Table 1_Mechanisms of Changma Xifeng tablet in alleviating Tourette syndrome via modulation of neurotransmitters, inflammatory responses, and metabolic pathways.docx

IntroductionTourette syndrome (TS) is a common pediatric neurodevelopmental disorder. Changma Xifeng tablet (CMXF), a traditional Chinese medicinal formulation, is widely used clinically for TS but its mechanisms remain unclear. This study aimed to evaluate CMXF’s efficacy and explore its action mechanisms via a TS murine model.
MethodsThe chemical constituents of CMXF were analyzed and identified by UPLC-Q-TOF-MS/MS. Sixty male BALB/c mice were divided into 6 groups (CON, MOD, HAL, CMXF-L/M/H). TS models were induced by IDPN injection. Behavioral assessments, ELISA (neurotransmitters/inflammatory cytokines), HE staining (striatal pathology), RT-qPCR/WB (DRD1/DRD2/COMT/MAO-B), and UHPLC-Q-TOF LC-MS (serum metabolomics) were performed.
ResultsA total of twenty-one components, including gallic acid, gastrodin, catechin, sibiricose A3, paeoniflorin, parishin B, and tenuifoliside B, were identified in CMXF. From week 4 to week 8, the HAL group and CMXF-H group showed significantly improved behavioral performance compared with the MOD group (F > 1, P < 0.001), presenting a dose-dependent trend. ELISA results revealed that CMXF-H significantly increased the levels of 5-HT in serum and striatum (F > 1, P < 0.001), while decreasing the levels of DA, HVA, NE, IL-1β, and IL-6 in serum (F > 1, P < 0.001). RT-qPCR and WB results indicated that CMXF regulated the mRNA and protein, inhibiting DRD1/DRD2 expressions and inducing COMT/MAO-B expressions. Serum metabolomics analysis identified 398 differentially expressed metabolites, which were mainly involved in lipid metabolism and organic acid metabolism pathways.
DiscussionCMXF exerts anti-TS effects by regulating neurotransmitters, inflammatory cytokines, DA signaling, and serum metabolites, providing novel insights into its mechanisms and potential targeted therapy for TS.

引言：抽动秽语综合征（Tourette syndrome, TS）是一种常见的儿童神经发育障碍。长麻熄风片（Changma Xifeng tablet, CMXF）作为一种中药复方制剂，在临床中被广泛用于抽动秽语综合征的治疗，但其具体作用机制尚未明确。本研究旨在通过抽动秽语综合征小鼠模型，评价长麻熄风片的疗效并探究其作用机制。
方法：采用超高效液相色谱-串联四极杆飞行时间质谱（Ultra Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry, UPLC-Q-TOF-MS/MS）对长麻熄风片的化学成分进行分析与鉴定。将60只雄性BALB/c小鼠随机分为6组，分别为对照组（CON）、模型组（MOD）、氟哌啶醇组（HAL）、长麻熄风片低剂量组（CMXF-L）、中剂量组（CMXF-M）与高剂量组（CMXF-H）。通过注射3-碘丙基腈（3-Iodopropionitrile, IDPN）构建抽动秽语综合征小鼠模型。随后开展多维度实验检测：包括行为学评估、酶联免疫吸附实验（Enzyme-Linked Immunosorbent Assay, ELISA，检测神经递质与炎性细胞因子）、苏木精-伊红染色（Hematoxylin-Eosin staining, HE staining，观察纹状体病理形态）、实时荧光定量聚合酶链反应（Real-Time Quantitative Polymerase Chain Reaction, RT-qPCR）与蛋白质印迹（Western Blot, WB，检测多巴胺D1受体、多巴胺D2受体、儿茶酚-O-甲基转移酶以及单胺氧化酶B的表达水平）以及超高效液相色谱-串联四极杆飞行时间液相质谱（Ultra High Performance Liquid Chromatography-Quadrupole Time-of-Flight Liquid Chromatography-Mass Spectrometry, UHPLC-Q-TOF LC-MS，开展血清代谢组学分析）。
结果：本研究共从长麻熄风片中鉴定出21种化学成分，包括没食子酸（Gallic acid）、天麻素（Gastrodin）、儿茶素（Catechin）、sibiricose A3、芍药苷（Paeoniflorin）、parishin B以及tenuifoliside B。在造模后第4周至第8周，氟哌啶醇组与长麻熄风片高剂量组的行为学表现均显著优于模型组（F>1，P<0.001），且呈现出剂量依赖性趋势。酶联免疫吸附实验结果显示，长麻熄风片高剂量组可显著升高血清与纹状体中5-羟色胺（5-Hydroxytryptamine, 5-HT）的水平（F>1，P<0.001），同时降低血清中多巴胺（Dopamine, DA）、高香草酸（Homovanillic acid, HVA）、去甲肾上腺素（Norepinephrine, NE）、白细胞介素-1β（Interleukin-1β, IL-1β）以及白细胞介素-6（Interleukin-6, IL-6）的水平（F>1，P<0.001）。实时荧光定量聚合酶链反应与蛋白质印迹实验结果表明，长麻熄风片可调控相关基因的mRNA与蛋白表达：抑制多巴胺D1受体（Dopamine Receptor D1, DRD1）与多巴胺D2受体（Dopamine Receptor D2, DRD2）的表达，同时促进儿茶酚-O-甲基转移酶（Catechol-O-Methyltransferase, COMT）与单胺氧化酶B（Monoamine Oxidase B, MAO-B）的表达。血清代谢组学分析共鉴定出398个差异表达代谢物，这些代谢物主要参与脂质代谢与有机酸代谢通路。
讨论：长麻熄风片可通过调控神经递质、炎性细胞因子、多巴胺信号通路以及血清代谢物发挥抗抽动秽语综合征的作用，本研究为阐明其作用机制以及开发抽动秽语综合征的潜在靶向治疗手段提供了全新的研究视角。