1.Effect of Danggui Buxuetang on PINK1/Parkin Signaling Pathway of Vascular Dementia Rats
Guifang QI ; Yue JIANG ; Yunxiang TAN ; Nanbu WANG ; Xinghua CHEN ; Ting WAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):15-24
ObjectiveTo investigate the potential mechanism of Danggui Buxuetang (DBT) in the treatment of vascular dementia (VAD). MethodsSixty male SD rats were randomly assigned to the sham-operated group, model group, DBT low-, medium-, and high-dose groups, and the donepezil group. Except for the sham-operated group, rats in all other groups underwent bilateral common carotid artery ligation. After successful modeling, DBT was administered at doses of 9.2, 18.4, 36.8 g·kg-1 for the low-, medium-, and high-dose groups, respectively, while the donepezil group received 3 mg·kg-1 donepezil solution by gavage once daily. After 4 consecutive weeks of drug treatment, rats underwent the Morris water maze test, novel object recognition test, Nissl staining to observe hippocampal neurons, and immunofluorescence staining to detect the expression of neuronal nuclear protein (NeuN) in the hippocampus. Western blot was used to assess the expression of PTEN-induced kinase 1 (PINK1), Parkin, microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax). Transmission electron microscopy was used to observe hippocampal neuronal ultrastructure. Real-time PCR was used to detect the expression of NADPH oxidase subunits p22phox and p47phox in hippocampal tissues. The levels of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), and total antioxidant capacity were measured to evaluate oxidative stress levels. ResultsIn the Morris water maze test, escape latency changed significantly over time in all groups except the model group. Compared with the sham-operated group, the model group showed significantly prolonged escape latency (P<0.01). Compared with the model group, rats in the DBT groups and the donepezil group exhibited significantly shorter escape latency (P<0.05, P<0.01). The number of crossings over the original platform was significantly reduced in the model group compared with the sham-operated group (P<0.01), whereas rats in the DBT and donepezil groups showed significantly increased platform crossings compared with the model group (P<0.05, P<0.01). Compared with the sham-operated group, exploration time of new objects was significantly reduced in the model group (P<0.01). Compared with the model group, exploration time of new objects increased significantly in the medium- and high-dose DBT groups and the donepezil group (P<0.05, P<0.01), while no significant change was observed in the low-dose DBT group. Compared with the high-dose DBT group, rats in the donepezil group had significantly prolonged escape latency and reduced platform crossings and new-object exploration time (P<0.05). Nissl staining showed decreased density of healthy neurons in the CA1 and CA3 regions of the hippocampus in the model group, with loss of Nissl bodies and nuclear atrophy or disappearance. In the high-dose DBT group, neuronal density in CA1 and CA3 increased, with neurons arranged closely and displaying normal morphology. Immunofluorescence showed that compared with the sham-operated group, the hippocampal NeuN⁺ cell count in the VAD model group was significantly decreased(P<0.01), compared with the VAD model group, the hippocampal NeuN⁺ cell count in the high-dose DBT group was significantly increased(P<0.01). Compared with the sham-operated group, the expression of PINK1, Parkin, LC3Ⅱ, and Bax proteins was significantly increased(P<0.01), while the expression of Bcl-2 was significantly decreased in the VAD model group(P<0.01). Compared with the VAD model group, the high-dose DBT group showed significantly decreased expression of PINK1, Parkin, LC3Ⅱ, and Bax proteins(P<0.01)and significantly upregulated Bcl-2 expression(P<0.01). The medium-dose DBT group exhibited significantly reduced expression of Parkin, LC3Ⅱ, and Bax proteins(P<0.05,P<0.01) and significantly increased Bcl-2 expression(P<0.01), while no statistically significant differences were observed in the low-dose DBT group. Transmission electron microscopy showed mitochondrial pyknosis, thickened cristae, increased electron density, and the presence of mitochondrial autophagy in the model group. In contrast, hippocampal neurons in the high-dose DBT group contained abundant mitochondria with intact morphology, clear cristae, and uniform matrix. Compared with the sham-operated group, total antioxidant capacity, SOD activity, and GSH levels were significantly decreased, while MDA levels were significantly increased in the model group (P<0.01). Compared with the model group, total antioxidant capacity and antioxidant levels (SOD, GSH) increased significantly, and MDA decreased significantly in the medium- and high-dose DBT groups (P<0.01), while no significant changes were observed in the low-dose DBT group. Compared with the sham-operated group, mRNA expression of p22phox and p47phox was significantly increased in the model group (P<0.01). Compared with the model group, expression of p22phox and p47phox was significantly decreased in the DBT groups (P<0.05, P<0.01). ConclusionDBT may exert neuroprotective effects by regulating PINK1/Parkin-mediated mitochondrial autophagy, thereby improving learning and memory abilities and treating VAD.
2.Mechanisms of Sini San in Regulation of Gut Microbiota Against Depression and Liver Injury in CUMS Rats
Junling LI ; Yan ZHANG ; Lei WANG ; Fang QI ; Zhenzhen CHEN ; Tianxing CHEN ; Yuhang LIU ; Xueying WANG ; Xianwen TANG ; Yubo LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):33-40
ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley (SD) rats were randomly divided into a normal group, model group (M), Sini San group (MS, 2.5 g·kg-1), and fluoxetine group (MF, 2 mg·kg-1). Except for the normal group, rats in the other three groups were subjected to chronic unpredictable mild stress (CUMS). After 8 weeks, the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum corticosterone (CORT), adrenocorticotropic hormone (ACTH), corticotropin-releasing factor (CRF), lipopolysaccharide (LPS), Zonulin, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), γ-aminobutyric acid (GABA) levels in the hippocampus and prefrontal cortex, and brain-derived neurotrophic factor (BDNF) levels in the hippocampus. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy, and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group, the sucrose preference of rats in the model group was significantly reduced (P0.01), whereas it was significantly increased in the Sini San group compared with the model group (P0.05). Compared with the normal group, hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased (P0.05), and compared with the model group, both were significantly increased in the Sini San group (P0.05, P0.01). Compared with the normal group, serum LPS and Zonulin levels in the model group were significantly increased (P0.05, P0.01), and compared with the model group, Zonulin levels in the Sini San group were significantly decreased (P0.05). No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group, widened and blurred tight junctions, sparse and shortened microvilli, and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group, which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators, liver function, and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.
3.Mechanisms of Sini San in Regulation of Gut Microbiota Against Depression and Liver Injury in CUMS Rats
Junling LI ; Yan ZHANG ; Lei WANG ; Fang QI ; Zhenzhen CHEN ; Tianxing CHEN ; Yuhang LIU ; Xueying WANG ; Xianwen TANG ; Yubo LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):33-40
ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley (SD) rats were randomly divided into a normal group, model group (M), Sini San group (MS, 2.5 g·kg-1), and fluoxetine group (MF, 2 mg·kg-1). Except for the normal group, rats in the other three groups were subjected to chronic unpredictable mild stress (CUMS). After 8 weeks, the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum corticosterone (CORT), adrenocorticotropic hormone (ACTH), corticotropin-releasing factor (CRF), lipopolysaccharide (LPS), Zonulin, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), γ-aminobutyric acid (GABA) levels in the hippocampus and prefrontal cortex, and brain-derived neurotrophic factor (BDNF) levels in the hippocampus. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy, and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group, the sucrose preference of rats in the model group was significantly reduced (P<0.01), whereas it was significantly increased in the Sini San group compared with the model group (P<0.05). Compared with the normal group, hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased (P<0.05), and compared with the model group, both were significantly increased in the Sini San group (P<0.05, P<0.01). Compared with the normal group, serum LPS and Zonulin levels in the model group were significantly increased (P<0.05, P<0.01), and compared with the model group, Zonulin levels in the Sini San group were significantly decreased (P<0.05). No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group, widened and blurred tight junctions, sparse and shortened microvilli, and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group, which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators, liver function, and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.
4.Effect and Mechanism of Schisandrae Chinensis Fructus Lignans on Behavior of Schizophrenic Mice
Jiaqi LI ; Xi CHEN ; Siwei WANG ; Qi WANG ; Yiting LIU ; Ziyan GUO ; Zilong LUN ; Chengyi ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):65-71
ObjectiveTo investigate the effects of Schisandrae Chinensis Fructus lignans on schizophrenia induced by dizocilpine maleate (MK-801) in mice and to clarify its mechanism. MethodsMale mice of 4-6 weeks old were randomized into blank, model, positive drug, and low-, medium-, and high-dose (40, 80, 160 mg·kg-1, respectively) Schisandrae Chinensis Fructus lignans groups. The blank group was administrated with distilled water, and the other groups were injected with 0.5 mg·kg-1 MK-801 to induce schizophrenia symptoms. Meanwhile, risperidone was injected at 0.2 mg·kg-1 in the positive drug group, and mice in the intervention groups were injected with corresponding drugs for 14 consecutive days. The behavioral changes of mice were observed by autonomous activity test, open field test, forced swimming test, and water maze test. The levels of dopamine (DA) and 5-hydroxytryptamine (5-HT) in the brain and tumor necrosis factor-α (TNF-α) and nuclear factor-κB (NF-κB) in peripheral blood were quantified by enzyme-linked immunosorbent assay (ELISA). The changes in the prefrontal lobe of mice were observed by hematoxylin-eosin staining, and the changes of the hippocampal tissue were observed by Nissl staining. The protein levels of silencing information regulatory factor 1 (SIRT1) and forkhead box protein O3a (FoxO3a) in the hippocampus of mice were determined by Western blot. ResultsCompared with the model group, low, medium, and high doses of Schisandrae Chinensis Fructus lignans reduced the total number of autonomous activities, total distance in the open field test, immobile time in the forced swimming test, and levels of TNF-α and NF-κB in peripheral blood (P<0.05), while increasing the number of platform crossings in the water maze test and DA and 5-HT levels in the brain tissue (P<0.05). Compared with the model group, risperidone and low, medium, and high doses of Schisandrae Chinensis Fructus lignans improve the neural cell morphology in the CA1 region, with full cells in neatly dense arrangement and exhibiting clear membrane boundary. Schisandrae Chinensis Fructus lignans inhibited the expression of SIRT 1 and FoxO3a in the hippocampus (P<0.05). ConclusionTo sum up, Schisandrae Chinensis Fructus lignans may improve the behavior of schizophrenic mice by activating the SIRT1/FoxO3a signaling pathway to exert neuroprotective effects.
5.Comparison of Wild and Cultivated Gardeniae Fructus Based on Traditional Quality Evaluation
Yuanjun SHANG ; Bo GENG ; Xin CHEN ; Qi WANG ; Guohua ZHENG ; Chun LI ; Zhilai ZHAN ; Junjie HU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):225-234
ObjectiveBased on traditional quality evaluation of Gardeniae Fructus(GF) recorded in historical materia medica, this study systematically compared the quality differences between wild and cultivated GF from morphological characteristics, microscopic features, and contents of primary and secondary metabolites. MethodsVernier calipers and analytical balances were used to measure the length, diameter and individual fruit weight of wild and cultivated GF, and the aspect ratio was calculated. A colorimeter was used to determine the chromaticity value of wild and cultivated GF, and the paraffin sections of them were prepared by safranin-fast green staining and examined under an optical microscope to observe their microstructure. Subsequently, the contents of water-soluble and alcohol-soluble extracts of wild and cultivated GF were detected by hot immersion method under the general rule 2201 in volume Ⅳ of the 2020 edition of the Pharmacopoeia of the People's Republic of China, the starch content was measured by anthrone colorimetric method, the content of total polysaccharides was determined by phenol-sulfuric acid colorimetric method, the sucrose content was determined by high performance liquid chromatography coupled with evaporative light scattering detection(HPLC-ELSD), and the contents of representative components in them were measured by ultra-performance liquid chromatography(UPLC). Finally, correlation analysis was conducted between quality traits and phenotypic traits, combined with multivariate statistical analysis methods such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), key differential components between wild and cultivated GF were screened. ResultsIn terms of traits, the wild GF fruits were smaller, exhibiting reddish yellow or brownish red hues with significant variation between batches. While the cultivated GF fruits are larger, displaying deeper orange-red or brownish red. The diameter and individual fruit weight of cultivated GF were significantly greater than those of wild GF, while the blue-yellow value(b*) of wild GF was significantly higher than that of cultivated GF. In the microstructure, the mesocarp of wild GF contained numerous scattered calcium oxalate cluster crystals, while the endocarp contained stone cell class round, polygonal or tangential prolongation, undeveloped seeds were visible within the fruit. In contrast, the mesocarp of cultivated GF contained few calcium oxalate cluster crystals, or some batches exhibited extremely numerous cluster crystals. The stone cells in the endocarp were predominantly round-like, with the innermost layer arranged in a grid pattern. Seeds were basically mature, and only a few immature seeds existed in some batches. Regarding primary metabolite content, wild GF exhibited significantly higher total polysaccharide level than cultivated GF(P<0.01). In category-specific component content, wild GF exhibited significantly higher levels of total flavonoids and total polyphenols compared to cultivated GF(P<0.01). Analysis of 12 secondary metabolites revealed that wild GF exhibited significantly higher levels of Shanzhiside, deacetyl asperulosidic acid methyl ester, gardenoside and chlorogenic acid compared to cultivated GF(P<0.01). Conversely, the contents of genipin 1-gentiobioside, geniposide and genipin were significantly lower in wild GF(P<0.01). ConclusionThere are significant differences between wild and cultivated GF in terms of traits, microstructure, and contents of primary and secondary metabolites. At present, the quality evaluation system of cultivated GF remains incomplete, and this study provides a reference for guiding the production of high-quality GF medicinal materials.
6.Clinical Advantages of Traditional Chinese Medicine in Treatment of Childhood Simple Obesity: Insights from Expert Consensus
Qi ZHANG ; Yingke LIU ; Xiaoxiao ZHANG ; Guichen NI ; Heyin XIAO ; Junhong WANG ; Liqun WU ; Zhanfeng YAN ; Kundi WANG ; Jiajia CHEN ; Hong ZHENG ; Xinying GAO ; Liya WEI ; Qiang HE ; Qian ZHAO ; Huimin SU ; Zhaolan LIU ; Dafeng LONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):238-245
Childhood simple obesity has become a significant public health issue in China. Modern medicine primarily relies on lifestyle interventions and often suffers from poor long-term compliance, while pharmacological options are limited and associated with potential adverse effects. Traditional Chinese Medicine (TCM) has a long history in the prevention and management of this condition, demonstrating eight distinct advantages, including systematic theoretical foundation, diversified therapeutic approaches, definite therapeutic efficacy, high safety profile, good patient compliance, comprehensive intervention strategies, emphasis on prevention, and stepwise treatment protocols. Additionally, TCM is characterized by six distinctive features: the use of natural medicinal substances, non-invasive external therapies, integration of medicinal dietetics, simple exercise regimens, precise syndrome differentiation, and diverse dosage forms. By combining internal and external treatments, TCM facilitates individualized regimen adjustment and holistic regulation, demonstrating remarkable effects in improving obesity-related metabolic indicators, regulating constitutional imbalance, and promoting healthy behaviors. However, challenges remain, such as inconsistent operational standards, insufficient high-quality clinical evidence, and a gap between basic research and clinical application. Future efforts should focus on accelerating the standardization of TCM diagnosis and treatment, conducting multicenter randomized controlled trials, and fostering interdisciplinary integration, so as to enhance the scientific validity and international recognition of TCM in the prevention and treatment of childhood obesity.
7.Reporting Status of Clinical Practice Guideline Protocols: A Systematic Analysis
Huayu ZHANG ; Xufei LUO ; Hui LIU ; Qi ZHOU ; Yishan QIN ; Ye WANG ; Yuanyuan YAO ; Haodong LI ; Xiaohui WANG ; Yaolong CHEN
Medical Journal of Peking Union Medical College Hospital 2026;17(1):255-262
To systematically analyzed the reporting status of core elements in publicly available clinical practice guideline(hereafter referred to as "guideline") protocols published domestically and internationally over the past decade, identified existing problems, and provided evidence to inform the standardized writing and publication of future guideline protocols. A systematic search was conducted in Chinese and English databases for clinical practice guideline protocols published during the past ten years. The basic characteristics and reporting of core elements—including registration information, conflict of interest management, evidence grading, development process and timeline planning, as well as dissemination and implementation—were extracted and analyzed. Chi-square tests were performed to explore associations between protocol characteristics and the reporting of core elements. A total of 94 guideline protocols were included, of which 67 were in Chinese(71.28%) and 27 were in English(28.72%). Overall, 82.98% of the guideline protocols were registered, 92.55% reported management of conflicts of interest, 97.87% reported evidence searching, 88.30% reported evidence grading, and 89.36% described dissemination and implementation strategies. However, only 55.32% reported the guideline development process, and merely 23.40% reported timeline planning. Further analysis indicated that the reporting of registration, evidence searching, development process, and timeline planning was associated with year of publication. Differences were observed between domestic and international guidelines in reporting registration, conflict of interest management, development process, time planning, and dissemination and implementation. Guidelines intended for development exhibited higher reporting rates for registration, development process, and dissemination and implementation compared to those planned for updating or adaptation. Although current guideline protocols demonstrate relatively adequate reporting of methodological elements, deficiencies remain in development process and timeline planning. Future efforts should focus on promoting the publication and standardized reporting of guideline protocols, enhancing the international recognition of registration platforms, and strengthening the development process and timeline planning to advance the scientific rigor and transparency of guideline development.
8.Mechanism of drug-containing serum of Dianxianqing granules in inhibiting microglial ferroptosis
Guangkun FAN ; Yue QI ; Jixian WANG ; Wei CHEN ; Chunpeng XIA ; Yihang WANG ; Yue ZHAO ; Yang AN
China Pharmacy 2026;37(3):317-323
OBJECTIVE To explore the potential mechanism by which drug-containing serum of Dianxianqing granules (DXQ) inhibits microglial ferroptosis. METHODS Male SD rats were given normal saline and Dianxianqing granules solution via intragastric administration to prepare normal serum and DXQ, respectively. Mice microglia BV2 cells were collected and successfully transfected with a negative control small interfering RNA (si-NC), and then they were included in the si-NC group and cultured under normal conditions. Cells successfully transfected with small interfering RNA targeting glutathione peroxidase 4 (GPX4) (si-GPX4) were divided into the si-GPX4 group, the CsA group (treated with 1 μmol/L cyclosporine A), and the DXQ- L, DXQ-M and DXQ-H groups (treated with 5%, 7% and 10% DXQ, respectively). These groups were subsequently treated with their corresponding drug solutions and ferroptosis inducer Erastin (10 μmol/L). The intracellular levels of total iron ions, glutathione (GSH), reactive oxygen species (ROS), and the expression of mitochondrial superoxide were determined in each group after 48 h of treatment. Additionally, mitochondrial membrane potential, the opening degree of mitochondrial permeability transition pore (MPTP), and mRNA expressions of GPX4 and cyclophilin D (CypD) were detected. Furthermore, the expressions of ferroptosis-related proteins[GPX4, transferrin receptor 1 (TfR1) and ferritin heavy chain 1 (FTH1)], as well as MPTP-related proteins [adenine nucleotide translocator (ANT), cytochrome C (CytC), mitochondrial calcium uniporter (MCU) and CypD] were assessed. RESULTS Compared with si-NC group, the levels of total iron ions and ROS, the expression level of mitochondrial superoxide, the opening degree of MPTP, protein and its mRNA expressions of CypD as well as protein expressions of TfR1 and MCU were increased or up-regulated significantly (P<0.01); however, GSH content, mitochondrial membrane potential, protein and mRNA expressions of GPX4, and protein expressions of FTH1, ANT and CytC were decreased or down-regulated significantly (P<0.01). Compared with the si-GPX4 group, the cells in the DXQ-M, DXQ-H groups showed a general improvement in the above quantitative indicators (P<0.01 or P<0.05). CONCLUSIONS DXQ can enhance antioxidant capacity by activating the GSH/GPX4 pathway, regulate the expressions of TfR1 and FTH1 protein to correct iron ion homeostasis, inhibit excessive opening of MPTP to improve mitochondrial function, and ultimately suppress microglial ferroptosis.
9.Research advances on the intergenerational transmission of adolescent health behaviors
WANG Yating, CAO Meijuan, ZENG Yaling, CHEN Qi
Chinese Journal of School Health 2026;47(2):291-295
Abstract
To improve adolescent health behavior, the study summarizes and analyzes the performance, pathways of transmission, and influencing factors of the intergenerational transmission of adolescent health behaviors from the perspective of intergenerational transmission. The study emphasizes the need to deepen research on the intergenerational transmission of adolescent health behaviors, promote multidisciplinary and cross team collaboration, and shift adolescent health care from individual focused care to a holistic approach that prioritizes family and community culture. Simultaneously, an action framework should be established to block the intergenerational transmission of health risk behaviors, with a focus on childhood and adolescence. Additionally, parent-child participatory health education and health promotion activities should be carried out under a tripartite coordinated intervention model involving the community, school, and family, collectively fostering the development of healthy behaviors among adolescents.
10.Epigenetic mechanism of Diwu Yanggan Capsule in improving liver regeneration microenvironment in a rat model of liver cancer
Minggang WANG ; Jiamei DONG ; Zhihua YE ; Xiang GAO ; Qi CHEN ; Xiaoqiao YU ; Hanmin LI
Journal of Clinical Hepatology 2026;42(2):362-371
ObjectiveTo investigate the epigenetic mechanism of Diwu Yanggan Capsule in improving liver regeneration microenvironment in a rat model of liver cancer by regulating DNA methylation, and to provide a basis for scientific clinical medication. MethodsA total of 48 specific pathogen-free Sprague-Dawley rats were divided into normal group, model group, and Diwu Yanggan Capsule group using a random number table, with 16 rats in each group. The Solt-Farber two-step method was used to establish a rat model of liver cancer. The rats in the Diwu Yanggan Capsule group were given Diwu Yanggan Capsule at a dose of 750 mg/kg/d by gavage, and those in the normal group and the model group were given an equal volume of normal saline by gavage. Liver tissue samples were collected from each group of rats after 16 weeks of continuous intervention; DNA methylation chips were used to analyze the change in DNA methylation in liver tissue, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used for data analysis. In addition, the MeDIP-PCR technique was used to detect the changes in candidate differentially methylated genes such as YWHAB, ADCK2, ERLIN2, SEMA3B, and TPH2 in the liver tissue of rats, and Western blot and RT-qPCR were used to verify the expression of key methylated genes. The independent-samples t test was used for comparison of continuous data between two groups, and a one-way analysis of variance was used for comparison between multiple groups, while the least significant difference t-test was used for further comparison between two groups. ResultsThe DNA methylation chip analysis showed that compared with the normal group, the model group had significant methylation changes in the promoter region of 2 422 genes in liver tissue of rats. The GO functional enrichment analysis and the KEGG pathway enrichment analysis showed that these differentially methylated genes were significantly enriched in metabolic pathways such as steroid hormone biosynthesis and drug metabolism-cytochrome P450. Compared with the model group, the Diwu Yanggan Capsule group had significant reversal of promoter methylation in 1 650 genes, and the KEGG enrichment analysis showed that these genes were mainly involved in the pathways closely associated with cell proliferation, apoptosis, and microenvironment regulation, such as the calcium ion signaling pathway, the cAMP signaling pathway, and the extracellular factor signaling pathway. Compared with the model group, the Diwu Yanggan Capsule group had a significant increase in the promoter methylation level of the ADCK2 gene (P<0.05) and significant reductions in the promoter methylation levels of the ERLIN2 and TPH2 genes (all P<0.05). Compared with the model group, the Diwu Yanggan Capsule group had significant reductions in the mRNA expression levels and the protein expression levels of the ADCK2 (all P<0.05). ConclusionAbnormal DNA methylation in liver tissue participates in the development and progression of liver cancer. The effect of Diwu Yanggan Capsule on DNA methylation level is an important epigenetic mechanism for its effect in the prevention and treatment of liver cancer.


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