CO_2气腹对缺氧缺血性脑损伤大鼠脑组织S-100、NSE蛋白水平的影响

Effects of Carbon Dioxide Pneumoperitoneum on Protein Level of S-100 and NSE in Brain Tissues of Hypoxic-ischemic Brain Damage Rats

目的研究在不同CO2气腹压及不同时段下缺氧缺血性脑损伤大鼠血和脑脊液S-100蛋白(S-100)、神经元特异性烯醇化酶(NSE)水平的变化对中枢神经系统的影响.方法Wistar大鼠建立缺氧缺血性脑损伤模型和气腹模型,光镜和电镜观察大鼠脑组织胶质细胞和神经元细胞组织学变化;抗体夹心法检测血液和脑脊液中S-100、NSE水平变化,免疫组织化学的方法观察脑组织中S-100、NSE表达的变化.结果(1)CO2气腹后,B与对照组比较,海马区胶质细胞和神经元细胞均呈水样变,无核坏死,电镜下线粒体、内质网中度水肿;B10-2组胶质细胞和神经元细胞在光镜下呈弥漫性水样变,部分胶质细胞核浓缩和细胞皱缩,神经元细胞胞浆酸性细胞增多,电镜下胶质细胞水样变,线粒体肿胀、脊断裂,胶质细胞内髓鞘样结构形成,神经元细胞线粒体、内质网肿胀变性,核膜破裂;(2)B与对照组比较,血液和脑脊液中S-100、NSE水平均明显增高,差异有显著性(P<0.05);B5-1组与B组比较水平均有增加,差异无显著性,在B10-2组差异有显著性意义(P<0.05);(3)CO2气腹后脑组织S-100、NSE表达阳性面积随时间及压力增加而逐渐增加,B组与对照组或B10-2组与B组比较均有显著性差异(P<0.05).结论在10 mmHg、2 h范围,CO2气腹对缺氧缺血性脑损伤大鼠中枢神经系统有损害,应尽可能缩短气腹时间及采取相应的调控措施.

Objective To evaluate the changes of S- 100 protein （S- 100） and neuron specific enolase （NSE） in the blood and cerebrospinal fluid （CSF） at different of time and pressure of carbon dioxide pneumoperitoneum （CO2 PP） and explore the effect of CO2 PP on central nervous system （CNS） in hypoxic -ischemic brain damage rats. Methods Models of CO2 PP and hypoxic - ischemic brain damage in Wistar rats were established to observe astrocytes and neurons in the hippocampus of brain tissue of rat by light microscope and electron microscope. The changes of S - 100 and NSE in blood and CSF were measured by ELISA. Immuno - histochemical assay was used to detect the changes of the expression of S - 100 and NSE at the protein level. Results （1） There were significantly histological difference between B group and control group. The astrocytes and neurons in the hippocampus of brain tissue of rat in B group showed hydropic degeneration, no nuclear necrosis under light microscope, mitochondrion swelling under electron microscope. Astrocytes in the hippocampus of the B10-2 group showed diffuse hydropic degeneration, nuclear pyknosis, and cell shrinkage under light microscope, mitochondrion and glycogen swelling, fragmentation of ridge myelinogenesis in the astrocytes. Neuron in the B10-2 group showed diffuse hydropic degeneration, cytoplasmic hyperosinophilia, local nuclear pyknosis and cell shrinkage under light microscope and nuclear membrane disruption under electron microscope. （2） The findings showed a significantly increased levels of S - 100 and NSE in the B group compared with the control group （P 〈 0. 05） or the B10-2 group compared with the B group （P 〈0.05）, respectively. （3） The positive areas of the expression of S - 100 and NSE in the brain tissue increased progressively with the increase of pressure and time of CO2 PP. There were significance between B group and control group （P 〈0. 05） or B10-2 group and B group （P 〈 0. 05 ） . Conclusion CO2 PP at pressure of 10 mmHg and within 2 hours does produce central nervous system injury in rat with hypoxic -ischemic brain damage. It is necessary to short PP time or to take control means.