槲皮素对脊髓损伤大鼠神经元自噬及PI3K/Akt通路的影响  

Effects of Quercetin on Neuronal Autophagy and PI3K/Akt Pathway in Rats with Spinal Cord Injury

作　　者：聂晶鑫[1] 尚静波[1] 孟庆涛[1] Nie Jingxin;Shang Jingbo;Meng Qingtao(Dalian Third People′s Hospital,Liaoning,Dalian 116031,China)

机构地区：[1]辽宁省大连市第三人民医院,辽宁大连116031

出　　处：《中国中医急症》2020年第8期1415-1419,1439,共6页Journal of Emergency in Traditional Chinese Medicine

基　　金：辽宁省科学技术计划项目(2018225043)。

摘　　要：目的探讨槲皮素对脊髓损伤大鼠神经元自噬及PI3K/Akt通路的影响。方法SD大鼠100只随机分成5组:对照组、模型组、米诺环素组(90 mg/kg)、槲皮素低剂量组(50 mg/kg)、槲皮素高剂量组(100 mg/kg)。模型组、米诺环素组和槲皮素低、高剂量组建立脊髓损伤模型。建模成功后,米诺环素组和槲皮素低、高剂量组给予相应药物灌胃,对照组、模型组给予等体积的生理盐水,持续4周。给药结束后,进行脊髓损伤的行为学评分(BBB)、机械痛阈(PWT)、热刺激潜伏期(PWTL)的测定。吖啶橙染色观察大鼠海马神经元自噬水平;RTPCR法及免疫组织化学法测定大鼠海马组织中PI3K、Akt mRNA和蛋白表达水平,酶联免疫吸附法测定大鼠海马组织IL-4、IL-6、TNF-α蛋白表达水平。结果与对照组比较,模型组PWT、PWTL、BBB评分水平、神经元自噬小体水平、PI3K、Akt m RNA和蛋白表达水平明显降低(P<0.05),IL-4、IL-6、TNF-α蛋白表达水平明显升高(P<0.05)。与模型组比较,米诺环素组和槲皮素低、高剂量组PWT、PWTL、BBB评分水平、神经元自噬小体水平、PI3K、Akt mRNA和蛋白表达水平明显升高(P<0.05),IL-4、IL-6、TNF-α蛋白表达水平明显降低(P<0.05)。与米诺环素组比较,槲皮素低剂量组PWT、PWTL、BBB评分水平、神经元自噬小体水平、PI3K、Akt mRNA和蛋白表达水平明显降低(P<0.05),IL-4、IL-6、TNF-α蛋白表达水平明显升高(P<0.05);槲皮素高剂量组PWT、PWTL、BBB评分水平、神经元自噬小体水平、PI3K、Akt mRNA和蛋白表达水平、IL-4、IL-6、TNF-α蛋白表达水平相近(P>0.05)。结论槲皮素能明显增加脊髓损伤大鼠神经元自噬水平进而减轻脊髓神经元损伤程度,其机制可能与槲皮素通过激活PI3K/Akt信号通路诱导的大鼠神经元自噬来减弱脊髓损伤有关。Objective:To investigate the effects of Quercetin on neuronal autophagy and PI3 K/Akt pathway in rats with spinal cord injury.Methods:100 clean SD rats were randomly divided into five groups:the control group,the model group,Minocycline group(90 mg/kg),quercetin low dose group(50 mg/kg)and quercetin high dose group(100 mg/kg).The rats in the model group,Minocycline group and quercetin low and high dose group were used to establish spinal cord injury model.After successful modeling,Minocycline group,quercetin low-dose group and high-dose group were given corresponding drugs by gavage for 4 weeks,while the control group and the model group were given equal volume of physiological saline.After the experiment,BBB,PWT and PWTL of spinal cord injury were measured.Autophagy of hippocampal neurons was observed by acridine orange staining;PI3 K,Akt m RNA and protein expression in hippocampus were measured by RT-PCR and immunohistochemistry,and IL-4,IL-6 and TNF-αprotein expression in hippocampus were measured by enzyme-linked immunosorbent assay.Results:Compared with the control group,scores of PWT,PWTL and BBB,neuron autophagosomes and mRNA and protein expression levels of PI3 K and Akt in the model group were significantly decreased(P<0.05),and IL-4,IL-6,and TNF-αwere significantly increased(P<0.05).Compared with the model group,the scores of PWT,PWTL and BBB,neuron autophagosomes and m RNA and protein expression levels of PI3 K and Akt in Minocycline group and quercetin low and high dose groups were significantly increased(P<0.05),and the protein expression levels of IL-4,IL-6 and TNF-αwere significantly decreased(P<0.05).Compared with Minocycline group,the scores of PWT,PWTL and BBB,neuron autophagosomes and m RNA and protein expression levels of PI3 K and Akt in quercetin low dose group were significantly decreased(P<0.05),and the protein expression levels of IL-4,IL-6 and TNF-αwere significantly increased(P<0.05);those in high dose group were similar to those in Minocycline group(P>0.05).Conclusion:Quercetin can

关 键 词：槲皮素 脊髓损伤 神经元自噬 PI3K/AKT通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]