Zhang Hua, Lu Min, Qi Hong-Bo, Zhang Jian-Hua
Department of Obstetrics and Gynecology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Zhonghua Fu Chan Ke Za Zhi. 2008 May;43(5):356-60.
To evaluate the influence of up-regulation of glucose regulated protein 78 (GRP 78) induced by 2-deoxyglucose (2DG) on fetal rat cerebral neuron apoptosis following intrauterine distress and the unification of endoplasmic reticulum and mitochondrium.
(1) Fetal rat intrauterine distress model was established and rats were divided into normal group (N = 10), ischemia- reperfusion(IR) group (n = 40) and treatment group (n = 40, injection of 2DG into pregnant rats' abdomen after operation ). (2) Neuron apoptosis and the influence of 2DG on apoptosis was detected by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining. The expression of GRP78, caspase-9, -12, and cytoron C protein were detected by western blot technique.
(1) The number of TUNEL positive neuron in normal group was 4.3 +/- 1.8/mm2. The expression of GRP78, caspase-9, -12, cytoron C in cytoplasm were 0.012 +/- 0.003, 0.004 +/- 0.003, 0.006 +/- 0.002, 0.012 +/- 0.005, respectively. (2) The number of TUNEL positive neuron in the IR group were 43.6 +/- 11.4/mm2 (reperfusion 3 h), 64.4 +/- 9.3/mm2 (reperfusion 6 h), 74.2 +/- 12.1/mm2 (reperfusion 12 h), 97.3 +/- 8.9/ mm2 (reperfusion 24 h), respectively. They were significantly more than that in normal group (P < 0.05). The expression of GRP78 at corresponding times in IR group were 0.092 +/- 0.008 (reperfusion 3 h), 0.078 +/- 0.006 (reperfusion 6 h), 0.054 +/- 0.009 (reperfusion 12 h), 0.038 +/- 0.007 (reperfusion 24 h), respectively. The expression of cytoron C in cytoplasm at corresponding times in IR group were 0.040 +/- 0.006 (reperfusion 3 h), 0.076 +/- 0.009 (reperfusion 6 h), 0.108 +/- 0.005 (reperfusion 12 h), 0.089 +/- 0.008 (reperfusion 24 h), respectively. The expression of caspase-9 at corresponding times in IR group were 0.042 +/- 0.003 ( reperfusion 3 h), 0.086 +/- 0.007 (reperfusion 6 h), 0.142 +/- 0.006 (reperfusion 12 h), 0.112 +/- 0.009 (reperfusion 24 h), respectively. The expression of caspase-12 at corresponding times in IR group were 0.076 +/- 0.006 (reperfusion 3 h), 0.113 +/- 0.010 (reperfusion 6 h), 0.125 +/- 0.005 (reperfusion 12 h), 0.057 +/- 0.008 (reperfusion 24 h), respectively. They were significantly higher than that in normal group (P < 0.05). (3) The number of TUNEL positive neuron in the treatment group were 19.4 +/- 10.6/mm2 (reperfusion 3 h), 26.4 +/- 12.3 /mm2 (reperfusion 6 h), 39.3 +/- 13.3/mm2 (reperfusion 12 h), 49.3 +/- 13.6/mm2 (reperfusion 24 h), respectively. They were significantly lower than that in IR group, but more than that in normal group (P < 0.05). The expression of GRP78 at corresponding times in the treatment group were 0.158 +/- 0.012 (reperfusion 3 h), 0.175 +/- 0.005 (reperfusion 6 h), 0.125 +/- 0.013 (reperfusion 12 h), 0.079 +/- 0.004 (reperfusion 24 h), respectively. They were significantly higher than that in IR group and normal group (P < 0.05) . The expression of cytoron C in cytoplasm at corresponding times in IR group were 0.026 +/- 0.002 (reperfusion 3 h), 0.042 +/- 0.008 (reperfusion 6 h), 0.062 +/- 0.007 (reperfusion 12 h), 0.045 +/- 0.004 (reperfusion 24 h), respectively. The expression of caspase-9 at corresponding times in IR group were 0.033 +/- 0.002 (reperfusion 3 h), 0.063 +/- 0.005 (reperfusion 6 h), 0.092 +/- 0.005 (reperfusion 12 h), 0.068 +/- 0.008 (reperfusion 24 h), respectively. The expression of caspase-12 at corresponding times in IR group were 0.061 +/- 0.004 (reperfusion 3 h), 0.068 +/- 0.009 (reperfusion 6 h), 0.072 +/- 0.007 (reperfusion 12 h), 0.054 +/- 0.005 (reperfusion 24 h), respectively. They were significantly lower than that in IR group, but higher than that in normal group (P < 0.05).
Fetal rat cerebral neuron apoptosis following intrauterine distress is associated with the action of endoplasmic reticulum and mitochondrium. Up-regulation of GRP78 induced by 2DG counteracts primary cellular damage caused by endoplasmic reticulum stress. 2DG plays a protective role for fetal rat cerebral neuron following intrauterine distress.
评估2-脱氧葡萄糖（2DG）诱导的葡萄糖调节蛋白78（GRP 78）上调对宫内窘迫后胎鼠脑神经元凋亡及内质网与线粒体联合作用的影响。
（1）建立胎鼠宫内窘迫模型，将大鼠分为正常组（N = 10）、缺血再灌注（IR）组（n = 40）和治疗组（n = 40，术后向孕鼠腹腔注射2DG）。（2）采用末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记（TUNEL）染色检测神经元凋亡及2DG对凋亡的影响。采用蛋白质免疫印迹技术检测GRP78、半胱天冬酶-9、-12和细胞色素C蛋白的表达。
（1）正常组TUNEL阳性神经元数量为4.3±1.8/mm²。GRP78、半胱天冬酶-9、-12、细胞色素C在细胞质中的表达分别为0.012±0.003、0.004±0.003、0.006±0.002、0.012±0.005。（2）IR组TUNEL阳性神经元数量在再灌注3 h时为43.6±11.4/mm²，再灌注6 h时为64.4±9.3/mm²，再灌注12 h时为74.2±12.1/mm²，再灌注24 h时为97.3±8.9/mm²，均显著高于正常组（P < 0.05）。IR组相应时间GRP78的表达分别为0.092±0.008（再灌注3 h）、0.078±0.006（再灌注6 h）、0.054±0.009（再灌注12 h）、0.038±0.007（再灌注24 h）。IR组相应时间细胞质中细胞色素C的表达分别为0.040±0.006（再灌注3 h）、0.076±0.009（再灌注6 h）、0.108±0.005（再灌注12 h）、0.089±0.008（再灌注24 h）。IR组相应时间半胱天冬酶-9的表达分别为0.042±0.003（再灌注3 h）、0.086±0.007（再灌注6 h）、0.142±0.006（再灌注12 h）、0.112±0.009（再灌注24 h）。IR组相应时间半胱天冬酶-12的表达分别为0.076±0.006（再灌注3 h）、0.113±0.010（再灌注6 h）、0.125±0.005（再灌注12 h）、0.057±0.008（再灌注24 h），均显著高于正常组（P < 0.05）。（3）治疗组TUNEL阳性神经元数量在再灌注3 h时为19.4±10.6/mm²，再灌注6 h时为26.4±12.3/mm²，再灌注12 h时为39.3±13.3/mm²，再灌注24 h时为49.3±13.6/mm²，均显著低于IR组，但高于正常组（P < 0.05）。治疗组相应时间GRP78的表达分别为0.158±0.012（再灌注3 h）、0.175±0.005（再灌注6 h）、0.125±0.013（再灌注12 h）、0.079±0.004（再灌注24 h），均显著高于IR组和正常组（P < 0.05）。治疗组相应时间细胞质中细胞色素C的表达分别为0.026±0.002（再灌注3 h）、0.042±0.008（再灌注6 h）、0.062±0.007（再灌注12 h）、0.045±0.004（再灌注24 h）。治疗组相应时间半胱天冬酶-9的表达分别为0.033±0.002（再灌注3 h）、0.063±0.005（再灌注6 h）、0.092±0.005（再灌注12 h）、0.068±0.008（再灌注24 h）。治疗组相应时间半胱天冬酶-12的表达分别为0.061±0.004（再灌注3 h）、0.068±0.009（再灌注6 h）、0.072±0.007（再灌注12 h）、0.05
