利多卡因对老年大鼠术后认知功能障碍炎症反应的影响

发布时间：2018-07-06 06:44

本文选题：利多卡因 + 术后认知功能障碍　；参考：《中国人民解放军医学院》2014年博士论文

【摘要】：第一部分不同剂量利多卡因对老年大鼠POCD行为学影响目的：通过脾切除构建老年大鼠术后认知功能障碍（POCD）模型，探索老年大鼠POCD模型形成及维持时间，并以局部麻醉药利多卡因作为研究工具，，研究利多卡因对学习记忆功能损伤的改善作用，确定利多卡因改善POCD的最佳剂量。方法：将50只老年SD大鼠，使用CHISS统计学软件随机分成5组，组别分别为：对照组、POCD模型组、利多卡因1组（1mg/kg/h）、利多卡因2组（2mg/kg/h）、利多卡因3组（4mg/kg/h）,每组10只。进行连续5天的空间获得性实验后，第6天对各组老年SD大鼠进行模型构建和利多卡因干预。所有老年SD大鼠均在术后第1、3、7、14天利用Morris水迷宫对老年大鼠的空间记忆能力进行测试。结果：空间获得性实验会使老年大鼠的学习记忆能力随着时间的延长而逐渐提高（P 0.01）、并在第5天趋于稳定，且每天各组之间的没有明显差异。参考记忆的测试结果显示，与对照组相比，POCD模型组和利多卡因3组（4mg/kg/h）的参考记忆能力明显降低（P 0.05）;与POCD模型组相比，利多卡因1组（1mg/kg/h）和利多卡因3组（4mg/kg/h）的参考记忆能力明显增强（P 0.05）。工作记忆的测试结果显示，与对照组相比，POCD模型组、利多卡因1组（1mg/kg/h）术后第1、3天，利多卡因3组（4mg/kg/h）术后第1、3、7天的工作记忆能力有显著差异（P 0.05）;与POCD模型组比较，利多卡因1、2组（1、2mg/kg/h）的术后第1、3天和利多卡因3组（4mg/kg/h）的术后第1天的工作记忆能力有显著差异（P0.05）。结论：麻醉状态下脾切除能够成功建立POCD模型，且空间记忆能力的损害在术后第3天最明显，术后第7天基本恢复正常，而围术期尾静脉持续泵注1h的1mg/kg/h利多卡因对构建POCD模型后的老年大鼠空间记忆能力的改善作用最明显，被确定为最佳治疗剂量。第二部分最佳剂量利多卡因对POCD老年大鼠海马组织和外周血炎症反应的影响目的：通过分子生物学和免疫组织化学技术，观察最佳剂量利多卡因对POCD老年大鼠海马和外周血炎症反应的影响，探索中枢和外周炎症反应与海马星形胶质细胞激活和表达变化的机制。方法：确定量效关系和利多卡因的最佳剂量后，将72只老年SD大鼠，使用CHISS统计学软件随机分成3组，组别分别为：对照组、POCD模型组、利多卡因组（1mg/kg/h）。使用54只进行Morris水迷宫实验，每组18只，进行连续5天的空间获得性实验后，第6天对各组老年SD大鼠进行模型构建和利多卡因干预，术后第1、3、7天对老年大鼠的空间记忆能力进行测试。使用18只进行新物体识别实验，每组6只，术后第1天对老年大鼠的非空间记忆能力进行测试。所有老年大鼠均在术后第1、3、7天取材，采用ELISA的方法对老年大鼠海马组织和外周血浆炎症细胞因子进行检测，采用组织免疫荧光染色和蛋白免疫印迹的方法观察老年SD大鼠海马组织中星形胶质细胞激活和GFAP表达的情况。结果：确定利多卡因的最佳治疗剂量为1mg/kg/h后，不仅使用Morris水迷宫实验评价老年大鼠的空间记忆能力，还用新物体识别实验评价非空间记忆能力，其结果显示，与对照组相比，POCD模型组老年大鼠识别指数有显著差异（P 0.05）；与POCD模型组对比，利多卡因组（1mg/kg/h）老年大鼠新物体识别指数有显著差异（P 0.05）。测定炎症细胞因子结果显示，关于促炎因子，与对照组相比，POCD模型组术后第1天和第3天老年大鼠海马和血浆中的TNF-α、IL-1β和IL-6浓度分别有显著差异（P 0.05）；与POCD模型组相比，1mg/kg/h利多卡因干预后，术后第1天和第3天老年大鼠海马和血浆中的TNF-α、IL-1β和IL-6浓度分别有显著差异（P 0.05）。而抗炎因子IL-10，与对照组相比，POCD模型组在术后第1、3天的海马和血浆中的IL-10浓度分别有显著差异（P 0.05）；与POCD模型组相比，利多卡因组（1mg/kg/h）术后第3天海马和血浆中的IL-10浓度有显著差异（P 0.05）。组织免疫荧光染色法和蛋白免疫印迹法测定老年大鼠海马组织星形胶质细胞标志物GFAP的结果显示，与对照组相比，POCD模型组术后第1天和第3天海马组织中GFAP的表达有显著差异（P 0.05），且GFAP免疫荧光强度增加；与POCD模型组相比，利多卡因组（1mg/kg/h）术后第3天海马组织中GFAP的表达有显著差异（P 0.05），且GFAP免疫荧光强度显著减弱。结论：围术期尾静脉持续泵注1h的1mg/kg/h利多卡因对构建POCD模型后的老年大鼠空间记忆能力和非空间记忆能力均具有明显的改善作用，也能够很好地控制老年大鼠海马和血浆中炎症细胞因子（包括促炎因子和抗炎因子）的产生。应用1mg/kg/h利多卡因，能够明显减弱老年大鼠在术后第1天和第3天海马组织中星形胶质细胞的激活和星形胶质细胞特异性标志物GFAP的表达。
[Abstract]:Effects of different doses of lidocaine on the behavior of POCD in aged rats

Objective : To establish a model of cognitive impairment ( POCD ) in aged rats by splenic resection , to explore the formation and maintenance time of POCD in aged rats , and to study the effect of lidocaine on learning and memory impairment by using lidocaine as the research tool , and to determine the optimal dose of lidocaine for improving POCD .

Methods : 50 old SD rats were randomly divided into 5 groups using CHISS statistical software . The rats were randomly divided into five groups : control group , POCD model group , lidocaine group 1 group ( 1mg / kg / h ) , lidocaine group 2 ( 2mg / kg / h ) and lidocaine 3 group ( 4mg / kg / h ) . All the old SD rats were tested on the 1st , 3rd , 7th and 14th days after operation .

Results : The results showed that the learning and memory ability of aged rats gradually increased with time ( P 0.01 ) , and there was no significant difference between groups on Day 5 . The reference memory capacity of POCD model group and lidocaine group 3 group ( 4mg / kg / h ) was significantly lower than that in control group ( P 0.05 ) . Compared with control group , the reference memory ability of the group of lidocaine group 1 ( 1mg / kg / h ) and lidocaine group 3 ( 4mg / kg / h ) was significantly increased ( P 0.05 ) .

The results of working memory showed that compared with the control group , there was a significant difference in the working memory ability of the POCD model group , the lidocaine group 1 group ( 1 mg / kg / h ) on the first , 3 and 7 days after the operation ( P 0.05 ) . Compared with the POCD model group , there was a significant difference in the working memory capacity ( P0.05 ) in the first , 3 and 3 days after the operation of the lidocaine 1,2 group ( 1 , 2 mg / kg / h ) and the first day after the operation of the lidocaine group 3 ( 4mg / kg / h ) .

Conclusion : The POCD model can be successfully established under the anesthesia state , and the damage of spatial memory ability is the most obvious at the third day of operation , the 7th day after the operation is basically normal , and the effect of 1 mg / kg / h lidocaine on the space memory ability of the old rats after the perioperative tail vein continuous pumping is the most obvious and is determined as the optimal therapeutic dose .

Effect of the second part of optimal dose lidocaine on the inflammatory response of hippocampus and peripheral blood in aged rats with POCD

Objective : To investigate the effects of lidocaine on inflammatory responses in hippocampus and peripheral blood of elderly rats with POCD by molecular biology and immunohistochemical technique .

Methods : 72 old SD rats were randomly divided into three groups : control group , POCD model group , lidocaine group ( 1mg / kg / h ) .
Compared with POCD model group , there was a significant difference in the identification index of new objects in the aged rats ( 1mg / kg / h ) ( P 0.05 ) .

The results showed that the levels of TNF - 伪 , IL - 1尾 and IL - 6 in the hippocampus and plasma of the aged rats were significantly higher than those in the control group ( P 0.05 ) .
Compared with the POCD model group , the levels of TNF - 伪 , IL - 1尾 and IL - 6 in the hippocampus and plasma of the aged rats were significantly different after the intervention of 1 mg / kg / h lidocaine ( P 0.05 ) . Compared with the control group , the levels of IL - 10 in the hippocampus and plasma of POCD model group were significantly higher than those in control group ( P 0.05 ) .
Compared with POCD model group , the concentration of IL - 10 in hippocampus and plasma of lidocaine group ( 1 mg / kg / h ) was significantly different ( P 0.05 ) .

The expression of GFAP in hippocampus of aged rats was significantly higher than that in control group ( P 0.05 ) , and GFAP immunofluoresence intensity increased .
Conclusion : The effect of 1 mg / kg / h lidocaine on the spatial memory and non - spatial memory in the hippocampus and plasma of aged rats was significantly improved compared with the POCD model group ( P 0.05 ) . Conclusion : The effect of 1 mg / kg / h lidocaine on the spatial memory and non - spatial memory of the aged rats after the operation of the POCD model can be well controlled . The activation of astrocytes in the hippocampus and hippocampus of the aged rats and the expression of GFAP by astrocytes in the hippocampus and hippocampus of the aged rats can be significantly reduced .
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R614

【参考文献】