1.Huangqi Jianzhongtang Regulates Polarization of Macrophages M1/M2 and Improves Fat Consumption in Cancer Cachexia Mice
Zhiyan FANG ; Haiyan ZHU ; Wenying HUAI ; Cong HUANG ; Ruocong YANG ; Haiyan YU ; Tiane ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):61-69
ObjectiveTo investigate the effects of Huangqi Jianzhongtang (HQJZ) on macrophage polarization and fat consumption in cancer cachexia (CC) mice. MethodsUltra-performance liquid chromatography-quadrupole/electrostatic field Orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was used to control the quality of HQJZ. (1) In vitro experiment: HQJZ-containing serum was prepared, and the optimal concentration was determined by cytotoxicity assay. Mouse monocyte-derived macrophages (RAW264.7) were cultured and randomly divided into six groups, including a blank group, a classically activated macrophages (M1) group, an alternatively activated macrophages (M2) group, a HQJZ + blank group, a HQJZ+M1 group, and a HQJZ + M2 group. The relative expression of macrophage marker genes CD86, inducible nitric oxide synthase (iNOS), CD206, and arginase-1 (Arg1) was detected by real-time quantitative polymerase chain reaction (Real-time PCR ). (2) In vivo experiment: Thirty-two BALB/c mice were randomly divided into a control group, a model group, a medroxyprogesterone acetate (MPA) group, and a HQJZ group. Except for the control group, the other mice were injected with CT-26 colon cancer cells to establish a CC model. Mice in the MPA and HQJZ groups were given MPA (0.13 g·kg-1·d-1) or HQJZ (13.13 g·kg-1·d-1) by gavage, respectively, while mice in the control and model groups were given an equal volume of saline by gavage, with interventions continued for 10 d. Real-time PCR was used to detect the expression of macrophage markers (iNOS, Arg1, CD86, CD206) and fat browning-related genes uncoupling protein 1 (UCP1) and peroxisome proliferator-activated receptor γ (PPARγ) in epididymal adipose tissue. Western blot (WB) was used to detect protein expression levels of UCP1 and PPARγ. Micro-computed tomography (micro-CT) was used to measure residual fat volume, and hematoxylin-eosin (HE) staining was used to assess fat browning and calculate pathological scores. ResultsIn vitro, the dominant effective concentration of HQJZ-containing serum was 12.5%. Real-time PCR results showed that, compared with the blank group, Arg1 expression decreased in the HQJZ+blank group (P<0.05), CD206 showed a downward trend without statistical significance, while iNOS and CD86 expression were significantly increased (P<0.05). Compared with the M1 group, Arg1 and CD206 expression decreased in the HQJZ+M1 group (P<0.05). Compared with the M2 group, CD206 expression decreased in the HQJZ+M2 group (P<0.05), CD86 expression increased significantly (P<0.01). In vivo, Real-time PCR results showed that, compared with the control group, CD86 and CD206 expression levels were significantly increased in the model group (P<0.01). Compared with the model group, CD206 expression in the MPA group was significantly decreased (P<0.01). In the HQJZ group, CD206 was significantly decreased (P<0.01). WB results showed that, compared with the model group, protein expression of UCP1 and PPARγ was significantly reduced in the HQJZ group (P<0.05, P<0.01). micro-CT results showed that the total white fat volume in the HQJZ group was greater than that in the model group (P<0.05). HE staining results showed that pathological scores in the HQJZ group were lower than those in the model group (P<0.05). ConclusionHQJZ may inhibit white adipose tissue browning by promoting macrophage M1 polarization and suppressing M2 polarization, thereby delaying fat consumption in CC mice.
2.Molecular Crosstalk Mechanisms of Shoutai Wan and Juyuan Jian on Maternal-fetal Interface Subcellular Clusters in CBA/J×DBA/2 Recurrent Pregnancy Loss Model
Jingxin GAO ; Qiuping CHEN ; Xiaoyan ZHENG ; Pengfei ZENG ; Rui ZHOU ; Yancai TANG ; Qian ZENG ; Wenli GUO ; Jinzhu HUANG ; Weijun DING ; Linwen DENG ; Hang ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):70-87
ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss (RPL) by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus (Shoutai Wan, Juyuan Jian)-of traditional Chinese medicine (TCM) at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c (blank group) and DBA/2 (modeling group) mice separately. Pregnant mice in the modeling group were randomly grouped as follows: high/low-dose Shoutai Wan, high/low-dose Juyuan Jian, model (RPL), and positive control (dydrogesterone), with 10 mice in each group. Starting from the day after the detection of the vaginal plug, mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention, the following indicators were measured. ① Macroscopic evaluation: general conditions, uterine wet weight, embryo loss rate, four coagulation parameters [prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), and thrombin time (TT)], and peripheral blood estradiol (E2) and progesterone (Pg) levels. The decidua with embryos was stained with hematoxylin-eosin (HE) and evaluated by transmission electron microscopy (TEM). The expression of B-cell lymphoma-2 (Bcl-2), vascular endothelial growth factor (VEGF), angiotensin Ⅱ (AngⅡ), matrix metalloproteinase-2 (MMP-2), interleukin-6 (IL-6), leukemia inhibitory factor (LIF), CXC chemokine ligand 12 (CXCL12), and microtubule-associated protein 1 light chain 3 homolog (LC3)Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level: The decidua with embryos from the blank, model, high-dose Shoutai Wan, and high-dose Juyuan Jian groups (6 mice per group, with 3 single-cell samples per group, totaling 24 mice) were analyzed by the BD Rhapsody™ platform, and the whole-cell atlas was drawn by uniform manifold approximation and projection (UMAP) dimensionality reduction clustering combined with the single-cell mouse cell atlas (scMCA). The differentially expressed genes (DEGs) and cell interaction networks were analyzed via Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and CellChat, and the protein-protein interaction (PPI) map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells (natural killer cells, NK cells) from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group, the RPL group showed an increase in the embryo loss rate (P<0.01), down-regulated expression of Bcl-2, LIF, MMP-2, and Vegf in the decidua with embryos (P<0.05), up-regulated protein levels of CXCL-12, AngⅡ, and IL-6 (P<0.05), blocked angiogenesis, apoptosis-inflammation imbalance, and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance, and Shoutai pill showed superior performance in restoring the E2 level to the Pg level (P<0.05). ② Single-cell transcriptome analysis indicated that compared with the blank group, the RPL group showed differences in multiple key cell populations such as decidual cells, trophoblast cells, endothelial cells, erythroblasts, NK cells, and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group, high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer (upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes) and macrophages (upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes) through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins, cell adhesion, and programmed cell death, thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells (up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes) and macrophages (up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes), which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D, CDH5, GDF, and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 (STAT3) and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis (TWEAK) signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7, a decreased proportion of reparative dNK4, and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1, thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method (Shoutai Wan) has an advantage in regulating the phenotypes of unfolded protein, cell adhesion, and programmed cell death, and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method (Juyuan Jian) has an advantage in regulating epithelial-mesenchymal transition (EMT), angiogenesis, and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.
3.Mechanism of Shenfu Xiongze Prescription in Regulating Autophagy Level to Intervene in Myocardial Remodeling in Rats via AMPK/mTOR Signaling Pathway
Xueqing WANG ; Wei ZHONG ; Liangliang PAN ; Caihong LI ; Man HAN ; Xiaowei YANG ; Yuanwang YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):136-144
ObjectiveTo explore the mechanism by which the Shenfu Xiongze prescription regulates autophagy in rats with myocardial remodeling through the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. MethodsA rat model of myocardial remodeling induced by isoprenaline (ISO) was established. Rats were divided into the blank group,the model group,the low-,medium-, and high-dose groups of Shenfu Xiongze prescription,and the captopril group, 6 rats in each group. Except for the blank group,the rat model of myocardial remodeling was established in the other groups by intraperitoneal injection of 2.5 mg·kg-1 ISO for 3 consecutive weeks. At the same time of modeling, the low-,medium-, and high-dose groups of Shenfu Xiongze prescription were administered the corresponding doses of Shenfu Xiongze prescription solution (8.4,16.8,and 33.6 g·kg-1),and the captopril group was administered captopril solution (25 mg·kg-1). As for the blank group and the model group, the same volume of normal saline was given. The treatment was continued for 3 weeks. Echocardiography was used to observe the cardiac structure and function,and the heart weight index was detected. Masson staining and hematoxylin-eosin (HE) staining were used to observe the pathological morphology changes of myocardial tissue. The levels of interleukin-6 (IL-6) and B-type natriuretic peptide (BNP) in serum were detected by enzyme-linked immunosorbent assay (ELISA). The expression of type Ⅰ collagen (Collagen Ⅰ),type Ⅲ collagen (Collagen Ⅲ),and microtubule-associated protein 1 light chain 3 (LC3) proteins in myocardial tissue was determined by immunohistochemistry. Autophagy was observed by transmission electron microscopy. The mRNA expression of Collagen Ⅰ,Collagen Ⅲ,α-smooth muscle actin (α-SMA),LC3,yeast Atg6 homolog protein (Beclin-1),AMPK,and mTOR in myocardial tissue was detected by quantitative real-time polymerase chain reaction (real-time PCR). The protein expression of Collagen Ⅰ,α-SMA,transforming growth factor-β1 (TGF-β1),LC3,Beclin-1,p62, phosphorylation(p)-AMPK,p-mTOR,AMPK,and mTOR was detected by Western blot. ResultsCompared with the normal group,rats in the model group exhibited significantly decreased values of ejection fraction (EF) and left ventricular fractional shortening (FS) (P<0.01), significantly increased values of left ventricular end-diastolic diameter (LVIDd) and left ventricular end-systolic diameter (LVIDs) (P<0.01). Additionally, the model group also showed increased degrees of inflammatory infiltration and fibrosis of myocardial tissue, significantly elevated levels of serum IL-6 and BNP (P<0.01), significantly increased mRNA and protein levels of Collagen Ⅰ,Collagen Ⅲ,α-SMA,and mTOR (P<0.01),and markedly decreased mRNA and protein levels of LC3,Beclin-1,and AMPK (P<0.05,P<0.01). Compared with the model group, the low-,medium-, and high-dose groups of Shenfu Xiongze prescription presented significantly elevated EF and FS values (P<0.01) and lowered LVIDd and LVIDs (P<0.05). In these groups, the inflammation and fibrosis were alleviated significantly. They also exhibited decreased serum levels of IL-6 and BNP (P<0.01), significantly reduced protein expression of Collagen Ⅰ, α-SMA, TGF-β1, p62, and p-mTOR (P<0.01), significantly decreased mRNA expression of Collagen Ⅰ, Collagen Ⅲ, α-SMA, and mTOR (P<0.01), and significantly increased mRNA and protein levels of LC3, Beclin-1, and AMPK (P<0.05,P<0.01). ConclusionThe Shenfu Xiongze prescription can improve the myocardial remodeling induced by ISO in rats by regulating the autophagy level,enhance cardiac function,and reduce inflammatory and fibrotic levels. This effect may be achieved through the AMPK/mTOR signaling pathway.
4.Joint Relation Extraction of Famous Medical Cases with CasRel Model Combining Entity Mapping and Data Augmentation
Yuxin LI ; Xinghua XIANG ; Hang YANG ; Dasheng LIU ; Jiaheng WANG ; Zhiwei ZHAO ; Jiaxu HAN ; Mengjie WU ; Qianzi CHE ; Wei YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):218-225
ObjectiveTo address the challenges of unstructured classical Chinese expressions, nested entity relationships, and limited annotated data in famous traditional Chinese medicine(TCM) case records, this study proposes a joint relation extraction framework that integrates data augmentation and entity mapping, aiming to support the construction of TCM diagnostic knowledge graphs and clinical pattern mining. MethodsWe developed an annotation structure for entities and their relationships in TCM case texts and applied a data augmentation strategy by incorporating multiple ancient texts to expand the relation extraction dataset. A cascade binary tagging framework for relation triple extraction(CasRel) model for TCM semantics was designed, integrating a pre-trained bidirectional encoder representations from transformers(BERT) layer for classical TCM texts to enhance semantic representation, and using a head entity-relation-tail entity mapping mechanism to address entity nesting and relation overlapping issues. ResultsExperimental results showed that the CasRel model, combining data augmentation and entity mapping, outperformed the pipeline-based Bert-Radical-Lexicon(BRL)-bidirectional long short-term memory(BiLSTM)-Attention model. The overall precision, recall, and F1-score across 12 relation types reached 65.73%, 64.03%, and 64.87%, which represent improvements of 14.26%, 7.98%, and 11.21% compared to the BRL-BiLSTM-Attention model, respectively. Notably, the F1-score for tongue syndrome relations increased by 22.68%(69.32%), and the prescription-syndrome relations performed the best with the F1-score of 70.10%. ConclusionThe proposed framework significantly improves the semantic representation and complex dependencies in TCM texts, offering a reusable technical framework for structured mining of TCM case records. The constructed knowledge graph can support clinical syndrome differentiation, prescription optimization, and drug compatibility, providing a methodological reference for TCM artificial intelligence research.
5.Clinical Application and Pharmacological Effects of Guizhi Fulingwan in Treatment of Uterine Fibroids: A Review
Xiang ZHANG ; Xiaoli WEN ; Biting CHENG ; Hongning LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):299-307
Uterine fibroids are a common benign tumor of the female reproductive system, characterized by increased menstrual flow, lower abdominal pain, and prolonged menstrual periods. Modern medicine believes that the onset of this disease is related to genetic factors, environmental factors, hormone levels, et al, while the specific mechanism remains unclear, and the prevention and treatment of uterine fibroids has become a hot topic of concern for many experts and scholars in the medical field. At present, the treatment of uterine fibroids in clinical practice is mainly based on hormone drugs and uterine artery embolization, and severe cases require hysterectomy. However, the use of hormone drugs for treatment has serious side effects and is prone to recurrence after surgery. Since hysterectomy can cause severe harm to women, it is necessary to explore safer and more effective treatment methods. Traditional Chinese medicine (TCM) has rich clinical experience in the treatment of uterine fibroids, advocating for syndrome differentiation and treatment. The TCM methods that regulate Qi and blood and balance Yin and Yang have been commonly adopted, with significant efficacy and minimal side effects, being more conducive to the recovery. Guizhi Fulingwan are first recorded in the Synopsis of the Golden Chamber written by the famous medical expert ZHANG Zhongjing during the Eastern Han dynasty. This prescription has the effects of activating blood, resolving stasis, and eliminating mass, and it is thus mainly used for treating abdominal mass in women. In recent years, Guizhi Fulingwan has also been applied in the clinical treatment of tumors and has demonstrated definite efficacy in the treatment of uterine fibroids. Studies have shown that the therapeutic mechanisms of Guizhi Fulingwan for uterine fibroids involve regulating cell proliferation and apoptosis, improving immune function, reducing inflammation, improving hemorheological indicators, inhibiting tumor angiogenesis, and regulating sex hormone levels. This article mainly reviews the treatment of uterine fibroids with Guizhi Fulingwan from three aspects: theoretical basis, clinical application, and pharmacological mechanism. It is expected to provide scientific research ideas and guidance for exploring the clinical treatment of uterine fibroids.
6.Ameliorative Effect of Wendantang Combined with Danshenyin and Dushentang on Ischemic Heart Disease with Phlegm-stasis Syndrome in Mice Based on Circulating Monocytes
Fenghe YANG ; Ziqi TIAN ; Zhiqian SONG ; Shitao PENG ; Wenjie LU ; Tao LIN ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):22-32
ObjectiveTo investigate the ameliorative effect of Wendantang combined with Danshenyin and Dushentang (WDD) on mice with ischemic heart disease (IHD) presenting phlegm-stasis syndrome based on the inflammatory phenotype and differentiation of circulating monocytes. MethodsA model of IHD with phlegm-stasis syndrome was established using left anterior descending coronary artery ligation supplemented with a high-fat diet. Eighty model mice were randomly assigned to the model group, WDD low-dose group (WDD-L), WDD medium-dose group (WDD-M), WDD high-dose group (WDD-H), and atorvastatin calcium tablet group, with 16 mice in each group. An additional 16 C57BL/6J mice were designated as the sham-operation group. The WDD groups received intragastric administration at doses of 8.91, 17.81, 35.62 g·kg-1, and the atorvastatin calcium tablet group received the corresponding drug at 1.3 mg·kg-1, twice daily. The sham-operation and model groups were given the same volume of pure water by gavage each day. After 5 consecutive weeks of administration, the cardiac index was calculated. Cardiac function was assessed by echocardiography. Myocardial histopathology was examined by hematoxylin-eosin (HE) staining. Serum N-terminal pro-B-type natriuretic peptide (pro-BNP) content was measured by enzyme-linked immunosorbent assay (ELISA). Hemorheological parameters were analyzed using an automated hemorheology analyzer. Serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were determined using an automated biochemical analyzer. Changes in circulating monocytes were detected by flow cytometry. Mouse bone marrow mononuclear cells were isolated in vitro and divided into blank group, model serum group, WDD-L drug-containing serum group, WDD-M drug-containing serum group, and WDD-H drug-containing serum group. CD36 expression and macrophage differentiation in each group were assessed by flow cytometry. The mechanism by which WDD mediates circulating monocyte differentiation was further explored using CD36 knockdown/overexpression RAW264.7 cell lines. ResultsCompared with the sham-operation group, the model group showed a significantly increased cardiac index (P0.01), significantly decreased fractional shortening (FS) (P0.01), and significantly increased left ventricular end-diastolic internal diameter (LVDD) and left ventricular end-systolic internal diameter (LVDS) (P0.01). Cardiomyocytes exhibited marked deformation and necrosis with inflammatory cell infiltration. Serum pro-BNP levels were significantly elevated (P0.01), and whole-blood viscosity (BV) at high, medium, and low shear rates was significantly increased (P0.01). Compared with the model group, the WDD groups showed significantly reduced cardiac index (P0.05, P0.01), significantly increased FS (P0.05, P0.01), significantly decreased LVDD and LVDS (P0.01), markedly improved cardiomyocyte morphology, significantly reduced inflammatory infiltration, significantly decreased serum pro-BNP levels (P0.01), and significantly decreased BV at high, medium, and low shear rates (P0.01), with the most pronounced improvement observed in the WDD-M group. Compared with the sham-operation group, TC, TG, and LDL levels were significantly increased in the model group (P0.05, P0.01), while HDL levels were significantly decreased (P0.05). After WDD-H treatment, TC, TG, and LDL levels were significantly reduced and HDL levels were significantly increased in mice (P0.05, P0.01). Compared with the sham-operation group, classical monocytes in blood and bone marrow and intermediate monocytes in blood were significantly increased in the model group (P0.01), whereas intermediate monocytes in bone marrow and non-classical monocytes in blood were significantly decreased (P0.01). After WDD administration, all circulating monocyte subsets in blood and bone marrow were significantly alleviated (P0.05, P0.01), with the WDD-M group showing the optimal effect. In vitro, compared with the blank group, CD36 expression on bone marrow monocytes and the proportion of differentiated macrophages were significantly increased in the model serum group (P0.01), and CD36 expression was significantly upregulated on RAW264.7 cells (P0.01). Compared with the model serum group, all drug-containing serum groups exhibited significantly reduced CD36 expression on bone marrow monocytes and significantly reduced macrophage differentiation (P0.01). WDD downregulated CD36 expression in both CD36 knockdown and overexpression RAW264.7 cell lines (P0.05, P0.01), with the strongest regulatory effect observed in the WDD-M drug-containing serum group. ConclusionWDD can significantly improve the manifestations of phlegm-stasis syndrome in IHD mice and reduce the proportion of classical circulating monocytes. Its mechanism may be related to the inhibition of CD36 expression on classical circulating monocytes.
7.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.
8.Clinical Efficacy of Qi-regulating and Phlegm-removing Method(Liu Junzitang Combined with Linggang Wuwei Jiangxintang) in Treating AECOPD with Increased EOS
Renjie HUANG ; Wangqin YU ; Wuyinuo TANG ; Hong SONG ; Lyuyuan HE ; Wenbo LIN ; Guanyi WU ; Hang HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):149-156
ObjectiveTo assess the efficacy and safety of the Qi-regulating and phlegm-removing method(Liu Junzitang Combined with Linggang Wuwei Jiangxintang) for treating acute exacerbations of chronic obstructive pulmonary disease (AECOPD) with increased eosinophils (EOS). MethodsSixty-eight AECOPD patients with increased EOS who were hospitalized in the Department of Pulmonary Diseases of Jinhua Traditional Chinese Medicine Hospital from April 2023 to April 2024 were recruited and randomly assigned to an experimental group (EG) or a control group (CG). Both groups received conventional Western medicine, with the EG additionally receiving Liujunzitang and Linggan Wuwei Jiangxintang. The therapeutic efficacy indicators were measured after the treatment. The main therapeutic efficacy indicators included partial pressure of oxygen (PaO2) and partial pressure of carbon dioxide (PaCO2). The secondary efficacy indicators included the TCM symptom scores, the COPD Assessment Test (CAT) score, the Modified Medical Research Council (mMRC) Dyspnea Scale score, and the length of hospital stay. The indicators were measured at baseline and on days 3 and 7 of intervention. The safety was evaluated based on the adverse events. ResultsBaseline characteristics were not statistically different between the two groups. Compared with CG, EG showed no significant difference in PaO2 (P=0.773), PaCO2 (P=0.632) and or CAT score (P=0.336) at on day 3 but better PaO2 (P=0.004), PaCO2 (P=0.008), and CAT score (P=0.013) were significantly better at on day 7. Compared with CGAfter treatment, EG had lower TCM syndrome scores of than CG EG on day 3 (P=0.005) and day 7 were significantly decreased (P0.001). There was no significant difference in mMRC score between the two groups on day 3 (P=0.514) and day 7 (P=0.176) as wasor the length of hospital stay (P=0.915). The generalized linear mixed model (GLMM) showed that compared with CG, EG had significant improvements over time in PaO2, PaCO2, TCM syndrome symptom scores, CAT score, and mMRC score. ConclusionRegulating qi Qi and removing phlegm combined with conventional Western medicine can significantly alleviateimprove the clinical symptoms and improve the lung function of AECOPD patients with increased EOS increased AECOPDwhich has and demonstrates good safety.
9.Ameliorative Effect of Wendantang Combined with Danshenyin and Dushentang on Ischemic Heart Disease with Phlegm-stasis Syndrome in Mice Based on Circulating Monocytes
Fenghe YANG ; Ziqi TIAN ; Zhiqian SONG ; Shitao PENG ; Wenjie LU ; Tao LIN ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):22-32
ObjectiveTo investigate the ameliorative effect of Wendantang combined with Danshenyin and Dushentang (WDD) on mice with ischemic heart disease (IHD) presenting phlegm-stasis syndrome based on the inflammatory phenotype and differentiation of circulating monocytes. MethodsA model of IHD with phlegm-stasis syndrome was established using left anterior descending coronary artery ligation supplemented with a high-fat diet. Eighty model mice were randomly assigned to the model group, WDD low-dose group (WDD-L), WDD medium-dose group (WDD-M), WDD high-dose group (WDD-H), and atorvastatin calcium tablet group, with 16 mice in each group. An additional 16 C57BL/6J mice were designated as the sham-operation group. The WDD groups received intragastric administration at doses of 8.91, 17.81, 35.62 g·kg-1, and the atorvastatin calcium tablet group received the corresponding drug at 1.3 mg·kg-1, twice daily. The sham-operation and model groups were given the same volume of pure water by gavage each day. After 5 consecutive weeks of administration, the cardiac index was calculated. Cardiac function was assessed by echocardiography. Myocardial histopathology was examined by hematoxylin-eosin (HE) staining. Serum N-terminal pro-B-type natriuretic peptide (pro-BNP) content was measured by enzyme-linked immunosorbent assay (ELISA). Hemorheological parameters were analyzed using an automated hemorheology analyzer. Serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were determined using an automated biochemical analyzer. Changes in circulating monocytes were detected by flow cytometry. Mouse bone marrow mononuclear cells were isolated in vitro and divided into blank group, model serum group, WDD-L drug-containing serum group, WDD-M drug-containing serum group, and WDD-H drug-containing serum group. CD36 expression and macrophage differentiation in each group were assessed by flow cytometry. The mechanism by which WDD mediates circulating monocyte differentiation was further explored using CD36 knockdown/overexpression RAW264.7 cell lines. ResultsCompared with the sham-operation group, the model group showed a significantly increased cardiac index (P<0.01), significantly decreased fractional shortening (FS) (P<0.01), and significantly increased left ventricular end-diastolic internal diameter (LVDD) and left ventricular end-systolic internal diameter (LVDS) (P<0.01). Cardiomyocytes exhibited marked deformation and necrosis with inflammatory cell infiltration. Serum pro-BNP levels were significantly elevated (P<0.01), and whole-blood viscosity (BV) at high, medium, and low shear rates was significantly increased (P<0.01). Compared with the model group, the WDD groups showed significantly reduced cardiac index (P<0.05, P<0.01), significantly increased FS (P<0.05, P<0.01), significantly decreased LVDD and LVDS (P<0.01), markedly improved cardiomyocyte morphology, significantly reduced inflammatory infiltration, significantly decreased serum pro-BNP levels (P<0.01), and significantly decreased BV at high, medium, and low shear rates (P<0.01), with the most pronounced improvement observed in the WDD-M group. Compared with the sham-operation group, TC, TG, and LDL levels were significantly increased in the model group (P<0.05, P<0.01), while HDL levels were significantly decreased (P<0.05). After WDD-H treatment, TC, TG, and LDL levels were significantly reduced and HDL levels were significantly increased in mice (P<0.05, P<0.01). Compared with the sham-operation group, classical monocytes in blood and bone marrow and intermediate monocytes in blood were significantly increased in the model group (P<0.01), whereas intermediate monocytes in bone marrow and non-classical monocytes in blood were significantly decreased (P<0.01). After WDD administration, all circulating monocyte subsets in blood and bone marrow were significantly alleviated (P<0.05, P<0.01), with the WDD-M group showing the optimal effect. In vitro, compared with the blank group, CD36 expression on bone marrow monocytes and the proportion of differentiated macrophages were significantly increased in the model serum group (P<0.01), and CD36 expression was significantly upregulated on RAW264.7 cells (P<0.01). Compared with the model serum group, all drug-containing serum groups exhibited significantly reduced CD36 expression on bone marrow monocytes and significantly reduced macrophage differentiation (P<0.01). WDD downregulated CD36 expression in both CD36 knockdown and overexpression RAW264.7 cell lines (P<0.05, P<0.01), with the strongest regulatory effect observed in the WDD-M drug-containing serum group. ConclusionWDD can significantly improve the manifestations of phlegm-stasis syndrome in IHD mice and reduce the proportion of classical circulating monocytes. Its mechanism may be related to the inhibition of CD36 expression on classical circulating monocytes.
10.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.

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