Page 17 - 南京医科大学自然版

第44卷第5期 周俊发，乔婷婷，张志宏，等. 胆酸和去氧胆酸抗惊厥作用的血清代谢组学研究［J］.
2024年5月 南京医科大学学报（自然科学版），2024，44（5）：604-614 ·605 ·

recorded. Meanwhile，the levels of interleukin 1β（IL⁃1β），interleukin 6（IL⁃6），and tumor necrosis factor α（TNF⁃α）in rat serum and
hippocampal tissues，as well as the contents of glutamate（Glu）and γ ⁃ 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⁃1β TNF⁃α，
and IL⁃6 in serum and hippocampal tissues of the model group were significantly increased（P < 0.001），and the contents of Glu 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 the TNF⁃α level in serum and the IL⁃6 level in the hippocampus（P < 0.01）. HE
staining results of hippocampal tissues showed that compared with the control group，the pyramidal cells in the hippocampus of rats in
the model group were contracted，with a smaller volume，a darker staining，an enhanced alkalinity，and the unclear cytoplasmic nuclear
boundaries；compared with the model group，the morphology of hippocampal neurons in each treatment group was significantly
improved. Among them，the morphology of hippocampal neurons in the DCA group was similar to that in the VPA group. A total of 312
differential compounds were identified in serum metabolomics analysis. Through principal component analysis（PCA）and orthogonal
partial least squares⁃discriminant analysis（OPLS⁃DA）analysis，nine differential compounds were selected. The results of metabolic
pathway enrichment showed that the anticonvulsant effects of CA and DCA were mainly involved the citric acid cycle，amino acid
metabolism，and butyric acid metabolism pathways. Conclusion：CA and DCA have certain improvement effects on behavioral and
biochemical indicators of rats with febrile seizures，and their mechanisms of action may be correlated with energy metabolism，amino
acid metabolism，and short⁃chain fatty acid metabolism during seizures.
［Key words］ cholic acid；deoxycholic acid；seizure；metabolomics；metabolic pathways
［J Nanjing Med Univ，2024，44（05）：604⁃614］

热性惊厥（febrile seizure，FS）多发于儿童，是一 质发挥镇静抗惊厥作用暂不明确。动物胆汁相关
［1］
种体温突然升高并伴随癫痫发作的神经性疾病 ，复 的药物中多以胆汁酸类成分为主，胆酸（cholic acid，
发风险较高。研究表明，第 1 次惊厥发作时年龄不 CA）和去氧胆酸（deoxycholic acid，DCA）含量丰富，
满12个月的婴幼儿，在随后1年内有高达50%的概 是其主要的胆汁酸类成分，同时两者也是机体产生
［2］
率再次出现惊厥发作 ，而反复持续发作的惊厥会 的内源性次级胆汁酸代谢成分，因此推测 CA 和
损伤海马功能，影响记忆和学习，甚至会增加成年 DCA 可能是动物胆汁类传统药物发挥抗惊厥作用
后癫痫发作和其他神经性疾病发作的风险 。目前 的活性物质之一，而 CA 和 DCA 在结构上仅 7α位相
［3］
FS的病理生理学机制尚不确切，现代研究观点多认 差一个羟基，因此探究两者在惊厥治疗方面可能存
为该疾病与遗传、神经炎症和免疫等因素有关。 在的活性及差异性是有必要的。目前对于胆酸类
地西泮、苯巴比妥、丙戊酸钠（sodium valproate/ 化合物抗惊厥作用的研究甚少，其抗惊厥药效不清
valproic acid，VPA）等药物常用于临床的抗惊厥治 楚，作用机制不明。因此本研究通过胆汁酸单体化
［4］
疗 ，但对于婴幼儿来说，该类药物的潜在不良反 合物CA和DCA预给药惊厥大鼠，探索CA和DCA对
应，如短暂性共济失调、呼吸抑制、心动过缓以及低 反复性热水浴刺激导致的惊厥大鼠的药理作用，并
血压等导致的危害甚至远超疾病本身，因此并不适 利用血清代谢组学方法探究CA和DCA的抗惊厥作
合婴幼儿惊厥的预防和长期使用 。中医重视整体 用机制，旨在为开发新的抗惊厥药物提供研究思路
［5］
观念，讲究辨证治疗，在疾病防治和新药开发中具 和理论依据。
有独特优势，其中动物胆汁和动物胆汁类药物应用
1 材料和方法
于惊厥治疗在我国拥有久远历史。动物胆汁包括猪
胆粉、牛胆粉、蛇胆粉和熊胆粉等，胆汁类药物如胆 1.1 材料
南星、化风丹、牛黄等 ［6-7］ ，它们均被证明具有镇静、 1.1.1 仪器
抗炎、抗惊厥等多种治疗作用，但其中何种活性物 ME104/02 型电子分析天平（梅特勒⁃托利多仪