南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (08): 1192-1199.doi: 10.12122/j.issn.1673-4254.2020.08.18

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有氧运动联合HWTX-I调控II相解毒酶对阻塞性黄疸致中枢神经系统损伤的影响

梁 微，陈嘉勤，陈 伟   

  • 出版日期:2020-08-20 发布日期:2020-08-20
  • 基金资助:

Aerobic exercise combined with huwentoxin- I upregulates phase-II detoxification enzymes to alleviate obstructive jaundice-induced central nervous system injury in mice

  

  • Online:2020-08-20 Published:2020-08-20

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摘要： 目的 探讨有氧运动联合HWTX-I介导Keap1-Nrf2-ARE途径对II相解毒酶HO-1和NQO1的影响，以及在阻塞性黄疸小鼠中枢神经系统损伤中的保护作用。方法 50只雄性KM小鼠随机分为5组，10只/组：空白组（GO），模型组（M），有氧运动组（T），HWTX-I组（H），有氧运动联合HWTX-I组（TH）。M组、T组、H组、TH组小鼠均采用手术线直角悬挂阻断胆总管72 h构建阻塞性黄疸模型。通过有氧运动及尾静脉注射HWTX-I（0.05 µg/g）进行干预，采用一般行为学观察及Clark神经功能评分、ELISA酶联免疫、脑组织nissl染色、海马组织Western blot检测、mRNA检测及肝组织mRNA表达谱测序分析，探讨有氧运动及HWTX-I对阻塞性黄疸小鼠脑组织中枢神经损伤的改善效果。结果 M组胆总管悬挂72 h后出现明显黄疸症状，皮肤局灶黄染面积增大，尿液变黄及粪便呈浅灰色，Clark神经功能评分显著升高（P<0.01）；与M相比，T组、H组、TH组血清肝功能指标ALT、AST、TBIL和TBA含量水平下降（P<0.01），海马组织5-HT、BDNF含量上升，S100B、NSE含量下降（P<0.01），上述除肝功能指标外，有氧运动与HWTX-I间均存在协同效应。镜下显示，M组脑组织皮质神经细胞尼氏小体大量丢失，有明显空泡、深染，大量核固缩、核坏死。T组、H组针对上述症状明显改善，TH组干预效果最佳。相比M组，T组、H组、TH组脑组织中Nrf2、HO-1、NQO1 mRNA表达及蛋白表达水平显著升高（P<0.01，P<0.05），且有氧运动与HWTX-I间均存在协同效应。Illumina高通量测序筛选肝组织神经相关因子及关联分析显示，差异表达因子主要富集于神经活性配体-受体相互作用、GABA能突触、多巴胺能突触、突触小泡循环及轴突指导等神经调控通路，涉及组织细胞神经细胞信号转导、凋亡抑制、免疫反应、毒性等，有氧运动与HWTX-I可显著增加神经元损伤修复、增殖等相关信号通路的富集。且有氧运动与HWTX-I间均存在协同效应。结论 7周有氧运动或HWTX-I尾静脉注射可通过Keap1-Nrf2-ARE途径上调II相解毒酶HO-1和NQO1的表达，增强机体阻塞性黄疸后脑组织中枢神经的保护作用，且有氧运动与HWTX-I联合应用效果最佳。

Abstract: Objective To explore the effects of aerobic exercise combined with huwentoxin-I (HWTX-I)-mediated Keap1-Nrf2-ARE pathway on phase II detoxification enzymes HO-1 and NQO1 and their protective effects against obstructive jaundice (OJ)-induced central nervous system injury in mice. Methods 50 male KM mice were randomly divided into blank group (GO), model group (M), aerobic exercise group (T), HWTX-I group (H), and aerobic exercise combined with HWTX-I group (TH). Mouse models of OJ were established with surgical suture for 72 h in the mice in all the groups except for the blank control group. The mice received interventions by aerobic exercise and tail vein injection of HWTX-I (0.05 µg/g) and were assessed by behavioral observation, Clark’s neurological function scores, enzyme-linked immunosorbent assay (ELISA), brain tissue Nissl staining, hippocampal tissue Western blotting, and liver tissue mRNA expression profiling and sequencing. Results The mice in group M had obvious jaundice symptoms after the operation with significantly increased Clark’s neurological score (P<0.01). Compared with those in group M, the mice in group T, group H, and group TH showed significantly decreased serum levels of ALT, AST, TBIL, and TBA (P<0.01) with increased contents of 5-HT and BDNF and decreased contents of S100B and NSE in the hippocampus (P<0.01). Synergistic effects between aerobic exercise and HWTX-I were noted on the above parameters except for the liver function indicators. Interventions with aerobic exercise and HWTX-I, alone or in combination, obviously lessened pathologies in the brain tissue induced by OJ, and the combined treatment produced the strongest effect. The treatment also increased the expression levels of Nrf2, HO-1, and NQO1 mRNA and protein in brain tissues (P<0.01 or 0.05) with a synergistic effect between aerobic exercise and HWTX-I. Illumina high-throughput sequencing showed that the differentially expressed factors participated mainly in such neural regulatory pathways as neuroactive ligand-receptor interaction, GABAergic synapses, dopaminergic synapses, synaptic vesicle circulation, and axon guidance, involving tissue cell neuronal signal transduction, apoptosis inhibition, immune response, and toxicity. Aerobic exercise and HWTX-I synergistically increased the accumulation of the signal pathways related with neuron damage repair and proliferation. Conclusions Aerobic exercise combined with HWTX-I can up-regulate the expression of phase II detoxification enzymes HO-1 and NQO1 through the Keap1-Nrf2-ARE pathway to protect the central nervous system against OJ-induced damage in mice.