blot法检测脊髓背角中PDK4、磷酸化丙酮酸脱氢酶(p-PDH)、p-S^(293)-PDH、p-S^(300)-PDH蛋白含量,免疫荧光组织化学法检测脊髓背角中PDK4与星形胶质细胞标志物胶质纤维酸性蛋白(GFAP)共表达情况。结果与首次给药前比较,第1-5天第1次给药后1 h M组和MD组TFL明显延长(P<0.05)。与C组比较,M组第1-5天第1次给药后1 h TFL明显延长(P<0.05),脊髓背角PDK4、p-S^(300)-PDH、p-S^(293)-PDH蛋白含量明显升高(P<0.05);MD组第1-5天第1次给药后1 h TFL明显延长(P<0.05),脊髓背角p-S^(293)-PDH蛋白含量明显升高(P<0.05)。与M组比较,D组第1-5天第1次给药后1 h TFL明显缩短(P<0.05);MD组第3-5天第1次给药后1 h TFL明显延长(P<0.05);D组和MD组PDK4、p-S^(293)-PDH、p-S^(300)-PDH蛋白含量明显降低(P<0.05)。与D组比较,MD组第1-5天第1次给药后1 h TFL明显延长(P<0.05),脊髓背角p-S^(293)-PDH蛋白含量明显升高(P<0.05)。PDK4蛋白在脊髓背角细胞中表达,并与星形胶质细胞标志物GFAP共表达。结论吗啡耐受小鼠的脊髓背角星形胶质细胞中PDK4蛋白含量升高,PDK4抑制剂DCA抑制了下游p-S^(293)-PDH和p-S^(300)-PDH蛋白的表达。
