虾青素与氯化锂治疗慢性有机磷中毒小鼠认知障碍的协同作用

The synergetic effect of combined astaxanthin and lithium chloride treatment on cognitive dysfunc-tion of chronic omethoate poisoned mice

目的：探讨虾青素与氯化锂治疗慢性有机磷中毒小鼠认知障碍的协同作用。方法成年健康雄性昆明小鼠55只，采用随机数字法选8只作为对照组，其余47只连续氧乐果染毒4周建立慢性有机磷中毒认知障碍模型，将造模成功的40只小鼠随机分为模型组，虾青素治疗组，依达拉奉治疗组，氯化锂治疗组和虾青素＋氯化锂联合治疗组，每组8只。应用Morris水迷宫检测小鼠的学习记忆能力；ELISA法检测脑组织活性氧（ ROS）含量，化学比色法检测超氧化物歧化酶（ SOD）活性；HE染色观察小鼠海马组织形态学变化，免疫组织化学染色和Western blot检测p-PI3K，p-Akt，p-GSK3β与p-CREB在海马组织内的分布和表达水平。结果各组小鼠5 d平均逃避潜伏期差异具有统计学意义（F＝1662．147， P＜0．05），虾青素＋氯化锂组5 d平均逃避潜伏期用时明显低于模型组（均 P＜0．05），并均低于其他治疗组；与对照组相比，模型组小鼠海马神经元出现明显损伤，p-PI3K（0．032±0．008），p-Akt（0．030±0．006），p-GSK3β（0．028±0．007）与p-CREB（0．020±0．008）在海马区表达水平较对照组（0．087±0．007，0．084±0．009，0．097±0．002，0．076±0．012）明显降低（均P＜0．05）；虾青素＋氯化锂组小鼠海马神经元损伤较模型组有所减轻，p-PI3K（0．067±0．008），p-Akt（0．065±0．005），p-GSK3β（0．068±0．009）和p-CREB（0．062±0．008）在海马区表达水平较模型组显著升高（均P＜0．05）。结论虾青素与氯化锂联合治疗通过上调Akt／GSK3β／CREB信号通路的磷酸化水平，对慢性有机磷中毒致认知障碍起到神经保护作用。

Objective To investigate the synergetic effect of combined astaxanthin （ AST） and lith-ium chloride （ LiCl） treatment on cognitive dysfunction of chronic omethoate poisoned mice. Methods 8 mice were selected randomly as control group from 55 healthy adult male Kunming mice,and the rest were used to establish chronic organophosphate poisoning cognitive impairment models by injecting omethoate 5 mg/kg subcutaneously every day for 4 weeks. Totally 40 successfully established models were randomly divid-ed into model group,AST group,edaravone group,LiCl group and AST＋LiCl group with 8 in each. Morris wa-ter maze test was used to examine the learning and memory ability of mice. Contents of reactive oxygen spe-cies （ROS） in hippocampus were measured by enzyme-linked immunosorbent assay （ELISA）. Activity of superoxide dismutase （ SOD） in hippocampus was measured by colorimetric assay. Morphology of hippocam-pus area was observed by HE staining. The distribution and expression of p-PI3K,p-Akt,p-GSK3β and p-CREB were determined by immunohistochemical staining （ IHC staining） and Western blot. Results The average escape latency of 5 days in each group was statistically significant （F=1662.147, P〈0.05） . The av-erage escape latency of 5 days in AST＋LiCl group was significantly lower than that in model group （ all P〈0.05） and was lower than other treatment groups. Compared with the control group （0.087±0.007,0.084± 0.009,0.097±0.002,0.076±0.012）,the hippocampal neuronal injury in model group was serious,the expres-sions of p-PI3K （0.032±0.008）,p-Akt （0.03±0.006）,p-GSK3β （0.028±0.007） and p-CREB （0.020± 0.008） was significantly lower （ all P〈0.05） . The injuries of hippocampal neurons in AST＋LiCl group were slightly lighter than that in model group,and the expression of p-PI3K （0.067±0.008）,p-Akt （0.065± 0.005）,p-GSK3β （0.068±0.009） and p-CREB （0.062±0.008） in hippocampus was significantly higher than that in model group （ all P〈0.05） . Conclusion Combined AST and LiCL treatment exerts neuroprotec-tive effect on cognitive dysfunction induced by chronic organophosphate poisoning via up-regulating the ex-pression of Akt/GSK3β/CREB.