CO2气腹对正常大鼠脑组织S-100、NSE mRNA和蛋白水平的影响

目的研究在不同CO2气腹压及不同时段下大鼠血和脑脊液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,RT-PCR技术和免疫组织化学的方法观察脑组织中S-100、NSEmRNA和蛋白水平表达的变化。结果CO2气腹后,A5-1组与对照组比较,海马区胶质细胞和神经元细胞均无明显变化,A10-2组胶质细胞和神经元细胞在光镜下呈轻度水样变,电镜下线粒体轻度肿胀;血液和脑脊液中S-100、NSE在CO2气腹压力后逐渐增加,但差异无统计学意义(P>0.05),S-100和NSE在血液和脑脊液中水平变化分别呈正相关(P<0.05);S-100、NSEmRNA在CO2气腹后逐渐增加,但与对照组比较差异无统计学意(P>0.05);脑组织S-100、NSE表达阳性面积随气腹时间及压力增加而逐渐增加,在A5-1组和A10-2组与对照组比较差异无统计学意义(P>0.05)。结论在10mmHg2h范围,CO2气腹对正常大鼠中枢神经系统无损害,CO2气腹在这一范围内应用对中枢神经系统是安全的。

Changes of mRNA and protein levels of S-100 and neuron specific enolase in normal brain tissues of rats following pneumoperitoneun with carbon dioxide

Objective To evaluate the changes of S-100 protein (S-100) and neuron specific enolase (NSE) of blood and cerebrospinal fluid (CSF) at different time and pressure of carbon dioxide pneumoperitoneun (CO_2 PP) and explore the effect of CO_2 PP on central nervous system (CNS) in normal rats. Methods Model of CO_2 PP in Wistar rats was established to observe astrocytes and neurons in the hippocamps of brian tissue of rat by light microscope and electron microscope. The changes of S-100 and NSE in blood and CSF were measured by ELISA. The mRNA and protein expressions of S-100 and NSE were tested by RT-PCR and immunohistochemical assay. Results There was no histologic change of hippocampus astrocytes and neurons between A_ 5-1 group and control group after CO_2 PP. The astrocytes and neurons in A_ 10-2 group showed local hydropic degeneration under light microscope and mitochondrion swelling under electron microscope. The levels of S-100 and NSE were increased in both blood and CSF samples in A_ 5-1 group and A_ 10-2 group after CO_2 PP, without statistical difference ( P >0.05). The levels of S-100 and NSE in the blood were found to be positively correlated with those in the CSF ( P <0.05). The mRNA expressions of S-100 and NSE increased progressively in A_ 5-1 group and A_ 10-2 group, without statistical difference compared with control group ( P >0.05). The positive expression areas of S-100 and NSE in the brain tissue increased progressively with increase of pressure and longer time of CO_2 PP, with insignificantly statistical difference compared with control group at points of A_ 5-1 and A_ 10-2 ( P >0.05). Conclusion CO_2 PP at pressure of 10 mm Hg and within 2 hours is safe to CNS in rats.