Yutthana Chotritthirong

This study investigated the combined antioxidant effects of kaempferol (KAE) and N-acetylcysteine (NAC) against hydrogen peroxide (H₂O₂)-induced oxidative stress in SH-SY5Y cells. Combination effects were evaluated using combination index (CI) analysis and SynergyFinder. Cell viability, 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate (CM-H₂DCFDA) fluorescence, malondialdehyde (MDA), antioxidant enzyme activities, glutathione status, and protein expression of sirtuin 3 (SIRT3), forkhead box O3 (FOXO3a), and superoxide dismutase 2 (SOD2) were assessed. Molecular docking was performed to predict potential interactions of KAE and NAC with SIRT3. The strongest CI-defined synergy occurred at 16 μM KAE and 2 mM NAC (CI = 0.10), whereas SynergyFinder results varied across reference models. The combination (KNAC) enhanced cell viability, reduced CM-H₂DCFDA fluorescence and MDA levels, improved antioxidant enzyme activities and glutathione redox balance, and increased SIRT3, FOXO3a, and SOD2 protein levels compared with individual treatments. Molecular docking predicted a more favorable binding score for KAE than NAC against SIRT3. These findings suggest that KAE and NAC may produce complementary antioxidant and cytoprotective effects through potentially distinct mechanisms, with KAE potentially contributing to changes in proteins associated with mitochondrial antioxidant defense and NAC supporting glutathione-dependent cellular redox homeostasis. However, the causal involvement of the SIRT3/FOXO3a/SOD2 pathway and direct interactions with SIRT3 remain to be experimentally confirmed. These preliminary findings support further investigation of combining food-derived KAE with NAC as a glutathione precursor to enhance cellular antioxidant defenses.

This study investigated the synergistic potential of mulberry leaf extract and Trolox against hydrogen peroxide (H₂O₂)-induced oxidative stress in SH-SY5Y cells. Cells were treated with the combination for 24 h prior to exposure to 200 µM H₂O₂. Cell viability was assessed using the MTT assay, while oxidative status was evaluated through measurements of intracellular reactive oxygen species (ROS), malondialdehyde (MDA), and the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px). Sirtuin 1 (SIRT1) and apoptosis-related proteins, including p53, cyclic AMP response element-binding protein (CREB), Bcl-2-associated X protein (Bax), and B-cell lymphoma 2 (Bcl-2), were determined by Western blot analysis. The combination treatment markedly upregulated SIRT1 expression, which was associated with increased CREB and Bcl-2 expression alongside reduced p53 and Bax levels. Increased SIRT1 expression was also accompanied by significant reductions in ROS and MDA levels and restoration of antioxidant enzyme activities. Collectively, these effects contributed to attenuation of oxidative stress and apoptosis, resulting in improved cell viability. These findings may support the formulation as a promising functional food-based strategy against oxidative stress-induced neuronal damage. However, these results are based on an in vitro SH-SY5Y cell model and represent preliminary evidence, warranting further in vivo and clinical studies to confirm their translational potential.

Background/Objectives: Unpredictable chronic mild stress exposure is a primary driver of cognitive decline, largely mediated by hypothalamic–pituitary–adrenal (HPA) axis dysregulation and subsequent oxidative neurotoxicity. In traditional Thai medicine, the AYW-KK-04 formulation—a complex polyherbal remedy—has long been utilized as a “Ya Aayu-Wattana” to restore vitality and elemental balance, yet its neurobiological mechanisms remain poorly understood. This study aimed to evaluate the adaptogenic and neuroprotective potential of AYW-KK-04 against cognitive impairment. Methods: Unpredictable Chronic Mild Stress (UCMS)-induced cognitive impairment in a ICR mouse model. Total phenolic and flavonoid contents and antioxidant capacity (ABTS assay) of AYW-KK-04 were determined. Behavioral assessments using Y-maze test, novel object recognition test (NORT), and Morris Water Maze (MWM) test. BDNF, CREB, Nrf and Keap1 mRNA gene expression, SOD and CAT enzymatic activity and lipid peroxidation assay were investigated to clarify the mechanisms of action. Moreover, HPLC chromatography was studied to quantify the active compounds of the AYW-KK-04 formulation. Results: It demonstrated that oral administration of AYW-KK-04 significantly reversed UCMS-induced memory deficits. At the molecular level, AYW-KK-04 effectively upregulated BDNF and CREB mRNA expression in the frontal cortex and hippocampus, suggesting a restoration of synaptic plasticity. Simultaneously, the formulation activated the Nrf2/Keap1 signaling pathway, leading to enhanced SOD and CAT enzymatic activities and a marked reduction in MDA-mediated lipid peroxidation. HPLC analysis confirmed the presence and consistency of key bioactive constituents. Conclusions: These findings suggest that the adaptogenic properties of AYW-KK-04 arise from its dual capacity to reinforce neurotrophic support and bolster the endogenous antioxidant shield, providing a mechanistic support for the traditional use of AYW-KK-04 as an adaptogenic formulation and highlighting its potential as a multi-target intervention for stress-related cognitive dysfunction.

Although Oroxylum indicum is a widely consumed herb with many pharmaceutical qualities, no study has investigated its sensory properties, especially the bitterness. To thoroughly understand the bitter taste, we conducted machine learning (BitterX) screening of bitter taste receptors for beneficial flavonoids in the fruits, explored the interactions through molecular docking, and performed chemical quantification with electronic tongue analysis to determine the compounds contributing to the overall bitterness. The main flavonoids were found to bind to five bitter taste receptors with high predicted scores and low binding affinities, including TAS2R5, TAS2R7, TAS2R14, TAS2R39, and TAS2R43. Through multivariate data analysis, baicalein, baicalin, chrysin, oroxin A, oroxin B, and chrysin-7-O-glucuronide were highly correlated with the overall bitterness, ranging from R = 0.83 to 0.92. These compounds were present more in the seeds, making them more bitter than the coats. The principal component analysis biplot also indicated that saltiness could possibly mask the bitterness of O. indicum.

The resin extract of Garcinia nigrolineata (GNR-E), a tropical plant used in Southeast Asian traditional medicine, was evaluated for its antidepressant-like effects in a chronic mild stress (CMS) mouse model, with imipramine as a reference drug. GNR-E dose-dependently alleviated CMS-induced anhedonia (sucrose preference test) and behavioral despair (forced swimming and tail suspension tests). Neurochemical analyses revealed that GNR-E increased serotonin (5-HT) and norepinephrine (NE) levels, reduced expression of their transporters (SERT, NET) and receptors (5-HT1A, 1B, 2A, 2C, 7; α2A, 2C) in the frontal cortex and hippocampus, and normalized HPA-axis hyperactivity by lowering serum corticosterone and modulating glucocorticoid receptor (GR) and SGK-1 mRNA expression. In vitro, GNR-E inhibited monoamine oxidase (MAO)-A and -B (Ki = 2.33 and 1.55 µg/mL, respectively). Phytochemical analysis identified xanthones, particularly cowanin, as key constituents. These findings highlight GNR-E’s potential as a novel plant-based antidepressant, warranting further investigation into its active compounds and clinical applications.

Background Chronic stress exposure has been widely recognized as a significant contributor to numerous central nervous system (CNS) disorders, leading to debilitating behavioral changes such as anxiety, depression, and cognitive impairments. The prolonged activation of the hypothalamic–pituitary–adrenal (HPA) axis during chronic stress disrupts the neuroendocrine balance and has detrimental effects on neuronal function and survival. Nelumbo nucifera (N. nucifera) Gaertn., commonly known as the lotus flower, is a traditional medicinal plant consumed for its purported benefits on mental and physical well-being. Despite its traditional use, limited scientific evidence supports these claims. Methods The present study explores the effects of N. nucifera, commonly known as the lotus flower, on cognitive performance and stress resilience in a mouse model subjected to unpredictable chronic mild stress (UCMS). Results Daily treatment significantly improved cognitive performance, alleviated depressive-like behaviors, and normalized hypothalamic–pituitary–adrenal (HPA) axis activity, as indicated by a 60.97% reduction in serum corticosterone. At the molecular level, N. nucifera petals also downregulated serum- and glucocorticoid-inducible kinase 1 (SGK1) mRNA expression while upregulating brain-derived neurotrophic factor (BDNF) mRNA expression and cyclic-adenosine monophosphate (cAMP) responsive element-binding protein (CREB) mRNA expression in the hippocampus and frontal cortex. These normalizations are critical, as chronic stress dysregulates HPA axis function, exacerbating behavioral changes. Furthermore, a phytochemical analysis resulted in the isolation of five major compounds, kaempferol (1), trifolin (2), kaempferol-3-neohesperidoside (3), icariside D₂ (4), and β-sitosterol (5), each demonstrating significant monoamine oxidase (MAO) inhibitory activity. Conclusions These compelling findings suggest that N. nucifera petals not only alleviate stress-induced mood and cognitive deficits but also offer a promising avenue for modulating the HPA axis and promoting neuroprotection via essential neurotrophic factors and enzymatic pathways. We advocate for its potential as a complementary and alternative medicine for effective stress management. Future investigations should further explore its mechanisms of action and evaluate its clinical applicability in stress-related disorders.

Postmenopausal women have a higher probability of experiencing cognitive alterations compared to men, suggesting that the decline in female hormones may contribute to cognitive deterioration. Thailand traditionally uses Tri-Kaysorn-Mas (TKM), a blend of three medicinal herbs, as a tonic to stimulate appetite and relieve dyspepsia. Due to its antioxidant and anti-acetylcholinesterase activities, we investigated the effects of TKM (50 and 100 mg/kg/day, p.o., for 8 weeks) on cognitive deficits and their underlying causes in an ovariectomized (OVX) mouse model of menopause. OVX mice showed cognitive impairment in the Y-maze, novel object recognition task (NORT), and Morris water maze (MWM) behavioral tests, along with atrophic changes to the uterus, altered levels of serum 17β-estradiol, and down-regulated expression of estrogen receptors (ERα and ERβ). These behavioral effects were reversed by TKM. TKM decreased malondialdehyde (MDA) levels and mitigated oxidative stress in the brain by enhancing the activity of superoxide dismutase (SOD) and catalase (CAT) and by up-regulating the antioxidant-related gene Nrf2 while down-regulating Keap1. TKM also counteracted OVX-induced neurodegeneration by enhancing the expression of the neurogenesis-related genes BDNF and CREB. The results indicate that TKM extract alleviates oxidative brain damage and neurodegeneration while enhancing cognitive behavior in OVX mice, significantly improving cognitive deficiencies related to menopause/ovariectomy through multiple targets.

The Trikaysornmas formula (TKM) represents a prevalent Thai traditional remedy utilized extensively in Thailand. Its traditional uses include appetite enhancement, functions as a nourishing tonic, and exhibits adaptogenic properties. Comprising Aegle marmelos fruit, Nelumbo nucifera stamen, and Jatropha multifida bark, this formula embodies the synergy among these three herbs. The objective of this study was to optimize the extraction method, determine the active compounds in the TKM, and evaluate its antioxidant activity. The optimization of the extraction method for this formula was studied using an experimental design. Phytochemical components such as total phenolics, total flavonoids, total carotenoids, and total alkaloids were assessed utilizing a colorimetric method. Antioxidant activities were assessed through DPPH free radical scavenging, ABTS radical cation decolorization, oxygen radical absorbance capacity, ferric reducing antioxidant power, metal chelating activity, and lipid peroxidation assay. For the analysis of active constituents in the formula, gallic acid, kaempferol‐3‐o‐glucoside, imperatorin, vitexin, and scopoletin, a validated reversed‐phase column high‐performance liquid chromatography (HPLC) method was developed. The total active contents including phenolic, flavonoid, carotenoid, and alkaloid compounds were found in the formula. The developed HPLC method exhibited reliable results in all validation parameters. TKM demonstrated antioxidant activity in the models used in this research. The findings from this study can serve as valuable tools for standardization and quality control measures. Additionally, they can contribute to maximizing the possibilities inherent in this traditional Thai formulation.

Major depressive disorder (MDD) is one life-threatening disorder that is prevalent worldwide. The evident etiology of this disease is still poorly understood. Currently, herbal medicine is gaining more interest as an alternative antidepressant. Oroxylum indicum, which is used in traditional medicine and contains a potential antidepressive compound, baicalein, could have an antidepressive property. An in vitro monoamine oxidase-A (MAO-A) inhibitory assay was used to preliminarily screening for the antidepressant effect of O. indicum seed (OIS) extract. Mice were subjected to unpredictable chronic mild stress (UCMS) for 6 weeks, and the daily administration of OIS extract started from week 4. The mechanisms involved in the antidepressive activity were investigated. The OIS extract significantly alleviated anhedonia and despair behaviors in the UCMS-induced mouse model via two possible pathways: (i) it normalized the HPA axis function via the restoration of negative feedback (decreased FKBP5 and increased GR expressions) and the reduction in the glucocorticoid-related negative gene (SGK-1), and (ii) it improved neurogenesis via the escalation of BDNF and CREB expressions in the hippocampus and the frontal cortex. In addition, an HPLC analysis of the OIS extract showed the presence of baicalin, baicalein, and chrysin as major constituents. All of the results obtained from this study emphasize the potential of OIS extract containing baicalin and baicalein as an effective and novel alternative treatment for MDD.

Supercritical fluid extraction (SFE) is an innovative green technology for the extraction of phytochemicals from plants. Therefore, this study aimed to evaluate the application of SFE and to optimize the extraction conditions of the Thai herbal formula, Kleeb Bua Daeng (KBD). A Box–Behnken design (BBD) with response surface methodology (RMS) was used to determine the effect of the extraction time (30–90 min), temperature (30–60 °C), and pressure (200–300 bar) on response variables including the extraction yield, total phenolic content (TPC), total flavonoid content (TFC), total carotenoid content (TCC), and total anthocyanin content (TAC) of the KBD formula. The highest percentage extraction yield (3.81%) was achieved at 60 °C, 300 bar, and 60 min of the extraction time. The highest TPC (464.56 mg gallic acid equivalents/g extract), TFC (217.19 mg quercetin equivalents/g extract), and TCC (22.26 mg β-carotene equivalents/g extract) were all achieved at 60 °C, 250 bar, and 90 min of the extraction time. On the contrary, it was not possible to quantify the total anthocyanin content as anthocyanins were not extracted by this method. The results indicated that SFE-CO₂ is a suitable method of extraction for a green recovery of phytochemicals with low and moderate polarity from the KBD formula.