鞘内注射罗哌卡因-葡萄糖液对大鼠脊髓蛋白质组的影响

Effect of injection of ropivacaine with glucose into subarachnoid on the proteome of rat spinal cord

目的研究鞘内注射混合了葡萄糖的罗哌卡因对大鼠行为学指标及脊髓蛋白质组的影响。方法 36只大鼠随机分为3组,经腰椎鞘内置管后,按组别给予0.5%罗哌卡因(R1组)、10%葡萄糖+等体积1%罗哌卡因配制成的0.5%罗哌卡因(R2组)及对照组(N组)。每次注入40μL,每间隔1.5 h 1次,共3次。首次注药前5 min及每次注药后10 min测一次血压;置管后1 h、末次注药后1 h及24 h,以BBB神经功能评分法及斜板试验记录运动恢复情况。最后一次评分结束后断头法处死大鼠,以导管尖端为中心切取5 mm脊髓组织提取蛋白并定量,采用双向凝胶电泳-飞行时间质谱技术鉴定差异蛋白质。结果 R2组在末次给药后收缩压较基础值明显下降(P<0.05),与同时点对照组比较差异有统计学意义(P<0.05);该组BBB评分及斜板实验结果较基础值也略有下降,但差异无统计学意义(P>0.05)。R1组无上述改变。与对照组相比,R2组中谷氨酰胺合成酶及醛糖还原酶的表达升高,NAD依赖的去乙酰化酶2表达下降。结论由葡萄糖稀释的0.5%罗哌卡因用于大鼠蛛网膜下腔,24 h后其脊髓蛋白质中与细胞能量代谢相关的酶ALDR及GS表达升高,与细胞增殖分化相关的SIRT-2表达下调。

Objective To investigate the effects of ropivacaine injected intrathecally on the proteome of rat spinal cord. Methods 36 SD rats were randomly divided into three groups：group N（control group）,group R1,and group R2.After intrathecal catheterizations,0.5% ropivacaine（group R1） or the mixture solution of 10% glucose and 1% ropivacaine（group R2） were injected through the PE catheters individually.Every solution was given for three times with interval of 1.5 h and 40 μL per time.Systolic blood pressures were measured 5 min before the first injection and 10 min after each injection.Functional examinations were done at 1 h after catheterization,1 h after the last injection and 24 h after the last injection.The rats were killed,and 5 mm spinal cord was dissected out.Proteomics of the spinal cord samples were obtained by high resolution two-dimensional gel electrophoresis and protein spots with differences between control group and experiment group were chosen and picked by auto spots picker. Results The SBP of the group R2 decreased significantly after the injection of local anesthetics（P〈0.05）.The functional examination results of the group R2 showed decreased trends,but with no significant difference（P〈0.05）.The results of the group R1 showed no change.MS analysis showed that in group R2,the SIRT2 decreased 24 h after the injection,while the ALDR and the GS increased simultaneity. Conclusion 0.5% ropicaine with 10% glucose administered intrathecally can affect the proteomics of rat spinal cord.As a result,after 24 hour,the ALDR and GS increased while the SIRT2 decreased.