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.Treatment of Alzheimer's Disease with Traditional Chinese Medicine: A Review
Zheng XU ; Yuan TANG ; Fenglan QIU ; Yiguang LI ; Lingyu YANG ; Jie CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):322-330
Alzheimer's disease (AD) is a common type of dementia, primarily characterized by cognitive and behavioral impairments as well as deficits in learning and memory. The progression of AD has imposed a significant economic burden on society and families. However, its exact pathogenesis has not yet been fully elucidated. Currently, available therapeutic drugs are limited and are often accompanied by serious adverse effects. Traditional Chinese medicines (TCMs) and their extracts are mostly natural products and possess advantages such as multi-pathway regulation and relatively few adverse reactions. Experimental studies have shown that TCMs exhibit great potential in the prevention and treatment of AD. For example, Huanglian Jieduang, Danggui Shaoyaosan, Kaixin San, Liuwei Dihuangwan, Buyang Huanwutang, as well as Ginseng Radix et Rhizoma, Astragali Radix, Uncariae Ramulus cum Uncis, Coptidis Rhizoma, Gardeniae Fructus, Ginkgo Folium, Salviae Miltiorrhizae Radix et Rhizoma, and Curcumae Longae Rhizoma, can reduce β-amyloid deposition, inhibit excessive Tau protein phosphorylation, restore mitochondrial function, alleviate oxidative stress, suppress neuroinflammation and apoptosis, repair synaptic function, and improve gut microbiota. This article mainly summarizes the effects of several TCMs and compound prescriptions on AD, aiming to provide a reference for subsequent TCM-based treatment of AD.
3.Exploring Anti-inflammatory Synergistic Mechanism of Atractylodis Macrocephalae Rhizoma Processed with Aurantii Fructus Immaturus Juice Based on Differential Component Tracking Strategy
Hongda XUAN ; Shengnan SHEN ; Linlin LI ; Jingjing LIAO ; Xianyu XU ; Xiaoxia LIU ; Haining LYU ; Fang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):228-237
ObjectiveTaking Aurantii Fructus Immaturus juice(AFI)-processed Atractylodis Macrocephalae Rhizoma(AMR) as an example, this study aims to systematically compare the volatile and non-volatile components of AMR and its processed products, investigate the key differential components, evaluate their anti-inflammatory activities, and elucidate the synergistic mechanism of processing. MethodsThe chemical compositions of volatile and non-volatile components in AMR and AFI-processed AMR were systematically characterized using gas chromatography-mass spectrometry(GC-MS) and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), with relative mass fractions and response values determined separately. Volatile components were identified through searches in the National Institute of Standards and Technology(NIST)17 database, comparison with retention index(RI) and fragmentation pattern matching. Non-volatile components were identified by searching Waters Traditional Chinese Medicine (TCM) spectral library, in conjunction with PubChem and MassBank, characteristic fragmentation patterns and response values were also used to support identification. Differential components were screened using principal component analysis(PCA), orthogonal partial least squares-discriminant analysis(OPLS-DA), with variable importance in the projection(VIP) value >1. Components with high log2fold change(FC) among major differential groups were selected as those exhibiting significant changes before and after processing. The anti-inflammatory activity of the differential compounds was evaluated by assessing their effects on nitric oxide(NO) production in a lipopolysaccharide(LPS)-induced RAW264.7 macrophage model. Enzyme-linked immunosorbent assay(ELISA) was used to detect the effects of the differential components on tumor necrosis factor(TNF)-α, interleukin(IL)-1β, IL-6, and monocyte chemotactic protein(MCP)-1 levels, and immunofluorescence(IF) was employed to assess their effects on nuclear transcription factor(NF)-κB p65 translocation, thereby elucidating the underlying molecular mechanisms. ResultsA total of 36 compounds were identified in the volatile components of AMR and AFI-processed AMR, among which, sesquiterpenes and monoterpenes were significantly increased after processing. In the non-volatile components, 36 compounds were identified, and the main differential components were flavonoids, sesquiterpenoids, and triterpenoids. Flavonoids were the primary differential components distinguishing AMR from its processed products, representing compounds directly introduced during processing. Five compounds, including atractylenolide Ⅲ, tangeritin, nobiletin, hesperidin and narirutin, were selected as representatives of three classes based on their most prominent differential expression among different compound types for subsequent anti-inflammatory activity studies. The results showed that 100 μmol·L-1 tangerine and narirutin could significantly inhibit LPS-induced NO production(P<0.01) in a concentration-dependent manner. Tangeritin was able to significantly inhibit the levels of TNF-α and MCP-1 secreted by RAW264.7(P<0.05), while narirutin significantly inhibited the levels of TNF-α, IL-1β, MCP-1 and IL-6(P<0.01). IF revealed that both tangeritin and narirutin significantly blocked the translocation of NF-κB p65 from the cytoplasm to the nucleus. ConclusionAFI-processed AMR significantly alters the chemical composition profile of AMR, and the newly introduced flavonoid components during processing may be key to its enhanced anti-inflammatory effects.
4.Mechanism of action of Qingre huatan huoxue decoction against atherosclerosis based on macrophage polarization
Huaping ZHONG ; Qicheng ZHU ; Zhengwei ZOU ; Zhengyi HE ; Heping XIE ; Xu CHEN ; Zhisheng DUAN ; Tian XIAO
China Pharmacy 2026;37(4):438-443
OBJECTIVE To explore the mechanism of action of Qingre huatan huoxue decoction against atherosclerosis (AS)based on macrophage polarization. METHODS Using atorvastatin served as the positive control, the drug-containing serum of the Qingre huatan huoxue decoction was prepared to treat RAW264.7 macrophages. Macrophage viability, apoptosis rate, and the fluorescence intensities of CD86 and CD206 were measured, along with the levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). Apolipoprotei n E-deficient (ApoE -/- ) mice (AS model mice) fed with a high-fat diet were randomly assigned to model group, atorvastatin group (2.6 mg/kg), and low-, medium- and high-dose groups of Qingre huatan huoxue decoction (90, 180, 360 mg/kg), respectively. C57BL/6J mice fed with a standard diet served as the normal control group, with 10 mice per group. The treatment group mice were administered the corresponding drugs intragastrically, once daily, for 8 consecutive weeks. Serum levels of TNF-α and IL-1β were measured in all groups. Lipid deposition in the aorta (assessed by the percentage of plaque in the entire aorta and aortic root) and morphological changes in the aortic root were observed. Expression levels of CD86 and CD206 in aortic tissue, as well as the protein expression levels of inducible nitric oxide synthase (iNOS), arginase-1 (Arg-1), AMP-activated protein kinase (AMPK), phosphorylated AMPK (p-AMPK), and peroxisome proliferator-activated receptor γ (PPAR-γ) in aortic tissues were all detected. RESULTS Cell experiment results showed that, at concentrations of 5-100 μg/mL, the drug-containing serum of the Qingre huatan huoxue decoction significantly increased RAW264.7 cell viability ( P <0.05). The drug-containing serum of the Qingre huatan huoxue decoction at concentrations of 10, 50, and 100 μg/mL, along with atorvastatin, significantly reduced apoptosis rates, CD86 fluorescence intensity, and TNF-α and IL-1β levels in RAW264.7 cells, while markedly enhancing CD206 fluorescence intensity ( P <0.05). Animal experiment results showed that, compared with the model group, all dosage groups of Qingre huatan huoxue decoction and the atorvastatin group showed significantly reduced/down-regulated levels of TNF-α and IL-1β in serum, along with decreased aortic total and root plaque percentages, CD86 expression, and iNOS protein expression. CD206 expression and Arg-1, p-AMPK/AMPK, PPAR-γ protein expression were significantly up-regulated ( P <0.05). Pathological morphology of the aorta showed varying degrees of improvement. CONCLUSIONS The formula of Qingre huatan huoxue decoction exerts its anti-AS effects by regulating macrophage polarization, increasing the proportion of M2 macrophages, thereby effectively inhibiting AS plaque formation and reducing inflammatory responses.
5.Mechanism of Yangjing Zhongyutang in Regulating SIRT1/PGC-1α Signaling Pathway to Promote Mitochondrial Function and Alleviate Oxidative Stress Damage in Rats with Diminished Ovarian Reserve
Ping ZHANG ; Lijuan YANG ; Shenghui CHEN ; Wenliang YAO ; Yuliang ZHOU ; Ling MA ; Huiying WU ; Yanwen XU ; Ziyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):46-55
ObjectiveTo observe the effects of Yangjing Zhongyutang (YJZYT) on mitochondrial biogenesis and oxidative stress damage mediated by the silent information regulator 1 (SIRT1)/peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α) signaling pathway in cyclophosphamide (CTX)-induced rats with diminished ovarian reserve (DOR), and to explore its mechanism in improving ovarian reserve function and follicular development. MethodsForty-two 8-week-old female SD rats with normal estrous cycles were randomly divided into a blank control group (n=7) and a model group (n=35). Rats in the model group received a single intraperitoneal injection of CTX (90 mg·kg-1) to establish the DOR model. After modeling, estrous cycles were monitored for 7 consecutive days, and model success was confirmed based on criteria for estrous cycle disruption. After successful modeling, rats were divided into groups for intervention: estradiol valerate group (0.09 mg·kg-1), and YJZYT high-, medium-, and low-dose groups (19.98, 9.99, 5.00 g·kg-1). The blank control group and model group were given an equal volume of distilled water by gavage. All groups received daily gavage once for 4 consecutive weeks. The general state, body weight, and ovarian wet weight of rats were observed and recorded, and the ovarian organ index was calculated. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). Hematoxylin-eosin (HE) staining was performed to observe ovarian histomorphological changes and follicular development status. Immunofluorescence was used to detect reactive oxygen species (ROS) expression levels. Colorimetric assays were employed to measure adenosine triphosphate (ATP) and malondialdehyde (MDA) content in ovarian tissues. Quantitative Real-time polymerase chain reaction (Real-time PCR) was used to detect mitochondrial DNA (mtDNA) copy number and the mRNA expression levels of key genes including SIRT1, PGC-1α, nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (TFAM). Western blot was performed to detect the protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM. ResultsCompared with the blank group, rats in the model group exhibited disrupted estrous cycles, obviously reduced body weight, and decreased ovarian index (P<0.05). Ovarian histopathology revealed cortical thinning, loose structure, and a significant reduction in both primordial and growing follicles (P<0.01). Serum FSH and LH levels were significantly elevated (P<0.01), while E2 and AMH levels were obviously reduced (P<0.05, P<0.01). ATP content and mtDNA copy number decreased in ovarian tissue (P<0.01), ROS expression increased, MDA levels rose, while SOD and GSH-Px activities obviously decreased (P<0.05, P<0.01), mRNA and protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM were obviously downregulated (P<0.05, P<0.01). After treatment, compared with the model group, body weight and ovarian index obviously recovered in rats administered various doses of YJZYT (P<0.05), serum E2 and AMH levels increased, while FSH and LH levels obviously decreased (P<0.05, P<0.01), ovarian tissue ATP content and mtDNA copy number were up-regulated, ROS and MDA levels decreased, and antioxidant enzymes SOD and GSH-Px activity obviously increased (P<0.05, P<0.01), Gene and protein expression levels related to the SIRT1/PGC-1α /NRF1/TFAM signaling pathway were obviously up-regulated compared to the model group (P<0.05, P<0.01), HE staining revealed that ovarian structure gradually recovered to integrity in all treatment groups, with a obviously increase in the number of primordial and growing follicles (P<0.05, P<0.01). Granulosa cells were neatly arranged, indicating marked improvement in ovarian function. ConclusionYJZYT may improve ovarian function and follicular development in rats with diminished ovarian reserve by activating the SIRT1/PGC-1α signaling pathway, promoting mitochondrial biogenesis, enhancing mitochondrial function, and alleviating oxidative stress damage.
6.Correlation between hand foot mouth disease incidence and meteorological factors with assessment for excess incidence risk among children in Jiujiang City
XIE Wenjing, XU Ting, ZHANG Jingjing, LI Jie, FU Weijie
Chinese Journal of School Health 2026;47(3):417-420
Objective:
To discuss the epidemiological characteristics of hand foot mouth disease (HFMD) among children in Jiujiang City, and its correlation with meteorological factors, so as to provide scientific evidence for developing targeted HFMD prevention and control measures.
Methods:
HFMD incidence data among children and meteorological factor records from Jiujiang City during 2019-2023 were collected. Pearson correlation analysis and Quasi-Poisson regression analysis of generalized additive model were used to analyze the relationship between the number of cases and meteorological factors, calculating correlation coefficients (r) and excess risk (ER).
Results:
The annual reported children HFMD cases in Jiujiang City during 2019-2023 were 4 299, 4 671, 2 560, 3 230, and 3 584, respectively. Except for a peak in autumn-winter in 2020, outbreaks occurred mainly in spring and summer in other years. The majority of cases involved scattered children (71.11%) and children in daycare centers (26.64%). Pearson correlation analysis showed that total case counts, cases among scattered children, and cases among children in daycare centers were positively correlated with average temperature ( r =0.36, 0.39, 0.23), maximum temperature ( r =0.32, 0.36, 0.20), minimum temperature ( r =0.37, 0.41, 0.24), and average relative humidity ( r =0.19, 0.20, 0.15) (all P <0.05). Quasi-Poisson regression analysis of generalized additive model revealed that total cases, cases among scattered children, and cases among daycare children were positively associated with average temperature ( r =0.05, 0.06, 0.03) and average relative humidity ( r =0.03, 0.02, 0.02) (all P <0.01). Excess incidence risk analysis indicated that for every 10 ℃ increase in average temperature, the ER (95% CI ) values were 72.16 (45.91-103.14), 79.76 (52.99- 111.23 ) and 39.30 (14.45-69.54) among total children, scattered children and daycare children, respectively; for every 10% increase in average relative humidity, the ER (95% CI ) values were 28.69 (12.22-47.56), 27.81 (12.01-45.84) and 24.42 (5.56-46.65), respectively.
Conclusions
The incidence level of HFMD among children in Jiujiang City is positively correlated with the average temperature and average relative humidity. Higher temperature and humidity are associated with an increased risk of excess HFMD incidence in scattered children compared to children in daycare centers.
7.Herbal Textual Research on Bambusae Succus in Famous Classical Formulas
Yu SHI ; Feng ZHOU ; Yihan WANG ; Yanmeng LIU ; Ming YANG ; Zhiping CHEN ; Jiangshan ZHANG ; Conglong XU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):231-239
This article systematically reviews and examines the historical evolution of Bambusae Succus as a medicinal material, covering aspects such as nomenclature, origin, geographical distribution, harvesting and processing methods, quality assessment, therapeutic effects and indications, by consulting ancient herbal texts, medical compendia, and modern literature. The aim is to provide a reference for the development and utilization of famous classical formulas containing this herb. Research indicated that Bambusae Succus was first documented in the Shennong Bencaojing during the Han dynasty, with Zhuli being the standard name used throughout history, alongside aliases like Zhuzhi, Zhuyou and Huoquan. Historically, the primary source of Bambusae Succus has been Phyllostachys nigra var. henonis(Danzhu), although other species such as Pleioblastus amarus and Bambusa emeiensis have also been used medicinally. Ancient records predominantly noted its origin in Yizhou(present-day Chengdu and surrounding areas in Sichuan) and the Wuling region(between present-day Hunan, Guangdong, Guangxi and Jiangxi provinces), while contemporary sources are mainly from regions south of the Yangtze River and southwestern China. Traditionally, Bambusae Succus was harvested from bamboo that had grown for exactly one year, today, it can be collected year-round without strict age requirements. Ancient preparation methods included direct fire roasting or dry distillation, whereas modern industrial production employs dry distillation, reflux extraction, and percolation. In terms of quality evaluation, ancient texts considered a sweet taste to be superior, while today, clarity and transparency are prioritized. Historically, Bambusae Succus was characterized as sweet and cold nature, targeting the lung and stomach meridians, with uses evolving from clearing heat and resolving phlegm to nourishing Yin, moistening dryness, and relaxing tendons and unblocking meridians. Modern descriptions classify it as sweet, bitter, and cold in nature, affecting the heart, liver, and lung meridians, with functions including clearing heat, resolving phlegm, and facilitating orifices. It is indicated for conditions such as stroke with phlegm confusion, lung heat with phlegm congestion, convulsions, epilepsy, excessive phlegm in febrile diseases, high fever with thirst, irritability during pregnancy, and tetanus, with more clearly defined applications. Based on the results of the research, it is recommended that when developing and utilizing famous classical formulas containing Bambusae Succus, the one-year-old Phyllostachys nigra var. Henonis, which has been highly praised throughout history, should be selected as the source material. Industrial production should adopt the dry distillation method. Furthermore, in-depth research should be conducted on the modern technological characterization of the traditional quality control indicator of sweet taste, and reasonable modern quality control standards should be established.
8.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
ObjectiveTo optimize the extraction method for polysaccharides from turnip(Brassica rapa), and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction(BP-1) and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index, a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio, extraction time and extraction temperature, in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry(UV), gas chromatography-mass spectrometry(GC-MS), Congo red assay, and Fourier-transform infrared spectroscopy(FT-IR) to determine purity, monosaccharide composition, triple-helix structure, and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy(SEM) and atomic force microscopy(AFM). Zebrafish were divided into the blank group(adding E3 medium), and BP-1-1, BP-1-10, BP-1-50, BP-1-200, BP-1-1 000 groups(adding BP-1 solutions at concentrations of 1, 10, 50, 200, 1 000 mg·L-1, respectively), and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype, survival rate, malformation rate, and heart rate. Experimental animals were randomly divided into the blank group, model group, BP-1-10 group(10 mg·L-1), BP-1-50 group(50 mg·L-1), and BP-1-200 group(200 mg·L-1). The blank group was cultured in E3 medium, the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization(dpf) zebrafish embryos using 10 g·L-1 of glucose combined with 500 µmol·L-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution, while the blank and model groups received an equal volume of saline. Glucose and insulin(INS) levels were measured using enzyme-linked immunosorbent assay(ELISA) kits, the effects on the liver were observed by hematoxylin-eosin(HE) histopathological sections. The mRNA expression levels of glucagon(Glucagon), insulin(Insa), and phosphoenolpyruvate carboxykinase 1(PCK1) were detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). ResultsThe optimized extraction conditions were determined as follows:solid-to-liquid ratio of 1∶40(g·mL-1), extraction time of 66 min, and extraction temperature of 79 ℃. Under these conditions, the yield of turnip polysaccharides was (10.34±0.96)%. UV analysis indicated that BP-1 contained no proteins or nucleic acids, GC-MS analysis revealed that BP-1 consisted of six monosaccharides(arabinose, rhamnose, ribose, mannose, galactose and glucose). Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure, FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids, SEM analysis revealed an irregular flaky structure with a flat and smooth surface, AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L-1. Pharmacodynamic results showed that, compared with the blank group, the model group exhibited significantly increased glucose levels and significantly decreased INS levels(P<0.01). Compared with the model group, the BP-1-50 group significantly reduced glucose levels and increased INS levels(P<0.05). Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal, with hepatocytes appearing plump. Real-time PCR results showed that, compared with the blank group, the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1, and significantly downregulated mRNA expression of Insa(P<0.01). Compared with the model group, the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1, and significantly upregulated mRNA expression of Insa(P<0.01). ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate, is simple to operate, has low costs, making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides, contains α-glycosidic bonds and uronic acids, exhibits hypoglycemic activity, and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.
9.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
ObjectiveTo optimize the extraction method for polysaccharides from turnip(Brassica rapa), and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction(BP-1) and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index, a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio, extraction time and extraction temperature, in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry(UV), gas chromatography-mass spectrometry(GC-MS), Congo red assay, and Fourier-transform infrared spectroscopy(FT-IR) to determine purity, monosaccharide composition, triple-helix structure, and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy(SEM) and atomic force microscopy(AFM). Zebrafish were divided into the blank group(adding E3 medium), and BP-1-1, BP-1-10, BP-1-50, BP-1-200, BP-1-1 000 groups(adding BP-1 solutions at concentrations of 1, 10, 50, 200, 1 000 mg·L-1, respectively), and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype, survival rate, malformation rate, and heart rate. Experimental animals were randomly divided into the blank group, model group, BP-1-10 group(10 mg·L-1), BP-1-50 group(50 mg·L-1), and BP-1-200 group(200 mg·L-1). The blank group was cultured in E3 medium, the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization(dpf) zebrafish embryos using 10 g·L-1 of glucose combined with 500 µmol·L-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution, while the blank and model groups received an equal volume of saline. Glucose and insulin(INS) levels were measured using enzyme-linked immunosorbent assay(ELISA) kits, the effects on the liver were observed by hematoxylin-eosin(HE) histopathological sections. The mRNA expression levels of glucagon(Glucagon), insulin(Insa), and phosphoenolpyruvate carboxykinase 1(PCK1) were detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). ResultsThe optimized extraction conditions were determined as follows:solid-to-liquid ratio of 1∶40(g·mL-1), extraction time of 66 min, and extraction temperature of 79 ℃. Under these conditions, the yield of turnip polysaccharides was (10.34±0.96)%. UV analysis indicated that BP-1 contained no proteins or nucleic acids, GC-MS analysis revealed that BP-1 consisted of six monosaccharides(arabinose, rhamnose, ribose, mannose, galactose and glucose). Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure, FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids, SEM analysis revealed an irregular flaky structure with a flat and smooth surface, AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L-1. Pharmacodynamic results showed that, compared with the blank group, the model group exhibited significantly increased glucose levels and significantly decreased INS levels(P<0.01). Compared with the model group, the BP-1-50 group significantly reduced glucose levels and increased INS levels(P<0.05). Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal, with hepatocytes appearing plump. Real-time PCR results showed that, compared with the blank group, the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1, and significantly downregulated mRNA expression of Insa(P<0.01). Compared with the model group, the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1, and significantly upregulated mRNA expression of Insa(P<0.01). ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate, is simple to operate, has low costs, making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides, contains α-glycosidic bonds and uronic acids, exhibits hypoglycemic activity, and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.
10.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.


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