Background/Objectives: Osteoporosis is a progressive systemic skeletal disease, with male osteoporosis emerging as a critical global concern due to high morbidity and mortality from fractures. This study investigated the anti-osteoporotic potential of CGAC-a herbal mixture of Cervus elaphus Linnaeus, Glycine max (L.) Merr., Angelica gigas Nakai, and Cnidium officinale Makino-and its underlying mechanisms in an orchiectomized (ORX) mouse model. Methods: C57BL/6J mice underwent ORX for 8 weeks, followed by CGAC administration (250 and 500 mg/kg) for an additional 8 weeks. Molecular mechanisms were further validated using MG63 osteoblastic and RAW 264.7 osteoclast assays. Results: ORX induced severe osteoporotic phenotypes, including significant reductions in bone mineral density (BMD) and trabecular microarchitecture. Notably, at the time point examined, ORX was associated with a suppressed bone remodeling state, reflected by reductions in both TRAP-positive osteoclasts and ALP-positive osteoblasts, together with lower serum BALP, CTX-1, and Gla/Glu-OC ratio. Conversely, CGAC normalized this stagnant state and restored physiological remodeling. This was accompanied by reduced marrow fat accumulation through the AMPK signaling axis, which upregulated Runx2 and downregulated PPAR-γ. In vitro results confirmed that CGAC promoted osteoblast differentiation and mineralization while suppressing RANKL-induced osteoclastogenesis. These actions suggest that CGAC may be involved in regulating Wnt/β-catenin signaling. Conclusions: Overall, CGAC is a promising therapeutic candidate for male osteoporosis, offering pharmacological benefits particularly relevant to aging populations.

BackgroundThe medicinal plant Cnidium officinale (CO) is widespread in Northeast Asia and vulnerable to heat stress. The naturally occurring composition of pharmacological ingredients of CO results in overall physiological consequences; therefore, it is crucial to have a comprehensive understanding of metabolic response to ambient heat in terms of acclimation to estimate how much CO is exposed to threatening environmental conditions.ResultsTranscriptome analysis is critical for understanding the consequences of long-term physiological adaptation of CO to abiotic stress. However, transcriptome analysis on this species, particularly under prolonged stress conditions, has remained limited. We employed a temperature gradient tunnel (TGT) to subject CO to high-temperature exposure for four months, enabling us to observe the cumulative effects of heat and assess its acclimation mechanisms.In the absence of genome sequencing data, we performed de novo transcriptome assembly and compared DEGs from temperature treatment plots of a TGT and a growth chamber (GC).Since interpreting transcriptomic data can be complex, we employed a sequential analytical approach, including DEG clustering, GO enrichment, KEGG pathway mapping, miRNA-target gene analysis, and multiple rounds of RNA sequencing validation. DEGs were classified into two categories: genes exhibiting significant fold changes and genes showing significant count changes rather than fold changes. Then, we analyzed the functional roles of DEGs to determine which pathways respond to ambient and stressful high temperatures and validated the findings through cross-comparison with GC. Additionally, we conducted miRNA analysis to investigate post-transcriptional regulation under high temperatures. CO grown under higher ambient temperatures exhibited slight upregulation of pathways related to protein stability and turnover, ABA biosynthesis, and energy production, such as photosynthesis and oxidative phosphorylation. However, under extreme heat stress, most metabolic pathways were downregulated except for those involved in transcription, translation, oxidative phosphorylation and the biosynthesis of cutin, suberin, and wax.ConclusionThis study demonstrated that proper clustering of genes based on expression levels and fold changes in two different experimental conditions, along with pathway mapping, may provide a comprehensive understanding of CO’s response to heat stress. These insights could contribute to future research on heat tolerance and crop improvement.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12864-025-12051-5.

Abstract In South Korea, the traditional use of Korean medicine is recognized by the government, with efforts being made to expand its inclusion in health insurance. However, unlike pharmaceutical drugs, traditional medicines are used in clinical practice without proper non-clinical and clinical trial validation, raising criticism and concerns. Mangeum-tang is commonly used for treating sequelae from stroke and neuralgia. The rhizome of Cnidium officinale MAKINO, included in Mangeum-tang, is known to reduce embryonic growth and development in rats and induce mild limb malformations, raising safety concerns. The causes of malformations are diverse, often linked to genotoxicity, prompting toxicity investigations. In vitro, Mangeum-tang did not induce genetic mutations or chromosomal aberration, and in vivo, it did not cause micronucleus formation or acute toxicity in hematopoietic cells. Therefore, Mangeum-tang does not appear to induce acute toxicity or genotoxicity. However, the studies conducted were short-term, and toxicity from repeated administration has not been confirmed. To ensure the safety of Mangeum-tang, further studies, including repeated-dose toxicity tests, carcinogenicity tests, reproductive toxicity tests, and additional research on the genotoxicity of Cnidium officinale MAKINO, are necessary.

Menopause is the natural period of aging in women induced by ovary deterioration, resulting in estrogen deficiency. We evaluated the antioxidative and anti-inflammatory properties of Cnidium officinale, Pueraria lobata Ohwi, and Leonurus japonicus (CPL) extracts on vascular endothelial dysfunction. After treatment, CPL extracts decreased serum lipid profiles, serum vasoactive substances, tail temperatures, and cardiovascular risk indices. In ovariectomized rats, vasodilation significantly increased, with an increase in endothelial nitric oxide synthase (eNOS) in the CPL200 and CPL500 groups compared with the OVX group (p < 0.05). The extracts also significantly reduced vascular cell adhesion protein 1 (VCAM-1) in the CPL50, CPL100, and CPL200 groups compared with the OVX group (p < 0.05, p < 0.01, and p < 0.001, respectively). Intercellular adhesion molecule 1 (ICAM-1) was also reduced in the CPL100 and CPL200 groups compared with the OVX group (p < 0.001 and p < 0.0001, respectively); this was achieved through the downregulation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and inducible nitric oxide (iNOS), which resulted in the synthesis of nuclear factor erythroid 2-related factor 2 (NRF2) and eNOS in HUVECs. Our results show that CPL extracts could provide cardioprotective effects against vascular endothelium dysfunction by decreasing inflammation and upregulating vasodilation, ascertained by evaluating the antioxidant systems of ovariectomized rats. Further studies are needed to explore the long-term cardioprotective effects.

ABSTRACT To reveal the chemical composition of Cnidium officinale Makino (COM), improve the quality control standard, and explore the related toxicity, solvent extraction, SiO 2 gel column chromatography, and nuclear magnetic resonance (NMR, 1 H&amp; 13 C) technology were used to extract, isolate, and identify the structures of the secondary metabolites from COM. A total of 10 batches of COM from different origins were studied by HPLC; the data from HPLC were analyzed by chemometry. At the same time, using zebrafish as an experimental animal model, the effects of different polar chemical parts of COM on zebrafish embryo development were explored. The five compounds were identified in the experiment, namely senkyunolide A, senkyunolide H, ligustrolide I, ferulic acid, and chlorogenic acid. The fingerprint of COM was established, and 19 common peaks were identified, seven of which were characteristic peaks. Toxicity studies of zebrafish embryo development showed that the five fractions ( n ‐hexane, chloroform, ethyl acetate, n ‐butanol, and water fractions) did not show significant toxicity when the dose concentration was between 0.5 and 1.5 mg/mL, but the toxicity was significant when the dose was up to 2.5 mg/mL. Zebrafish embryos showed different degrees of spinal curvature, pericardial edema, delayed yolk sac absorption, and developmental retardation. This research has preliminarily elucidated the chemical foundation of COM. It has successfully established the fingerprint‐based quality control standard for COM and put forward the safe application ranges of extracts from diverse chemical fractions. These achievements have furnished an experimental foundation for subsequent in‐depth investigations.

This study aims to construct essential information on the pests attacking Cnidium officinale Makino, which is one of the most important medicinal plants in Korea and neighboring countries. Based on the current survey, a total of 12 species were identified, including three above-ground pests attacking flowers, leaves, and stems, as well as ten soil pests attacking roots. In the vertical distribution of damaged roots, the dominant species is bulb mite (Rhizoglyphus robini) followed by onion maggot (Delia antiqua). Based on this study and the previous literature, the total number of species of pests reported to attack C. officinale is 36, including 3 on flowers, 16 on leaves, 6 on stems, and 11 on roots. We also investigated and compiled a list of natural enemies based on all available information and the current study, totaling 14 species. Parasitus sp., Macrocheles glaber, and Smicroplectrus sp. were identified as candidate natural enemies of root pests.

To explore the origin and evolution of Ligusticum Chuanxiong, we conducted a component analysis of Ligusticum Chuanxiong and its medicinal relatives. This study encompassed seven species from various origins, including Chuanxiong (Ligusticum chuanxiong Hort.), Gansu Chuanxiong (Ligusticum chuanxiong cv. Gansu), Yunnan Chuanxiong (Ligusticum chuanxiong cv. Yunnan), Japanese Chuanxiong (Cnidium officinale Makino), Fuxiong (Ligusticum sinense 'Fuxiong'), Gaoben (Ligusticum sinense), and Liaogaoben (Ligusticum jeholense), comprising 27 distinct materials. We employed headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) to identify various odor profiles from these species using electronic nose technology (E-nose). The method effectively identified volatile constituents in the leaves of these seven species. Results indicated that odor differences between L. chuanxiong and its medicinal relatives were predominantly observed in sensors W1W and W1S. Linear discriminant factor analysis (LDA) successfully distinguished five of the relatives; however, L. chuanxiong and L. sinense exhibited high odor similarity, limiting complete differentiation in some samples. HS-SPME-GC-MS identified a total of 118 volatile constituents, with eight differential volatiles identified: trans-Neocnidilide, β-Caryophyllene, β-Selinene, 5-Pentylcyclohexa-1,3-diene, (E)-Ligustilide, Butylphthalide, Neophytadiene, and Senkyunolide. Hierarchical cluster analysis (HCA) grouped L. chuanxiong, L. sinense, L. jeholense, and L. chuanxiong cv. Gansu together, highlighting the close relationship between L. chuanxiong and L. sinense. Joint analysis revealed a significant positive correlation between sensor W1W and the differential volatile component β-Caryophyllene, suggesting its potential for distinguishing closely related species. This study provides a foundational understanding of volatile components in the leaves of L. chuanxiong and its medicinal relatives using E-nose combined with HS-SPME-GC-MS, contributing to the discussion on their interspecific odor characteristics and relationships.

Background: In Korea, the medicinal herb Cnidii Rhizoma is manufactured using Cnidium officinale Makino and Ligusticum chuanxiong Hort., with the former being the primary ingredient. C. officinale and L. chuanxiong have similar external appearances, and because their roots are dried for use as medicinal materials, it is difficult to distinguish between them. Therefore, there is a need to develop easy-to-use molecular markers to facilitate the classification of Cnidii Rhizoma varieties and the identification of medicinal materials.BR/Methods and Results: DNA was extracted from C. officinale and L. chuanxiong and analyzed using an Illumina HiSeq 2500 platform. After de novo assembly, the loci showing polymorphisms between C. officinale and L. chuanxiong were identified. Primers were designed for polymorphic loci, and genotyping was performed. Finally, three nuclear genome-based insertion and deletion (InDel) markers, CoInDel-01, CoInDel-02, and CoInDel-03, were developed. These markers produced a single band at 230, 369, and 245 bp, respectively, in C. officinale, whereas two bands were observed in L. chuanxiong at 230 and 264 bp, 304 and 369 bp, and 197 and 245 bp, respectively.BR/Conclusion: The three InDel markers developed in this study will be useful for differentiating between C. officinale and L. chuanxiong

The rhizome of Cnidium officinale Makino (COM) is known for its medicinal properties due to its pharmacological activity. Typically, it is thermally dried after harvesting before being processed into suitable final products. However, because thermal drying affects the levels of thermolabile compounds in COM rhizomes, suitable drying conditions are necessary to ensure the quality of the final product. In this study, we evaluated the effects of drying temperatures (30-70°C) on the levels of pharmacological and antioxidant compounds in COM rhizomes. The highest concentrations of key pharmacological compounds, such as chlorogenic acid, ferulic acid, and ligustilide, were observed at 40°C. While relatively higher levels of total phenolics and flavonoids were also detected at 40°C compared to other drying temperatures, the highest antioxidant activity was observed at room temperature, though it was not significantly different from that at 40°C. These findings suggest that drying COM rhizomes at 40°C is optimal for preserving the levels of pharmacological compounds.

High-throughput sequencing (HTS) was used to comprehensively profile the virome of Cnidium officinale Makino, an economically and medicinally important herb widely cultivated in Korea. Symptomatic leaf samples from nine major cultivation regions were pooled for sequencing. The analysis detected a diverse assemblage of viruses comprising 31 distinct species, including apple stem grooving virus (ASGV), cucumber mosaic virus (CMV), cnidium vein yellowing virus-1 (CnVYV-1), and cnidium virus X (CnVX). Importantly, four novel viral agents were identified: Cnidium officinale virus 1 (CnoV1), cnidium virus Y (CnVY), cnidium virus Z (CnVZ), and a unique cnidium virus–associated satellite RNA (satCnV). Viral read distributions varied markedly by regions; for example, CMV dominated in Bonghwa (two sites) and Geochang, while CnVY prevailed in Samcheok-B and Jecheon. Phylogenetic analyses of ASGV and CMV isolates from C. officinale revealed distinct lineages unique to this host, suggesting host-specific adaptation. Co-infection patterns indicated notable interspecific viral interactions. In particular, we observed a strong antagonism between CnVY and CMV, suggesting competitive exclusion or mutual suppression. A virus co-occurrence network analysis identified additional significant virus–virus associations, and principal component analysis (PCA) of virome profiles showed that samples cluster according to their dominant viruses. Overall, our findings emphasize the power of HTS in uncovering complex viromes and advancing understanding of virus ecology in clonally propagated medicinal crops. The comprehensive virome dataset generated here provides an essential foundation for future epidemiological studies and management strategies in C. officinale and related species.

The extract of the rhizome of Cnidium officinale Makino has potential anti-cancer and anti-inflammatory effects in many diseases, such as cancer. However, the biological functions of falcarindiol (FAD) in breast cancer are not fully understood. This study proved the anti-inflammatory and anti-cancer effects of FAD in breast cancer. Breast cancer models confirmed that FAD reduces cell viability and decreases the tumor volume of xenograft mouse models in a dose-dependent manner. FAD mediated caspase-3-dependent apoptosis in MDA-MB-231 and MCF-7 cells, whereas Z-VAD-FMK in combination with FAD inhibited caspase-3-induced apoptosis. FAD mediates apoptosis through cytosolic reactive oxygen species (ROS) and calcium (Ca2+) production and ER stress signaling pathways. In addition, FAD combined with thapsigargin (TG) exerts a synergistic apoptotic cell death effect. In the loss-of-function experiments, PERK or CHOP ablation suppressed intracellular ROS and Ca2+ release and ER stress-induced apoptosis in FAD-treated breast cancer models. Since there is a relationship between ROS and NADPH Oxidase 4 (NOX4), Nox4 ablation blocked ER stress-mediated apoptotic cell death by inhibiting ROS release in FAD-induced breast cancer models. Radioresistant models, such as MCF-7R and MDA-MB-231R, were developed to address the cellular radioresistance in clinical radiotherapy. FAD combined with radiation (2 Gy) overcame radioresistance via the inhibition of the epithelial-mesenchymal transition (EMT) phenomenon, such as the upregulation of PPARγ, VIM, and CDH2 and the downregulation of CDH1. Consequently, these results show that FAD may be a novel treatment as a breast cancer therapy.

Objectives: This study aimed to investigate wheter Gami-daebo-tang gagambang, commonly used for treating stroke sequelae, induces acute toxicity or genetoxicity.Methods: Gami-daebo-tang gagambang contains that the root of &lt;i&gt;Angelica gigas Nakai&lt;/i&gt; inducing acute headache and weakness, and the rhizome of &lt;i&gt;Cnidium officinale&lt;/i&gt; MAKINO, an herb with potential teratogenicity. Teratogenicity is closely associated with genotoxicity. We analyzed whether Gami-daebo-tang gagambang induces acute toxicity and genotoxicity in accordance with Korean non-clinical test standards and OECD test guidelines TG471, TG473, TG474.Results: When male and female rats were given a single dose of Gami-daebo-tang gagambang, no toxic effects or organ damage were observed at dosages reaching 2,500 mg/kg. The bacterial reverse mutation assay showed no evidence of DNA mutations at concentrations up to 5,000 &lt;i&gt;μ&lt;/i&gt;g/plate. In vitro studies indicated that Gami-daebo-tang gagambang did not cause structural or numerical chromosomal aberrations at concentrations as high as 2,000 &lt;i&gt;μ&lt;/i&gt;g/mL. Moreover, animal studies demonstrated that the substance did not induce bone marrow toxicity or the formation of micronuclei in erythrocytes at doses up to 2,000 mg/kg.Conclusions: The studies conducted with different models showed that Gami-daebo-tang gagambang did not cause acute toxicity or genotoxicity. This study provides evidence for the safety of Gami-daebo-tang gagambang in terms of acute toxicity and genotoxicity, supporting its potential safe application in treating stroke sequelae.

This research was carried out to investigate the characteristics of endophytic bacteria from the medicinal plant Cnidium officinale Makino.The plants were collected from four different fields of Bonghwa, Jeongseon, Jecheon and Taebaek in South Korea.Their endophytic bacteria and DNA were extracted and analyzed.As a result of the culture, eight species of culturable endophytic bacteria could be isolated, and Enterobacter ludwigii and Pantoea agglomerans were identified as the main culturable endophytic bacteria.In addition, next generation sequencing (NGS) analysis was performed to analyze unculturable endophytic bacteria, and five orders including Rhizobiales and Streptomycetales were dominant.As a result of betadiversity analysis, there was no significant difference (p > 0.05) between each region.Three modules were identified through network analysis, and eight connectors with high amongmodule connectivity were confirmed.Among them, the relative frequency of Streptomyces sp. is the highest, and it is considered the most important endophytic bacteria in Cnidium.Function prediction was performed with phylogenetic investigation of communities by reconstruction of unobserved states 2 (PICRUSt 2), and module 1 was highly related to the bacterial secretion system.On the other hand, module 2 and module 3 were highly related to plantpathogen interaction and biosynthesis of siderophore.Therefore, in this study, we suggest that Streptomyces sp.play a key role in maintaining health and growth of Cnidium.

A novel targeted anticancer drug delivery strategy: Cnidium officinale polysaccharide conjugated with carboxymethyl-5-fluorouracil and folic acid for ovarian cancer therapy

Reactive oxygen species (ROS) are produced from energy metabolism and may cause diseases or cell death. Antioxidation refers to the suppression of ROS production and is considered beneficial in preventing diseases. This study aimed to examine the antioxidative effects of Cnidium officinale Makino (COM) extracts and fractions using Caenorhabditis elegans as an experimental model. The COM ethanol extract was fractionated according to polarity. The results showed that the ethyl acetate fraction of COM showed powerful radical scavenging activities and increased the activities of superoxide dismutase (SOD) and catalase in C. elegans in a concentration-dependent manner. Moreover, the ethyl acetate fraction reduced the ROS production rate in C. elegans and increased the cell survival rate, suggesting oxidative and thermal stress resistance. In addition, the SOD-3::green fluorescent protein (GFP) expression level in the transformed cells of C. elegans (CF1553) increased, suggesting oxidative stress resistance. Similarly, the HSP-16.2::GFP expression level increased, suggesting thermal stress resistance. In conclusion, the ethyl acetate fraction of COM demonstrated the strongest antioxidative effects, indicating that it may help extend longevity.

We aimed to assess the antifungal efficacy and impact of a denture cleanser containing Cnidium officinale extract on the surface characteristics of denture base materials, as well as its physical and biological properties. The experimental denture cleansers were formulated with C. officinale at concentrations of 100 and 150 μg/mL, combined with 1% cocamidopropyl betaine as a natural surfactant. Antifungal efficacy was evaluated using zone-of-inhibition assays against Candida albicans, revealing inhibition zones of 20 ± 1.8 mm for the 100 μg/mL concentration and 23.6 ± 1.6 mm for the 150 μg/mL concentration. Surface property assessments-including hardness, roughness, color stability, and solubility measurements-demonstrated no significant differences compared to the control group. Biological evaluations included the quantification of polyphenol and flavonoid content. The C. officinale-based cleanser showed significant antifungal activity without affecting the hardness, roughness, color stability, or solubility of denture base materials. Biological tests revealed no cytotoxicity and minimal mucosal irritation. Polyphenol and flavonoid contents were quantitatively measured, revealing higher concentrations in the experimental groups, which were correlated with significant antifungal activity. These compounds are known for their roles in disrupting microbial processes and enhancing antimicrobial effects. These findings suggest that the C. officinale-based denture cleanser effectively inhibits C. albicans while preserving the physical properties of denture base materials. This study highlights the potential of C. officinale in denture cleanser formulations, promoting denture hygiene and oral health. Future research should prioritize long-term clinical evaluations and formulation optimization.

Objectives: This study aimed to investigate whether Ligigeopoong-san induces acute toxicity and genotoxicityMethods: Ligigeopoong-san contains the rhizome of Cnidium officinale MAKINO, an herb with potential teratogenicity. Teratogenicity is closely associated with genotoxicity. We analyzed whether Ligigeopoong-san induces acute toxicity and genotoxicity using various experimental models in accordance with Korean non-clinical test standards for pharmaceuticals and OECD test guidelines.Results : When Ligigeopoong-san was administered as a single dose to male and female rats, no toxic reactions, including organ damage, were observed at doses up to 2,500 mg/kg. In the bacterial reverse mutation test, no DNA mutations were detected at concentrations up to 5,000 μg/plate. In cell models, Ligigeopoong-san did not induce structural or numerical chromosomal aberrations at concentrations up to 2,000 μg/mL. Additionally, in animal studies, it did not cause bone marrow toxicity or form micronuclei in erythrocytes at doses up to 2,000 mg/kg.Conclusions : The experiments using various models demonstrated that Ligigeopoong-san did not induce acute toxicity or genotoxicity.

HemoHIM is a functional food ingredient comprising a triple herbal combination of extracts from Angelica gigas Nakai, Cnidium officinale Makino, and Paeonia lactiflora Pallas. It was developed to aid the recovery of impaired immune function. Although it is widely used to treat various immune disorders in Korea, its potential toxicity has not been extensively investigated. Therefore, a comprehensive study was conducted to assess the safety of HemoHIM, including acute oral dose toxicity, 28-day and 13-week repeated-dose toxicity, and genotoxicity. To evaluate its safety profile, the dose was increased to 2,000 mg/kg/day, which corresponds to the dose limit for acute toxicity as per the Organization for Economic Cooperation and Development Test Guideline 423. No abnormal findings were observed at the higher doses. For the 28-day and 13-week repeated-dose toxicity studies, HemoHIM was administered at doses of 500, 1,000, and 2,000 mg/kg/day to examine subchronic toxicity in male and female rats. No test item-related clinical signs or mortality was observed at any of the tested doses. Gross pathology, hematology, blood chemistry, and histopathology evaluations further supported the safety of HemoHIM. Therefore, the NOAEL of HemoHIM was considered to be at 2,000 mg/kg/day for both sexes of rats. Bacterial reverse mutation tests, a chromosome aberration test in human peripheral blood lymphocytes, and a mouse micronucleus test were conducted to determine the genotoxicity of HemoHIM, which revealed that HemoHIM was non-mutagenic and non-clastogenic. Collectively, these findings provide valuable evidence to support the safe use of HemoHIM as a functional food ingredient.

Recent studies have focused on using big-data-based machine learning to address the effects of climate change scenarios on the production and quality of medicinal plants. Challenges relating to data collection can hinder the analysis of key feature variables that affect the quality of medicinal plants. In the study presented herein, we analyzed feature variables that affect the phenolic content of Korean Cnidium officinale Makino (C. officinale Makino) under different climate change scenarios. We applied different climate change scenarios based on environmental information obtained from Yeongju city, Gyeongsangbuk-do, Republic of Korea, and cultivated C. officinale Makino to collect data. The collected data included 3237, 75, and 45 records, and data augmentation was performed to address this data imbalance. We designed a function based on the DPPH value to set the phenolic content grade in C. officinale Makino and proposed a stacking ensemble model for predicting the total phenol contents and classifying the phenolic content grades. The regression model in the performance evaluation presented an improvement of 6.23–7.72% in terms of the MAPE; in comparison, the classification model demonstrated a 2.48–3.34% better performance in terms of accuracy. The classification accuracy was &gt;0.825 when classifying phenol content grades using the predicted total phenol content values from the regression model, and the area under the curve values of the model indicated high model fitness (0.987–0.981). We plan to identify the key feature variables for the optimal cultivation of C. officinale Makino and explore the relationships among these feature variables.

Despite extensive studies to identify effective curative drugs for amyotrophic lateral sclerosis (ALS), only riluzole and edaravone have been approved by the Food and Drug Administration. However, these drugs only delay disease progression and exhibit adverse effects, necessitating the development of more effective drugs. Herbal medicines are effective against incurable diseases with various pathogenic factors owing to their low toxicity and presence of multiple components, which target multiple organs. Therefore, we aimed to investigate whether a combined herbal medicine (CHM), comprising Gastrodia elata, Cnidium officinale Makino, and Ostericum koreanum, affects muscle function and motor neuron death in an animal model of ALS. We treated 8-week-old hSOD1G93A mice with 1 mg/g CHM, administered orally once daily for 6 weeks. Muscle function was measured via a footprint test. Biochemical analyses, including immunoblotting, western blotting, and immunohistochemistry, of the muscles (tibialis anterior and gastrocnemius) and spinal cord of hSOD1G93A mice were performed. The CHM treatment improved movement and reduced motor neuron loss in the mouse spinal cord. It also enhanced anti-inflammatory and anti-oxidant activities and regulated autophagy in the mouse muscles and spinal cord. These findings suggest that CHM has multi-active components that effectively target muscles and the spinal cord, delaying disease progression.