利用高分辨氢质子磁共振波谱技术探讨丙酮酸钠对反复严重低血糖新生大鼠枕叶皮层的保护作用及其机制  

Sodium pyruvate protect occipital cortex of rats with repetitive and severe neonatal hypoglycemia detected by high resolution 1H nuclear magnetic resonance spectroscopy

作　　者：刘锟[1] 付玉川[1] 杜美美[1] 叶信健[1] 黄小燕[1] 周永进[1] 赵良才[2] 高红昌[2] 严志汉[1] Liu Kun Fu Yuchuan Do Meimei Ye Xinjian Huang Xiaoyan Zhou Yongjin Zhao Liangcai Gao Hongchang Yan Zhihan.(Department of Radiology, the Second Affiliated Hospital and Yuying Children＇s Hospital of Wenzhou Medical University, Wenzhou 325000, Chin School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou 325035, China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院放射科,325000 [2]温州医科大学药学院,325035

出　　处：《中华围产医学杂志》2017年第3期228-233,共6页Chinese Journal of Perinatal Medicine

基　　金：国家自然科学基金（81400863）;浙江省医药卫生科技项目（2014KYB159、2015KYB250）;浙江省温州市科技计划项目（Y20130175、Y20140028）

摘　　要：目的应用高分辨氢质子磁共振波谱技术研究反复严重低血糖新生大鼠经丙酮酸钠干预治疗对脑损伤的保护作用及其机制。方法36只2日龄Sprague—Dawley大鼠随机分为低血糖组和干预组，每组18只。2组大鼠2、4、6日龄时腹腔注射胰岛素40U／kg诱导严重低血糖（血糖≤1．4mmol／L），低血糖组注射胰岛素后2．5h腹腔注射50％葡萄糖溶液2ml／kg终止低血糖，干预组则在注射同等量葡萄糖的同时注射丙酮酸钠溶液2．5ml／kg（500mg／kg）。7日龄时，每组选取6只大鼠，取枕叶皮层行原位末端转移酶标记技术染色以观察脑细胞的损伤情况；另外12只大鼠的枕叶皮层行离体高分辨氢质子磁共振波谱检查以观察代谢物变化情况。采用两独立样本t检验进行统计学分析。结果2、4和6日龄时，低血糖组及干预组新生大鼠在给予胰岛素2．5h后均达到严重低血糖水平。与低血糖组相比，干预组枕叶皮层内损伤神经元个数减少[（45±5）与（113±12）个，t=0．782，P=0．013]，损伤指数下降（0．15±0．03与0．36±0．06，t=1．143，P=0．020）。相较低血糖组，干预组大鼠枕叶皮层中牛磺酸f（13．31±2．06）与（18．44±3．86）mol／kg，t=8．231]、谷氨酰胺[（1．50±0．24）与（2．02±0．40）mol／kg，t=3．137]、谷氨酸[（7．04±0．95）与（9．40±1．73）mol／kg，t=6．449]、天冬氨酸[（1．51±0．28）与（2．15±0．58）mol／kg，t=2．561]和肌酸[（6．37±O．99）与（8．46±1．77）mol／kg，t=4．226]的浓度显著下降，差异均有统计学意义（P’值均〈O．017）。结论反复严重新生期低血糖在给予葡萄糖终止低血糖的同时提供丙酮酸钠可通过调节兴奋性氨基酸类神经递质、能量代谢及其他代谢通路而减轻低血糖所致的枕叶损伤，发挥脑保护作用。Objectives To investigate the occipital cortex metabolite alterations in repetitive and severe neonatal hypoglycemia rats treated with sodium pyruvate and to reveal the protective role of sodium pyruvate using high resolution ~H nuclear magnetic resonance spectroscopy. Methods Thirty-six 2-dayold Sprague-Dawley rats were randomly divided into hypoglycemia group and pyruvate group with 18 rats in each group. Rats in both groups received intraperitoneal injections of insulin （40 U/kg body weight） at 2, 4 and 6 days of age to induce severe hypoglycemia （blood glucose value ≤ 1.4 mmol/L）. In the hypoglycemia group, 2.5 hours after insulin injection, intraperitoneal injection of 50% glucose （2 ml/kg） was administered to terminate hypoglycemia, while in the pyruvate group, 50% glucose （2 ml/kg） and sodium pyruvate solution 2.5 ml/kg （500 mg/kg） were injected. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay was used to observe the status of injured neurons in six neonatal rats, and metabolite changes in occipital cortex of the other 12 rats were detected by ＇H nuclear magnetic resonance spectroscopy. The difference between the two groups was compared by independent-samples t test. Results Neonatal rats of both groups reached severe hypoglycemia level 2.5 hours after insulin injection. Compared with hypoglycemia group, pyruvate group had fewer injured neurons （45 ±5 vs 113 ±12, t=-0.782, P=0.013） and lower injured index in the occipital cortex （0.15 ± 0.03 vs 0.36± 0.06, t= 1.143, P=0.020）. Pyruvate group showed significant decreases in the concentration of taurine [（13.31 ± 2.06） vs （ 18.44 ± 3.86） mol/kg, t=-8.231 ], glutamine[（1.50 ± 0.24） vs （2.02 ± 0.40） mol/kg, t=3.137], glutamate[（7.04±0.95） vs （9.40±1.73） mol/kg, t=-6.449], aspartate[（1.51 ±0.28） vs （2.15±0.58） mol/kg, t=2.561] and creatine [（6.37±0.99） vs （8.46± 1.77） mol/kg, t=4.226] in the occipital cortex （all P〈0.017）. Conclusions Si

关 键 词：低血糖症 丙酮酸盐类 枕叶 磁共振波谱学 动物 新生 

分 类 号：R722.1[医药卫生—儿科]