1.Exploring Chemical Constituent Distribution in Blood/Brain(Hippocampus) and Emotional Regulatory Effect of Raw and Vinegar-processed Products of Citri Reticulatae Pericarpium Viride
Yi BAO ; Yonggui SONG ; Qianmin LI ; Zhifu AI ; Genhua ZHU ; Ming YANG ; Huanhua XU ; Qin ZHENG ; Yiting HUANG ; Zihan GAO ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):189-197
ObjectiveTo investigate the migration and distribution characteristics of chemical constituents in blood and hippocampal tissues before and after vinegar processing of Citri Reticulatae Pericarpium Viride(CRPV), and to explore the potential material basis and mechanisms underlying their regulatory effects on emotional disorders by comparing the effects of raw and vinegar-processed products of CRPV. MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was employed to characterize and identify the chemical constituents of raw and vinegar-processed products of CRPV extracts, as well as their migrating components in blood and hippocampal tissues after oral administration. Reference standards, databases, and relevant literature were utilized for compound annotation, with data processing performed using PeakView 1.2 software. Seventy male C57BL/6 mice were randomly divided into seven groups, including the blank group, model group, diazepam group(2.5 mg·kg-1), raw CRPV low/high dose groups(0.6, 1.2 g·kg-1), and vinegar-processed CRPV low/high dose groups(0.6, 1.2 g·kg-1), with 10 mice per group. Except for the blank group, all other groups underwent chronic restraint stress(2 h·d-1) for 20 d. Each drug-treated group received oral administration at the predetermined dose starting 10 d after modeling, with a total treatment duration of 10 d. Following model-based drug administration, mice underwent open-field, forced swimming, and elevated plus maze tests. After anesthesia with isoflurane, whole brains were collected from each group of mice, and hippocampi were dissected. Reactive oxygen species(ROS) level in hippocampal tissues was quantified by enzyme-linked immunosorbent assay(ELISA). Hematoxylin-eosin(HE) staining was used to observe hippocampal tissue morphology. Immunofluorescence was performed to detect neuronal nuclei(NeuN) and peroxisome proliferator-activated receptor alpha(PPARα) expressions in hippocampal tissue. Then, pharmacodynamic evaluations were conducted to assess the effects of raw and vinegar-processed CRPV on mood disorders, exploring the potential mechanisms. ResultsVinegar processing caused significant changes in the chemical composition of CRPV, with 18 components showing increased relative content and 35 components showing decreased relative content. The primary changes occurred in flavonoid compounds, including 20 flavonoids, 20 flavonoid glycosides, 3 triterpenes, 3 phenolic acids, 1 alkaloid, and 6 other compounds. Twenty-one components were detected in blood(15 methoxyflavones, 4 flavonoid glycosides, and 2 phenolic acids), with 17 shared between raw and vinegar-processed CRPV. Seven components reached hippocampal tissues(all common to both forms). In regulating emotional disorders, Vinegar-processed CRPV exhibited superior antidepressant-like effects compared to raw products. HE staining revealed that both treatments improved hippocampal neuronal morphology, particularly in the damaged CA1 and CA3 regions. Immunofluorescence and ELISA analyses demonstrated that both raw and vinegar-processed CRPV significantly modulated NeuN and PPARα expressions in hippocampal tissue while alleviating oxidative stress induced by excessive ROS(P<0.05). ConclusionThe chemical composition of CRPV undergoes changes after vinegar processing, but the migrating components in blood and hippocampus are primarily methoxyflavonoids. These components may serve as the potential material basis for activating the PPARα pathway, thereby negatively regulating ROS generation in the hippocampus, reducing oxidative stress, and promoting the development of NeuN-positive neurons. These findings provide experimental evidence for enhancing quality standards, pharmacodynamic material research, and active drug development of raw and vinegar-processed CRPV.
2.Analysis of Differential Metabolites of Pinelliae Rhizoma at Different Browning Stages Based on Widely Targeted Metabolomics
Jing TAO ; Honghong LIANG ; Ruoshi LI ; Zhouli XU ; Minzhao LI ; Aien TAO ; Guihua JIANG ; Li AI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):191-199
ObjectiveTo investigate differential metabolites associated with browning in the post-harvest processing of Pinelliae Rhizoma, providing data support for elucidating the key metabolites and metabolic pathways involved in browning, and developing safe and efficient sulfur-free processing techniques. MethodsUltra-performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry(UPLC-QTRAP-MS/MS) was used to detect the metabolites of Pinelliae Rhizoma at different browning stages(0, 8, 16 h) for widely targeted metabolomics. Subsequently, Multivariate statistical analysis of metabolites was conducted using principal component analysis(PCA), hierarchical cluster analysis(HCA), orthogonal partial least squares-discriminant analysis(OPLS-DA), and K-means cluster analysis. Differential metabolites at different browning stages were screened based on variable importance in the projection(VIP) value>1 and |log2fold change(FC)|≥1, and metabolic pathway enrichment analysis was performed using Kyoto Encyclopedia of Genes and Genomes(KEGG). ResultsA total of 1 416 metabolites were identified across the three browning stages of Pinelliae Rhizoma, predominantly comprising amino acids and their derivatives(239), lipids(219), alkaloids(156), phenolic acids(121), terpenoids(113), and flavonoids(111). A two-by-two comparison of the three browning phases, yielded 622 differential metabolites that were significantly enriched in the phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and purine metabolic pathway. Further analysis revealed that carbohydrates such as D-mannose and turanose, phenolic acids such as 1-O-caffeoyl-6-O-glucosyl-β-D-glucose, dicaffeoylshikimic acid, and flavonoids such as epigallocatechin gallate, vitexin-7-O-rutinoside, luteolin-7-O-(6″-malonyl)glucoside-5-O-arabinoside, catechin gallate, epicatechin gallate, isovitexin-7-O-glucoside-2″-O-rhamnoside, apigenin-7-O-rutinoside-4ʹ-O-sophoroside, 3,5,3ʹ,4ʹ,5ʹ-penta-hydroxyflavan-7-gallate may act as browning substrates and play important roles in the browning process. ConclusionCarbohydrates, phenolic acids, and flavonoids may serve as key substrates in the browning process of Pinelliae Rhizoma, involving pathways such as phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and purine metabolism, which can provide a theoretical basis for further exploration of the browning mechanism.
3.Influencing Factors of Urate Crystal Deposition in Patients with Hyperuricemia and Prediction Model of TCM Syndrome Types-inflammatory Indicators
Jiaqi XU ; Bin AI ; Chao LIN ; Qiaoxuan LIN ; Changning LI ; Jing CAI ; Yan XIAO ; Jiemei GUO ; Youxin SU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):66-73
ObjectiveTo identify potential influencing factors of urate crystal deposition at ankle/foot in patients with hyperuricemia (HUA), and to analyze the predictive value of inflammatory indicators for urate crystal deposition in patients with different traditional Chinese medicine (TCM) syndromes, so as to provide potential reference for clinical risk assessment and individualized TCM intervention. MethodsA retrospective study was carried out with the enrollment of 231 HUA patients from The Third Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine between January 2021 and December 2024. The enrolled patients were further divided into a crystal deposition-positive group (143 cases) and a crystal deposition-negative group (88 cases) according to the results of dual-energy computed tomography (CT). Sociodemographic data, living habits, serum uric acid levels, and inflammatory indicators of the enrolled patients were collcted, and TCM syndrome differentiation was performed. Furthermore, univariate analysis was used to compare inter-group differences in clinical characteristics. MMultivariate Logistic regression was applied to identify the influencing factors of urate crystal deposition. In addition, the receiver operating characteristic (ROC) curves were plotted to evaluate the predictive efficacy of inflammatory indicators for crystal deposition across different TCM syndromes. ResultsThere were statistically significant inter-group differences in the proportion of males, age, body mass index, proportion of mental labor, rate of low water intake, and rate of high-sugar beverage consumption (P<0.05),whereas no significant difference in low exercise intensity was found between the two groups. Furthermore, compared with the negative group, the positive group had higher serum uric acid level, neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR), but lower systemic immune-inflammation index (SIRI) (P<0.05). Regarding the distribution of TCM syndromes, the positive group was dominated by the dampness-heat accumulation syndrome (55/143,38.46%), while the negative group was mainly characterized by the phlegm-turbidity obstruction syndrome (44/88,50.00%). Multivariate Logistic regression analysis revealed that high-sugar beverage consumption, elevated NLR, and elevated PLR were risk factors for urate crystal deposition [odd ratio (OR) = 8.002, 5.377, 1.034, respectively; 95% CI 1.572-40.732, 2.179-13.270, 1.013-1.054,all P<0.05], while SIRI was a protective factor (OR = 0.869, 95% CI 0.778-0.971, P<0.05). In the positive group, patients with the dampness-heat accumulation syndrome exhibited the highest NLR, while the lowest PLR and SIRI, showing statistically significant differences with those of other syndromes (all P<0.05). In addition, ROC curve analysis indicated that for the dampness-heat accumulation syndrome, the combined "NLR + PLR" model had an area under the curve (AUC) of 0.901 (95% CI 0.850-0.951, P<0.01), with a sensitivity of 89.1% and a specificity of 79.5%; for the blood stasis-heat obstruction syndrome, the combined "NLR + PLR" model had an AUC of 0.880 (95% CI 0.825-0.934, P<0.01), with a sensitivity of 100.0% and a specificity of 67.3%; for the liver-kidney Yin-deficiency syndrome, the single PLR model had an AUC of 0.842 (95% CI 0.731-0.952, P<0.01), with a sensitivity of 83.3% and a specificity of 84.0%. ConclusionUrate crystal deposition in HUA patients exhibits intimate associations with high-sugar beverage consumption as well as elevated NLR and PLR levels. Meanwhile, TCM syndrome differentiation has potential correlation with inflammatory characteristics. The inflammatory indicator-based prediction model constructed based on TCM syndromes exhibits good predictive value.
4.Exploring Therapeutic Effect of Yuejuwan on Depressed Mice Based on Lipidomics
Zhentao ZHANG ; Dan SU ; Huizhen LI ; Yonggui SONG ; Huanhua XU ; Meixizi LAI ; Zhifu AI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):85-94
ObjectiveTo investigate the effect of Yuejuwan on lipid metabolism in serum, prefrontal cortex and hippocampus of depressed mice based on lipidomics, and to explore the potential pathways for improving lipid metabolism to prevent depression. MethodsSeven-week-old C57BL/6 mice were randomly divided into blank group, model group, Yuejuwan group(3.6 g·kg-1) and fluoxetine group(10 mg·kg-1), and chronic unpredictable mild stress(CUMS) was used to establish the depression model. After 3 weeks of modeling, each administration group was gavaged with the corresponding drug solution according to the dose, and mice in the blank and model groups were given an equal volume of deionised water by gavage, one time/d for 2 weeks. After administration, the antidepressant effect of Yuejuwan was evaluated by neurobehavioral indices such as sucrose preference test, open field test, tail suspension test and forced swimming test. An automatic biochemical analyzer was used to measure contents of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in mouse serum. Lipidomic analysis of mouse serum, prefrontal cortex and hippocampus was performed based on ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UPLC-LTQ-Orbitrap-MS), and the expression of mammalian target of rapamycin(mTOR), ribosomal protein S6 kinase(S6K), phosphorylation(p)-mTOR, p-S6K in gastric tissues of mice was detected by Western blot. ResultsCompared with the blank group, mice in the model group exhibited significantly reduced sucrose preference rate and center movement time in the open field test(P<0.01), the immobility times in the tail suspension test and forced swimming test were significantly increased(P<0.01), and serum levels of TC, TG, LDL-C, HDL-C, AST and ALT were significantly elevated(P<0.05, P<0.01). Compared with the model group, the Yuejuwan group showed a significant increase in the sucrose preference rate and center movement time in the open field test(P<0.01), the immobility times in the tail suspension test and forced swimming test were significantly reduced(P<0.01), and the serum levels of TC, TG, LDL-C, AST and ALT were significantly decreased(P<0.05, P<0.01). Lipidomic analysis revealed that Yuejuwan had a significant effect on lipid metabolism in serum, prefrontal cortex and hippocampus of depressed mice, and The differential lipid metabolites were mainly enriched in the metabolic pathways of glycerophospholipid metabolism, sphingolipid signaling, and glycosylphosphatidylinositol-anchored protein biosynthesis, among which the glycerophospholipid metabolic pathway was the most significant. Western blot results showed that compared with the blank group, the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the model group were significantly increased(P<0.01). In comparison with the model group, the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the Yuejuwan group were significantly decreased(P<0.01). ConclusionThe intervention of Yuejuwan on lipid metabolism is one of the potential pathways for its antidepressant effect, which may be related to the regulation of mTOR/S6K signaling pathway upstream of lipid metabolism in the gastric tissues.
5.Effects and mechanism of asperuloside on the pyroptosis of intestinal epithelial cells in rats with ulcerative colitis
Chao XU ; Xiaoping TAN ; Jie LI ; Minghua AI ; Yueyue LU ; Chaoyong LIU
China Pharmacy 2025;36(2):166-171
OBJECTIVE To investigate the effects and mechanism of asperuloside (Asp) on the pyroptosis of intestinal epithelial cells in rats with ulcerative colitis (UC). METHODS The male SD rats were randomly divided into Control group, model group (UC group), ASP low-dose and high-dose groups [Asp-L, Asp-H groups, Asp 35, 70 mg/(kg·d)], ASP high-dose group+AMPK inhibitor Compound C group [Asp-H+Compound C group, Asp 70 mg/(kg·d)+Compound C 0.2 mg/(kg·d)], with 12 rats in each group. Except for Control group, the other groups were injected with 50% ethanol (0.25 mL)+5% 2,4, 6- trinitrobenzene sulfonic acid solution (2 mL/kg) into the intestinal cavity to construct UC model. After modeling, the rats in each drug group were given corresponding drug solution by gavage or (and) tail vein injection, once a day, for 14 consecutive days. After the last administration, the weight of rats in each group was measured, and the length of their colons was measured; disease activity index (DAI) score and colonic mucosal damage index (CMDI) score were performed, and the serum levels of inflammatory factors (interleukin-18, -1β, -6) were detected. The pathological changes of the colon tissue were observed. The expressions of pyroptosis-related proteins [caspase-1, gasdermin D (GSDMD)] in colon tissue, and pathway-related proteins such as adenosine monophosphate-activated protein kinase (AMPK), thioredoxin-interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3) and apoptosis-associated speck-like protein containing a CARD (ASC) were all detected. RESULTS Compared with Control group, the colon tissue structure of rats in UC group was damaged, with obvious infiltration of inflammatory cells and edema. Their body weight, colon length and phosphorylation level of AMPK protein were significantly reduced or shortened; DAI and CMDI scores, serum levels of inflammatory factors, and the protein expressions of caspase-1, GSDMD, TXNIP, NLRP3 and ASC in colon tissue were increased or upregulated significantly (P<0.05). Compared with UC group, the pathological damage of colon tissue in rats was relieved in Asp-L and Asp-H groups, and all quantitative indicators were significantly improved (P<0.05); the improvement effect of Asp-H group was more significant (P<0.05). Compound C could significantly reverse the improvement effect of high-dose of Asp on the above indicators in UC rats (P<0.05). CONCLUSIONS Asp can improve inflammatory damage in colon tissue and inhibit pyroptosis of intestinal epithelial cells in UC rats, which is associated with the activation of AMPK and inhibition of TXNIP/NLRP3 signaling pathway.
6.Effects and mechanism of asperuloside on the pyroptosis of intestinal epithelial cells in rats with ulcerative colitis
Chao XU ; Xiaoping TAN ; Jie LI ; Minghua AI ; Yueyue LU ; Chaoyong LIU
China Pharmacy 2025;36(2):166-171
OBJECTIVE To investigate the effects and mechanism of asperuloside (Asp) on the pyroptosis of intestinal epithelial cells in rats with ulcerative colitis (UC). METHODS The male SD rats were randomly divided into Control group, model group (UC group), ASP low-dose and high-dose groups [Asp-L, Asp-H groups, Asp 35, 70 mg/(kg·d)], ASP high-dose group+AMPK inhibitor Compound C group [Asp-H+Compound C group, Asp 70 mg/(kg·d)+Compound C 0.2 mg/(kg·d)], with 12 rats in each group. Except for Control group, the other groups were injected with 50% ethanol (0.25 mL)+5% 2,4, 6- trinitrobenzene sulfonic acid solution (2 mL/kg) into the intestinal cavity to construct UC model. After modeling, the rats in each drug group were given corresponding drug solution by gavage or (and) tail vein injection, once a day, for 14 consecutive days. After the last administration, the weight of rats in each group was measured, and the length of their colons was measured; disease activity index (DAI) score and colonic mucosal damage index (CMDI) score were performed, and the serum levels of inflammatory factors (interleukin-18, -1β, -6) were detected. The pathological changes of the colon tissue were observed. The expressions of pyroptosis-related proteins [caspase-1, gasdermin D (GSDMD)] in colon tissue, and pathway-related proteins such as adenosine monophosphate-activated protein kinase (AMPK), thioredoxin-interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3) and apoptosis-associated speck-like protein containing a CARD (ASC) were all detected. RESULTS Compared with Control group, the colon tissue structure of rats in UC group was damaged, with obvious infiltration of inflammatory cells and edema. Their body weight, colon length and phosphorylation level of AMPK protein were significantly reduced or shortened; DAI and CMDI scores, serum levels of inflammatory factors, and the protein expressions of caspase-1, GSDMD, TXNIP, NLRP3 and ASC in colon tissue were increased or upregulated significantly (P<0.05). Compared with UC group, the pathological damage of colon tissue in rats was relieved in Asp-L and Asp-H groups, and all quantitative indicators were significantly improved (P<0.05); the improvement effect of Asp-H group was more significant (P<0.05). Compound C could significantly reverse the improvement effect of high-dose of Asp on the above indicators in UC rats (P<0.05). CONCLUSIONS Asp can improve inflammatory damage in colon tissue and inhibit pyroptosis of intestinal epithelial cells in UC rats, which is associated with the activation of AMPK and inhibition of TXNIP/NLRP3 signaling pathway.
7.Circular RNA hsa_circ_0081621 promotes malignant biological behavior of human laryngeal squamous cell carcinoma cells
LIU Shenghui1,2 ; AI Xueying1,2 ; XU Yuru1 ; LI Shuqi1,2 ; LIU Meng1 ; HU Guobin1 ; SHAN Baoen2,3
Chinese Journal of Cancer Biotherapy 2025;32(6):579-586
[摘 要] 目的:探究环状RNA hsa_circ_0081621对人喉鳞状细胞癌AMC-HN-8和TU177细胞恶性生物学行为的影响。方法:常规培养AMC-HN-8和TU177细胞,用转染试剂将si-NC、si-hsa_circ_0081621、空载体(vector)和hsa_circ_0081621过表达载体(hsa_circ_0081621-OE)转染至AMC-HN-8和TU177细胞,分别记作si-NC、si-hsa_circ_0081621、vector和hsa_circ_0081621-OE组。用CCK-8法、克隆形成实验、划痕愈合实验和Transwell小室实验分别检测敲减或过表达hsa_circ_0081621对AMC-HN-8和TU177细胞增殖、迁移和侵袭能力的影响。结果:在AMC-HN-8和TU177中成功地敲减或过表达了hsa_circ_0081621;敲减或过表达hsa_circ_0081621能显著抑制或促进AMC-HN-8和TU177细胞的增殖、迁移和侵袭能力(P < 0.01或P < 0.001或P < 0.000 1)。结论:hsa_circ_0081621可促进人喉鳞状细胞癌AMC-HN-8和TU177细胞的恶性生物学行为。
8.Adhesion Mechanisms of Aquatic Fouling Organisms Mediated by Biomacromolecules
Dan HE ; Shi-Guo LI ; Ai-Bin ZHAN
Progress in Biochemistry and Biophysics 2025;52(7):1833-1852
Aquatic organisms can secrete biomacromolecules through specialized organs, tissues, or structures, enabling adhesion to underwater material surfaces and leading to severe biofouling issues. This phenomenon adversely impacts aquatic ecosystem health and human activities. Biofouling has emerged as an emerging global environmental challenge. Adhesion serves as the foundation of biofouling, representing a critical step toward a comprehensive understanding of the adhesion mechanisms of aquatic organisms. Biomacromolecules, including proteins, lipids, and carbohydrates, are the primary functional components in the adhesive substances of aquatic fouling organisms. Research indicates that these biomacromolecules exhibit diversity in types and characteristics across different aquatic organisms, yet their adhesion mechanisms show unifying features. Despite significant progress, there remains a lack of comprehensive reviews on the adhesion mechanisms mediated by biomacromolecules in aquatic fouling organisms, particularly on the roles of lipids and carbohydrates. Through a comprehensive analysis of existing literature, this review systematically summarizes the mechanistic roles of three classes of macromolecules in aquatic biofouling adhesion processes. Proteins demonstrate central functionality in interfacial adhesion and cohesion through specialized functional amino acids, conserved structural domains, and post-translational modifications. Lipids enhance structural stability via hydrophobic barrier formation and antioxidative protection mechanisms. Carbohydrates contribute to adhesion persistence through cohesive reinforcement and enzymatic resistance of adhesive matrices. Building upon these mechanisms, this review proposes four prospective research directions: optimization of protein-mediated adhesion functionality, elucidation of lipid participation in adhesion dynamics, systematic characterization of carbohydrate adhesion modalities, and investigation of macromolecular synergy in composite adhesive systems. The synthesized knowledge provides critical insights into underwater adhesion mechanisms of aquatic fouling organisms and establishes a theoretical foundation for developing mechanism-driven antifouling strategies. This work advances fundamental understanding of bioadhesion phenomena while offering practical guidance for next-generation antifouling technology development.
9.Influencing factors of treatment compliance in elderly advanced gastric cancer with cancer pain
Tengyue AI ; Zhuqing ZHANG ; Panpan LI ; Zhen NI ; Hongyong LI
Journal of Public Health and Preventive Medicine 2025;36(4):161-164
Objective To analyze the status quo and related factors of treatment compliance in elderly patients with advanced gastric cancer complicated with cancer pain. Methods Three hundred and ten elderly patients with advanced gastric cancer from January 2021 to June 2024 were selected, and their general data were collected. Hospital Anxiety and Depression Scale (HADS), Beliefs about Medicines Questionnaire (BMQ), Numerical Rating Scale (NRS) and 8-item Morisky Medication Adherence Scale (MMAS-8) were adopted to investigate the psychological status, necessity of medication belief, degree of cancer pain and treatment compliance. Univariate analysis, correlation analysis and multivariate logistic regression analysis were used to describe and analyze the treatment compliance status and related factors in elderly advanced gastric cancer with cancer pain. Results A total of 310 patients were divided into group A (good compliance, 93 cases), group B (moderate compliance ,102 cases) and group C (poor compliance , 115 cases) by means of compliance degree. There were significant differences in education level , pain duration and intensity and analgesic category (P<0.05). Treatment compliance was associated with education level, analgesic category, psychological status, medication belief and pain degree (P<0.05). The necessity of medication belief and pain duration and degree were influencing factors of treatment compliance. Conclusion The treatment compliance in advanced gastric cancer with cancer pain is poor, and can be affected by the necessity of medication belief and duration and degree of pain.
10.Material basis of bitter taste and taste-effect relationship in Cistanche deserticola based on UPLC-Q-Orbitrap HRMS combined with molecular docking.
Li-Ying TIAN ; Ming-Jie LI ; Qiang HOU ; Zheng-Yuan WANG ; Ai-Sai-Ti GULIZIYE ; Jun-Ping HU
China Journal of Chinese Materia Medica 2025;50(6):1569-1580
Based on ultra-performance liquid chromatography-quadrupole-electrostatic field Orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap HRMS) technology and molecular docking, the bitter-tasting substances(hereafter referred to as "bitter substances") in Cistanche deserticola extract were investigated, and the bitter taste and efficacy relationship was explored to lay the foundation for future research on de-bittering and taste correction. Firstly, UPLC-Q-Orbitrap HRMS was used for the qualitative analysis of the constituents of C. deserticola, and 69 chemical components were identified. These chemical components were then subjected to molecular docking with the bitter taste receptor, leading to the screening of 20 bitter substances, including 6 phenylethanol glycosides, 5 flavonoids, 3 phenolic acids, 2 cycloalkenyl ether terpenes, 2 alkaloids, and 2 other components. Nine batches of fresh C. deserticola samples were collected from the same origin but harvested at different months. These samples were divided into groups based on harvest month and plant part. The bitterness was quantified using an electronic tongue, and the content of six potential bitter-active compounds(pineconotyloside, trichothecene glycoside, tubulin A, iso-trichothecene glycoside, jinshihuaoside, and jingnipinoside) was determined by high-performance liquid chromatography(HPLC). The total content of phenylethanol glycosides, polysaccharides, alkaloids, flavonoids, and phenolic acids was determined using UV-visible spectrophotometry. Chemometric analyses were then conducted, including Pearson's correlation analysis, gray correlation analysis, and orthogonal partial least squares discriminant analysis(OPLS-DA), to identify the bitter components in C. deserticola. The results were consistent with the molecular docking findings, and the two methods mutually supported each other. Finally, network pharmacological predictions and analyses were performed to explore the relationship between the targets of bitter substances and their efficacy. The results indicated that key targets of the bitter substances included EGFR, PIK3CB, and PTK2. These substances may exert their bitter effects by acting on relevant disease targets, confirming that the bitter substances in C. deserticola are the material basis of its bitter taste efficacy. In conclusion, this study suggests that the phenylethanol glycosides, primarily pineconotyloside, mauritiana glycoside, and gibberellin, are the material basis for the "bitter taste" of C. deserticola. The molecular docking technique plays a guiding role in the screening of bitter substances in traditional Chinese medicine(TCM). The bitter substances in C. deserticola not only contribute to its bitter taste but also support the concept of the "taste-efficacy" relationship in TCM, providing valuable insights and references for future research in this area.
Molecular Docking Simulation
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Taste
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Chromatography, High Pressure Liquid
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Cistanche/chemistry*
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Drugs, Chinese Herbal/chemistry*
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Humans
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Mass Spectrometry


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