异丙酚改善氯胺酮诱发新生大鼠脑损伤的蛋白组学研究

Research on proteomics in propofol alleviates the brain injury of new born rats receiving ketamine anesthesia

目的探讨异丙酚复合氯胺酮麻醉引起幼年大鼠海马神经元以及认知功能损害后海马组织蛋白组学的变化,揭示海马神经元变化的病理机制,为提高临床麻醉安全提供指导。方法选择出生后7 d幼鼠80只分为生理盐水组(N组),氯胺酮组(K组),异丙酚+氯胺酮组(P+K组)低、高剂量组模拟幼儿麻醉状态,并进行海马组织切片及海马蛋白提取,应用双向电泳和基质辅助激光解析/电离-飞行时间质谱(MALDI-TOF)进行蛋白组学差异研究。结果水迷宫实验提示P组和P+K组术后认知功能明显优于K组;在海马神经元凋亡方面,P+K低剂量组[(10.23±4.82)%]及高剂量组[(6.79±6.48)%]均较K组凋亡指数[(14.98±5.65)%]明显降低(P<0.05),但P+K低剂量组与高剂量组间比较无显著性差异。K组海马组织蛋白浓度[(1.78±0.28)μg/μl]较N组明显下调[(2.11±0.32)μg/μl,P<0.05]。P+K各组[(1.88±0.27)%,(1.93±0.21)%]与N组相比差异无统计学意义。K+P组在6 h检测海马组织蛋白,发现上调蛋白并鉴定为PD1A3、NDUFB10、HSPA8、ATP5JD、PSMA1,下调蛋白为PPIA、PKM2、GFAP、NSE、PPIA、PKM2、GFAP等,21 d后检测结果表达上调蛋白为FUBP3、PRDX5,下调蛋白为GAPDH、AKR1A1、VC、TUBULIN A1 B。结论本实验中蛋白质组学技术初步筛选了异丙酚及氯胺酮麻醉后海马组织蛋白的差异表达,并分析了其在认知功能中的作用,为预防和改善海马神经元变化后认知功能损害提供理论基础。

Objective To investigate the damages of hippocampal neuron and cognitive function and changes of hippocampal proteins in a rat model anesthesia with propofol combined with ketamine, providing guidance for improving clinical anesthesia safety. Methods 80 infant rats(7 days after birth) were divided into saline group(N group), ketamine group(K group), propofol+ketamine group(P+K group)(this group was divided into two subgroups as low-dose P+K subgroups and high-dose P+K subgroup). After anesthesia, the hippocampus tissues was taken and the proteins in tissue were extracted. Two-dimensional electrophoresis and matrix assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF) were employed for analysis of proteomics. Results The outcomes of water maze indicated the cognitive function in P group or P+K group were superior to K group; The apoptosis index of hippocampal neurons in K group was significantly higher than high-dose P+K subgroup or low-dose P+K group(Both P<0.05), however, there was no statistical difference between two P+K subgroups, The level of hippocampal proteins in K group was statistically down-regulated compared with N group(P<0.05), but miss statistical difference while compared with both P+K subgroups. The unregulated proteins were detected in K+P group at 6 h were PD1A3, NDUFB10, HSPA8, ATP5 JD,PSMA1 respectively, while down-regulated proteins included PPIA, PKM2, GFAP, NSE, PPIA, PKM2, GFAP. At 21 d, the upregulated proteins were FUBP3, PRDX5, while down-regulated proteins were GAPDH, AKR1A1, VCP, TUBULIN A1 B. Conclusion In this study, we screened the protein difference while used ketamine or ketamine plus propofol for new born rat anesthesia with proteomics techniques. The result could contribute to provide theoretical support for alleviating cognitive function damage