胆酸和去氧胆酸抗惊厥作用的血清代谢组学研究  

Serum metabolomics study of the anticonvulsant effects of cholic acid and deoxycholic acid

作　　者：周俊发 乔婷婷 张志宏[2] 叶仕高 陈洁欣 匡海学[2] 王秋红 ZHOU Junfa;QIAO Tingting;ZHANG Zhihong;YE Shigao;CHEN Jiexin;KUANG Haixue;WANG Qiuhong(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006;Key Laboratory of Basic and Applied Research on Chinese Materia Medica,Ministry of Education,Heilongjiang University of Chinese Medicine,Key Laboratory of Chinese Medicinal Material Basis and Pharmacological Effect,Harbin 150040,China)

机构地区：[1]广东药科大学中药学院,广东广州510006 [2]黑龙江中医药大学省部共建教育部北药基础与应用研究重点实验室,黑龙江省中药及天然药物药效物质基础研究重点实验室,黑龙江哈尔滨150040

出　　处：《南京医科大学学报（自然科学版）》2024年第5期604-614,共11页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家重点研发计划项目(2018YFC1707100);国家自然科学基金(8177141630);国家“重大新药创制”科技重大专项(2018ZX09731-001)。

摘　　要：目的:探究胆汁酸单体化合物胆酸(cholic acid,CA)和去氧胆酸(deoxycholic acid,DCA)的抗惊厥作用机制。方法:将3周龄雄性SD大鼠随机分为对照组、模型组、丙戊酸钠组(sodium valproate/valproic acid,VPA,189 mg/kg)、CA组(60 mg/kg)和DCA组(60 mg/kg),每组9只,对照组及模型组为假给药,各给药组于造模前1 h预给药,连续给药16 d。采用(45.0±0.5)℃水浴建立惊厥大鼠模型,每隔1 d水浴1次,共计8次,观察记录模型组及各给药组大鼠惊厥发作时间、惊厥结束时间,对大鼠惊厥发作行为的严重程度进行评分。检测大鼠血清及海马组织中白介素1β(interleukin 1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor α,TNF-α)和IL-6含量及海马组织中谷氨酸(glutamic acid,Glu)和γ-氨基丁酸(γ-aminobutyric acid,GABA)含量,苏木精-伊红(hematoxylin-eosin,HE)染色观察海马神经元病理损伤。超高效液相色谱仪串联高分辨质谱技术对大鼠血清进行代谢组学分析。结果:与模型组相比,各给药组大鼠惊厥潜伏期均显著延长,惊厥持续时间显著缩短(P均<0.001);VPA组和DCA组惊厥发作等级显著降低(P <0.001,P <0.01),CA组差异无统计学意义。与对照组相比,模型组血清和海马组织中TNF-α,IL-6和IL-1β含量均显著升高(P <0.001),海马组织中Glu和GABA含量均显著升高(P <0.001);与模型组相比,DCA组和VPA组各项生化指标水平均显著降低(P <0.001),而与模型组相比,除血清中IL-1β和海马组织IL-6的水平外,CA组其他各项指标均显著降低(P <0.01)。海马组织HE染色结果显示,与对照组相比,模型组大鼠海马组织中锥体细胞胞体紧缩,体积变小,染色加深,嗜碱性增强,胞质胞核分界不清;与模型组相比,各给药组大鼠海马神经元细胞形态得到明显改善。其中,DCA组大鼠海马神经元细胞形态与VPA组相近。血清代谢组学分析结果经过数据处理、文献及数据库比对,共鉴定出312个差异化合物。通过主Objective:To investigate the anticonvulsant mechanisms of bile acid monomer compounds cholic acid(CA)and deoxycholic acid(DCA).Methods:Male SD rats were randomly divided into a control group,a model group,a sodium valproate or valproic acid(VPA),(189 mg/kg),a CA group(60 mg/kg),and a DCA group(60 mg/kg),with nine rats in each group.The rats in the control group and model group were given placebo,and the mice in each treatment group was pre-treated 1 h before modeling,and continuously treated for 16 d.A seizure rat model was established using a water bath at(45.0+0.5)C,with a bath given every other day for a total of eight times.The seizure onset time,seizure termination time,and the severity of seizure behavior of rats were observed and recorded.Meanwhile,the levels of interleukin 1β(IL-1β),interleukin 6(IL-6),and tumor necrosis factorα(TNF-α)in rat serum and hippocampal tissues,as well as the contents of glutamate(Clu)and-aminobutyric acid(GABA)in hippocampal tissues were detected.Hematoxylin-eosin(HE)staining was used to observe the pathological damage of hippocampal neurons.Metabolomic analysis of rat serum was performed using the ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS).Results:Compared with the model group,all treatment groups significantly prolonged the latency of seizures but significantly reduced the duration of seizures(P<0.001);both the VPA group and DCA group significantly reduced the severity of seizures(P<0.001,P<0.01),while there was no significant difference in the CA group.Compared with the control group,the contents of IL-1βTNF-α,and IL-6 in serum and hippocampal tissues of the model group were significantly increased(P<0.001),and the contents of Clu and GABA in hippocampus were also significantly increased(P<0.001).Compared with the model group,the effects produced by the DCA group and the VPA group were similar,both of which reduced the levels of various biochemical indicators(P<0.001),while the CA group significantly reduced all indicators except

关 键 词：胆酸 去氧胆酸 惊厥 代谢组学 代谢通路 

分 类 号：R971.6[医药卫生—药品]