艾司氯胺酮调节PI3K/Akt/mTOR信号通路对抑郁症大鼠认知障碍的影响

Effect of Esketamine on Cognitive Impairment in Depressed Rats via Regulation of the PI3K/Akt/mTOR Signaling Pathway

目的:探讨艾司氯胺酮(ISLAT)调节PI3K/Akt/mTOR信号通路对抑郁症大鼠认知障碍的影响。方法:将大鼠随机分为CK组、Model组、ISLAT低剂量(ISLAT-L)组、ISLAT高剂量(ISLAT-H)组、ISLAT-H+LY294002(PI3K通路抑制剂)组,各组大鼠腹腔注射相应药物,日一次,持续3周。采用旷场实验、糖水偏好实验、强迫游泳实验评价大鼠抑郁行为和认知功能;ELISA法检测血清中TNF-α、IL-10、IL-6水平和海马组织中脑源性神经营养因子(BDNF)、去甲肾上腺素(NE)、多巴胺(DA)、5羟色胺(5-HT);HE染色观察大鼠海马组织损伤;TUNEL检测大鼠海马神经元凋亡;Western blot检测大鼠海马组织Cleaved-caspase-3、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达。结果:与CK组相比,Model组大鼠海马神经元形态有明显损伤,站立次数和蔗糖水饮用量、血清IL-10水平、海马组织中BDNF、NE、DA、5-HT水平、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达水平显著降低,游泳不动时间、血清TNF-α和IL-6水平、神经元凋亡率、cleaved-caspase-3蛋白表达水平显著升高(P<0.05);与Model组相比,ISLAT-L组和ISLAT-H组大鼠海马神经元形态有明显改善,站立次数和蔗糖水饮用量、血清IL-10水平、海马组织中BDNF、NE、DA、5-HT水平、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达水平显著升高,游泳不动时间、血清TNF-α和IL-6水平、神经元凋亡率、cleaved-caspase-3蛋白表达水平显著降低(P<0.05);LY294002可减轻ISLAT对抑郁症大鼠认知的改善作用(P<0.05)。结论:ISLAT通过激活PI3K/Akt/mTOR信号通路可减轻抑郁症大鼠炎症,降低神经组织损伤和神经元凋亡,改善大鼠抑郁症症状。

Objective:To investigate the effect of esketamine(ISLAT)on cognitive impairment in rats with depression by regulating the PI3K/Akt/mTOR signaling pathway.Methods:Rats were randomly separated into CK group,Model group,ISLAT low-dose(ISLAT-L)group,ISLAT high-dose(ISLAT-H)group,and ISLAT-H+LY294002(PI3K pathway inhibitor)group,all rats intraperitoneally injected with the corresponding drug for once a day for 3 weeks.The open field experiment,sugar water preference experiment,and forced swimming experiment were applied to evaluate the depressive behavior and cognitive function of rats.ELISA method was applied to detect the levels of TNF-α,IL-10,and IL-6 in serum,and brain-derived neurotrophic factor(BDNF),norepinephrine(NE),dopamine(DA),and serotonin(5-HT)in hippocampal tissue.HE staining was applied to observe hippocampal tissue damage in rats.TUNEL was applied to detect apoptosis of hippocampal neurons in rats.Western blot was applied to detect the expression of Cleaved-caspase-3,p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR proteins in rat hippocampal tissue.Results:Compared with the CK group,the model group showed great damage to the morphology of hippocampal neurons in rats,the number of standing times,sucrose water consumption,serum IL-10 level,BDNF,NE,DA,5-HT levels in hippocampal tissue,p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR protein expression levels were greatly reduced,the swimming immobility time,serum TNF-αand IL-6 levels,neuronal apoptosis rate,and Cleaved-caspase-3 protein expression level were greatly increased(P<0.05).Compared with the Model group,the morphology of hippocampal neurons in the ISLAT-L and ISLAT-H groups showed great improvement,the number of standing times,sucrose water consumption,serum IL-10 level,BDNF,NE,DA,5-HT levels in hippocampal tissue,p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR protein expression levels were greatly increased,the swimming immobility time,serum TNF-αand IL-6 levels,neuronal apoptosis rate,and Cleaved-caspase-3 protein expression level were greatly reduced(P<0.05).LY294002 was able to alleviate the cognitive improvement effect of ISLAT on depression rats(P<0.05).Conclusion:ISLAT can alleviate inflammation,reduce nerve tissue damage and neuronal apoptosis,and improve depression symptoms in rats by activating the PI3K/Akt/mTOR signaling pathway.