摘要
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。
Objective To investigate the role of pyruvate dehydrogenase kinase 4(PDK4)in spinal dorsal horn astrocytes in morphine-tolerant mice.Methods Twenty-four SPF male C57b1/6 mice,6-8 weeks old,weighing 22-25 g,were divided into four groups by random number table method:normal saline group(group C),morphine group(group M),dichloroacetic acid(DCA)group(group D)and morphine+DCA group(group MD),6 mice in each group.Group C was intrathecally injected with normal saline 10μl,twice a day for 5 consecutive days;groups M,D and MD were intrathecally injected with normal saline containing morphine 10μg,DCA 200μg and morphine 10μg+DCA 200μg,twice a day for 5 consecutive days.The tail-flick latency(TFL)of mice was detected before the first administration and 1 hour after the first daily administration.The mice were sacrificed after the last administration,and the protein contents of PDK4,phosphorylated pyruvate dehydrogenase(p-PDH),p-S^(293)-PDH,and p-S^(300)-PDH in the dorsal horn of the spinal cord were detected by Western blot,and immunofluorescence histochemistry was used to detect the co-expression of PDK4 and astrocyte marker glial fibrillary acidic protein(GFAP)in the dorsal horn of spinal cord.Results Compared with before the first administration,TFL was significantly prolonged in groups M and MD 1 hour after the first daily administration on day 1-5(P<0.05).Compared with group C,TFL was significantly prolonged 1 hour after the first daily administration on day 1-5(P<0.05),and the protein contents of PDK4,p-S^(300)-PDH,and p-S^(293)-PDH in the dorsal horn of spinal cord were significantly increased in group M(P<0.05);the TFL was significantly prolonged 1 hour after the first daily administration on day 1-5(P<0.05),and the protein content of p-S^(293)-PDH in the spinal dorsal horn was significantly increased in group MD(P<0.05).Compared with group M,the TFL was significantly shortened 1 hour after the first daily administration on day 1-5 in group D(P<0.05);the TFL was significantly prolonged 1 hour after the first da
作者
程中亮
马霞青
许涛
王爱忠
CHENG Zhong-liang;MA Xiaqing;XU Tao;WANG Aizhong(School of Fisheries and Life,Shanghai Ocean University,Shanghai 201306,China)
出处
《临床麻醉学杂志》
CAS
CSCD
北大核心
2022年第10期1087-1091,共5页
Journal of Clinical Anesthesiology
基金
国家自然科学基金(81772062)。
关键词
吗啡耐受
丙酮酸脱氢酶激酶4
丙酮酸脱氢酶
磷酸化
小鼠
Morphine tolerance
Pyruvate dehydrogenase kinase 4
Pyruvate dehydrogenase
Phosphorylation
Mice
