龙牡宁心方治疗注意缺陷多动障碍的疗效与机制探讨

Exploration of the efficacy and mechanism of Longmu Ningxin Formula for attention deficit hyperactivity disorder

摘要

目的: 探讨龙牡宁心方治疗儿童注意缺陷多动障碍(attention deficit hyperactivity disorder, ADHD)的临床疗效,并结合动物实验与蛋白质组学探究其潜在作用机制。方法: 将80例ADHD患儿随机分为西药组和中药组各40例,西药组给予盐酸哌甲酯缓释片,中药组给予龙牡宁心方,治疗持续12周,比较两组患儿治疗前后SNAP-IV评定量表(父母版)、Weiss功能缺陷量表(父母版)、Conners儿童行为量表(父母版)评分与中医证候积分,并进行安全性评价。采用ADHD的经典模型自发性高血压大鼠(spontaneously hypertensive rat,SHR)进行动物实验,将18只4周龄雄性SHR随机分为模型组、中药组、西药组,6只Wistar-Kyoto大鼠作为对照组;中药组给予38.00 g·kg^－1·d^－1龙牡宁心方,西药组给予4.22 mg·kg^－1·d^－1盐酸哌甲酯,对照组与模型组灌胃等量生理盐水,连续4周,行为学测试评估大鼠行为表现,实验结束后取前额叶组织进行定量蛋白质组学分析,并进行Western Blot分析验证差异表达蛋白。结果: 治疗后,中药组的SNAP-IV评定量表、Weiss功能缺陷量表、Conners儿童行为量表评分、中医主症与次症积分均较治疗前显著下降(P均＜0.001);中药组的Weiss功能缺陷量表、Conners儿童行为量表评分、中医证候积分改善程度均大于西药组(P均＜0.05);所有患儿在治疗期间均未出现与研究药物相关的严重不良反应。龙牡宁心方显著改善了SHR的多动冲动与空间学习记忆缺陷(P均＜0.05)。模型组与中药组之间的前额叶蛋白质组学共鉴定到273种差异表达蛋白质,主要涉及NOD样受体信号通路、FoxO信号通路、肠道免疫网络与IgA合成等7条途径。Western Blot结果显示龙牡宁心方逆转了SHR前额叶中上调的Ripk2、Tab1与IL-6。结论: 龙牡宁心方治疗儿童ADHD疗效较好,且安全性较高,其作用机制可能为调控前额叶Ripk2、Tab1、IL-6表达抑制NOD样受体信号通路介导的神经炎症。

Abstract

Objective: To investigate the clinical efficacy of Longmu Ningxin Formula (LMNXF) in treating children with attention deficit hyperactivity disorder (ADHD) and to explore its potential mechanism of action through animal experiments and proteomics. Methods: A total of 80 children with ADHD were randomly divided into a western medicine group (n=40) and a Chinese medicine group (n=40). The western medicine group was treated with methylphenidate hydrochloride extended-release tablets (MPH), while the Chinese medicine group received LMNXF. The treatment lasted for 12 weeks. The scores of the Swanson, Nolan, and Pelham-Ⅳ Rating Scale (SNAP-Ⅳ, parent version), the Weiss Functional Impairment Rating Scale (WFIRS, parent version), the Conners' Parent Rating Scale (CPRS), and the Chinese medicine syndrome scores were compared between the two groups before and after treatment. Safety was also evaluated. For the animal experiment, the spontaneously hypertensive rat (SHR), a classic ADHD model, was used. A total of 18 4-week-old male SHRs were randomly divided into a model group, a Chinese medicine group, and a western medicine group. A total of 6 Wistar-Kyoto rats served as the control group. The Chinese medicine group received LMNXF at 38.00 g·kg^－1·d^－1, the western medicine group received MPH at 4.22 mg·kg^－1·d^－1, and the control and model groups were administered an equal volume of saline by gavage. Treatment lasted for 4 weeks. Behavioral tests were conducted to assess the rats' performance. After the experiment, prefrontal cortex tissues were collected for quantitative proteomic analysis, and Western Blot analysis was performed to validate differentially expressed proteins. Results: After treatment, the scores of the SNAP-Ⅳ, WFIRS, CPRS, and the primary and secondary Chinese medicine syndrome scores in the Chinese medicine group were significantly decreased compared to those before treatment (all P<0.001). The improvements in the WFIRS and CPRS scores and the Chinese medicine syndrome scores in the Chinese medicine group were greater than those in the western medicine group (all P<0.05). No serious adverse reactions related to the study drugs were observed in any child during the treatment period. LMNXF significantly ameliorated hyperactivity/impulsivity and spatial learning/memory deficits in SHRs (all P<0.05). Proteomic analysis of the prefrontal cortex identified 273 differentially expressed proteins between the model group and the Chinese medicine group. These proteins were primarily involved in seven pathways, including the NOD-like receptor signaling pathway, FoxO signaling pathway, and intestinal immune network for IgA production. Western Blot results showed that LMNXF reversed the upregulation of Ripk2, Tab1, and IL-6 in the prefrontal cortex of SHRs. Conclusion: LMNXF demonstrates favorable efficacy and a high safety profile in treating children with ADHD. Its mechanism of action may involve modulating Ripk2, Tab1, and IL-6 expression in the prefrontal cortex, thereby inhibiting NOD-like receptor signaling pathway-mediated neuroinflammation.

关键词

龙牡宁心方 / 注意缺陷多动障碍 / 临床研究 / 定量蛋白质组学

Key words

Longmu Ningxin Formula / attention deficit hyperactivity disorder / clinical research / quantitative proteomics

[1] GALLO EF, POSNER J. Moving towards causality in attention-deficit hyperactivity disorder: overview of neural and genetic mechanisms[J]. Lancet Psychiatry, 2016, 3(6): 555-567.
[2] KELLY BD. Attention-deficit hyperactivity disorder: a clinical review of the concept, diagnosis and management[J]. Ir J Psychol Med, 2018, 35(3): 157-161.
[3] NAGUY A. Psychopharmacotherapy of attention deficit-hyperactivity disorder in children with comorbid conditions[J]. Pediatr Neurol, 2018, 82: 7-12.
[4] NIMMO-SMITH V, MERWOOD A, HANK D, et al. Non-pharmacological interventions for adult ADHD: a systematic review[J]. Psychol Med, 2020, 50(4): 529-541.
[5] POSNER J, POLANCZYK GV, SONUGA-BARKE E. Attention-deficit hyperactivity disorder[J]. Lancet, 2020, 395(10222): 450-462.
[6] MECHLER K, BANASCHEWSKI T, HOHMANN S, et al. Evidence-based pharmacological treatment options for ADHD in children and adolescents[J]. Pharmacol Ther, 2022, 230: 107940.
[7] 鲍梦依, 赵家靖, 严晨毓, 等. 儿童ADHD数字药物的神经机制、应用进展与临床挑战[J]. 中国现代应用药学, 2025, 42(22): 3945-3953.
[8] 王新月, 王昕, 杨健. 经颅直流电刺激应用于儿童青少年注意缺陷多动障碍治疗研究进展[J]. 中国实用儿科杂志, 2025, 40(10): 853-859.
[9] 美国精神医学学会. 精神障碍诊断与统计手册[M]. 第5版. 伦敦: 美国精神医学学会, 2013: 59-65.
[10] 中华中医药学会. 中医儿科临床诊疗指南[M]. 北京: 中国中医药出版社, 2020.
[11] LI YQ, YUAN HX, SONG YC, et al. An Shen Ding Zhi Ling ameliorates the symptoms of attention deficit hyperactivity disorder via modulating brain-derived neurotrophic factor-related signaling pathways[J]. Evid Based Complement Alternat Med, 2022, 2022: 5471586.
[12] HORN JL, JANICKI PK, FRANKS JJ. Diminished brain synaptic plasma membrane Ca²⁺-atpase activity in spontaneously hypertensive rats: Association with reduced anesthetic requirements[J]. Life Sci, 1995, 56(22): PL427-PL432.
[13] 卢凤凤, 卢学春, 周培岚, 等. 基于生物信息学的右美托咪定药理学机制研究[J]. 军事医学, 2021, 45(2): 104-109.
[14] 袁海霞, 倪新强, 韩新民, 等. 熟地黄对ADHD模型大鼠前额叶皮质线粒体的保护作用[J]. 中成药, 2020, 42(8): 2025-2031.
[15] 赵子瑜, 刘启玲, 姚金余, 等. 低氧导致小鼠注意缺陷多动障碍样行为及其突触损伤机制[J]. 空军军医大学学报, 2023, 44(3): 229-234.
[16] 章嘉琦, 孙茹欣, 杨宇婷, 等. 基于NF-κB-NLRP3通路探讨补肾开窍方对注意缺陷多动障碍大鼠神经炎症的干预机制[J]. 广东医学, 2025, 46(6): 801-809.
[17] 李雪军, 王淑敏, 张奕星, 等. 血液炎症标志物水平与儿童注意缺陷多动障碍关系的Meta分析[J]. 医学研究杂志, 2023, 52(11): 74-83.
[18] 曾美芳. NOD2-RIPK2对化学物诱导小鼠肺癌发生和培养肺癌细胞增殖、迁移的影响[D]. 福州: 福建医科大学, 2022.
[19] BAO SY, SUN QX, YAO CL. The interaction of TAK1 and TAB1 enhances LPS-induced cytokine release via modulating NF-κB activation (Larimichthys crocea)[J]. Fish Shellfish Immunol, 2018, 74: 450-458.
[20] XU Y, LIN F, LIAO GL, et al. Ripks and neuroinflammation[J]. Mol Neurobiol, 2024, 61(9): 6771-6787.
[21] 黎鑫. NOD2-RIPK2通路在MPTP诱导的小鼠帕金森病模型的作用[D]. 福州: 福建医科大学, 2022.
[22] XU YR, LEI CQ. TAK1-TABs complex: a central signalosome in inflammatory responses[J]. Front Immunol, 2021, 11: 608976.
[23] DU RH, ZHOU Y, XIA ML, et al. α-Synuclein disrupts the anti-inflammatory role of Drd2 via interfering β-arrestin2-TAB1 interaction in astrocytes[J]. J Neuroinflammation, 2018, 15(1): 258.

基金资助

上海市卫生健康委员会科研项目计划(202240158);上海中医药大学附属龙华医院科技创新基础研究项目(CX202514);上海市科学技术委员会“科技创新行动计划”医学创新研究专项项目(23Y21920100);2025年度上海市自然科学基金项目(25ZR1402487);上海中医药大学科技发展项目(24KFL054)