Network pharmacology, molecular docking, and experimental validation to explore the potential mechanism of Long Mu Qing Xin mixture for the treatment of attention deficit hyperactivity disorder

Abstract

Background: Long Mu Qing Xin Mixture (LMQXM) has shown potentially positive effects in alleviating attention deficit hyperactivity disorder (ADHD); however, the action mechanism is still not fully understood. This study aimed to predict the potential mechanism of LMQXM for ADHD using network pharmacology and molecular docking, which were then validated using animal experiments.Methods: Network pharmacology and molecular docking techniques were used to predict the core targets and potential pathways of LMQXMQ for ADHD, and KEGG pathway enrichment analysis revealed the potential significance of dopamine (DA) and cyclic adenosine monophosphate (cAMP) signaling pathways. To verify the hypothesis, we conducted an animal experiment. In the animal experiment, the young spontaneously hypertensive rats (SHRs) were randomly divided into the model group (SHR), the methylphenidate hydrochloride group (MPH, 4.22 mg/kg), and 3 LMQXM groups (low-dose (LD) group, 5.28 ml/kg; medium-dose (MD) group, 10.56 ml/kg; and high-dose (HD) group, 21.12 ml/kg), and administered by gavage for 4 weeks; the WKY rats were set as the control group. The open field test and Morris water maze test were used to evaluate the behavioral performance of rats, high performance liquid chromatography mass spectrometry (LC-MS) was used to analyze DA levels in the prefrontal cortex (PFC) and striatum of rats, ELISA was used to detect cAMP concentrations in the PFC and striatum, and immunohistochemistry and qPCR were used to analyze positive cell expression and mRNA expression for indicators related to DA and cAMP pathways.Results: The results showed that beta-sitosterol, stigmasterol, rhynchophylline, baicalein, and formononetin might be key components of LMQXM for ADHD and that these components bind well to the core targets, DA receptors (DRD1 and DRD2). Furthermore, LMQXM might act through the DA and cAMP signaling pathways. In the animal experiment, we found that MPH and LMQXM-MD controlled hyperactivity and improved learning and memory in SHRs, while LMQXM-HD only controlled hyperactivity in SHRs; meanwhile, MPH and LMQXM-MD upregulated DA and cAMP levels, mean optical density (MOD) of cAMP, and MOD and mRNA expression of DRD1 and PKA in the prefrontal cortex (PFC) and striatum of SHRs, while LMQXM-LD and LMQXM-HD upregulated DA and cAMP levels in the striatum, MOD of cAMP in the PFC, and mRNA expression of PKA in the PFC. However, we did not find a significant regulatory effect of LMQXM on DRD2.Conclusion: To sum up, this study demonstrated that LMQXM may increase DA levels mainly by activating the cAMP/PKA signaling pathway through DRD1, thereby controlling the behavioral disorders of SHRs, which is most effective at moderate doses, and this may be a key mechanism for LMQXM in the treatment of ADHD.

Exploring the HIF-1α signalling pathway and the mechanism of YiQiHuoXue decoction against Precancerous Lesions of Gastric Cancer based on Network Pharmacology and Molecular Docking

“…Results showed that the above 6 active compounds were able to spontaneously bind to the 3 targets naturally, ligands without 3D structure were excluded (Diop, methylicosa-11,14-dienoate, Mandenol). The conformation of the compound binding to the target is more stable the lower the binding energy 12 .…”

Long Mu Qing Xin mixture improves behavioral performance in spontaneously hypertensive rats (SHR/NCrl) by upregulating catecholamine neurotransmitters in prefrontal cortex and striatum via DRD1/cAMP/PKA-CREB signaling pathway

“…The clinically effective dose of LMQXM for children was 75 mL/d, which was concentrated to three-fifths of the original volume, i.e., 45 mL/d, prior to the experiment, taking into account the limitations of the animals’ gavage volume. The dose administered to the animals was based on the body surface area of a 9-year-old (body mass 26 kg) child (0.991 m 2 , Stevenson’s formula = 0.0061×height (cm)+0.0128×body weight (kg)-0.1529) and 4-week-old (body mass 60 g) SHR pups body surface area (0.013947 m 2 , Meeh-Rubner formula = 9.1 × body weight (g)⅔/10,000) were converted in parallel ( Song et al, 2021 ; Li et al, 2023 ). The clinically equivalent dose of SHR/NCrl rats was calculated to be 10.56 mL/kg/d.…”

“…Network pharmacology analyses have proposed the cAMP signalling pathway as a probable key avenue through which LMQXM exerts its therapeutic action in ADHD. Furthermore, pharmacological inquiries have corroborated LMQXM’s ability to ameliorate the fundamental symptoms of ADHD by elevating dopamine (DA) levels within the prefrontal cortex (PFC) and striatum of spontaneously hypertensive rats (SHRs) ( Li et al, 2023 ). However, the specific mechanism underlying LMQXM’s influence on catecholamine neurotransmitters necessitates further elucidation.…”