1.Compatibility Mechanism of Shangerhuang Wan in Enhancing Intestinal Motility in Rat Model of Functional Constipation by Regulating Bile Acid Metabolism
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):1-8
ObjectiveTo explore the mechanism of Shangerhuang Wan (SEHW) and its subdivisions in alleviating loperamide-induced functional constipation (FC) in rats by regulating bile acid metabolism. MethodsSixty rats were randomly assigned into six groups (n=10): normal control, model, positive control (domperidone, 8 mg·kg-1), Cimicifugae Rhizoma-Bupleuri Radix (SC, 1.33 g·kg-1), Scutellariae Radix-Coptidis Rhizoma (QL, 1.20 g·kg-1), and SEHW (3.33 g·kg-1). The remaining groups except the normal control group were subjected to subcutaneous injection with loperamide at 3 mg·kg-1 twice daily (total dose of 6 mg·kg-1·d-1) for 7 days to induce a model of FC. Drug administration was initiated on day 3 of modeling, which continued throughout the modeling period. The small intestinal propulsion rate of each group was measured via the ink propulsion method. LC-MS/MS was employed to measure the fecal bile acid content in each group, and the serum bile acid level was measured by a microplate. The 5-hydroxytryptamine (5-HT) and cyclic adenosine monophosphate (cAMP) levels in the colon tissue of each group were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was employed to determine the protein levels of apical sodium-dependent bile acid transporter (ASBT) in the ileum terminus and Takeda G protein-coupled receptor 5 (TGR5) in the colon. ResultsCompared with the normal control group, the model group exhibited decreases in the small intestinal propulsion rate and total fecal bile acid content (P<0.01), an elevation in the serum total bile acid level, lowered 5-HT and cAMP levels (P<0.01), up-regulation in the protein level of ASBT (P<0.01), and down-regulation in the protein level of TGR5 (P<0.05). Compared with the model group, the positive control, SEHW, and QL groups showed increases in the small intestinal propulsion rate (P<0.05, P<0.01) and total fecal bile acid content (P<0.01) and a decline in the serum bile acid level. Compared with the SEHW group, the SC group had decreased total fecal bile acid content (P<0.01) and an elevated serum bile acid level, while the QL group showed increased total fecal bile acid content (P<0.01) and a lowered serum bile acid level. Compared with the model group, the positive control, SEHW, and QL groups demonstrated down-regulation in the protein level of ASBT (P<0.05, P<0.01) and up-regulation in the protein level of TGR5 (P<0.05, P<0.01). The SC group showed no significant change in the protein level of ASBT and up-regulation in the protein level of TGR5 compared with the model group (P<0.05). Additionally, compared with the model group, the positive control, SEHW, and QL groups showed elevated 5-HT and cAMP levels (P<0.05, P<0.01). The SC group showed no significant difference in the 5-HT level but a rise in the cAMP level (P<0.01). ConclusionSEHW, through the compatibility of QL and SC, demonstrates a dual regulatory effect on bile acid metabolism, embodying the principle of descending turbidity and ascending lucidity, which highlights the compatibility and scientific rationale of this formula. SEHW inhibits ASBT expression in the ileum to reduce ileal bile acid reabsorption, increase colonic bile acid content, and bind to colonic TGR5 to release 5-HT, thereby enhancing intestinal motility and promoting intestinal contraction.
2.Dihuang Yinzi Improves Cognitive Function of Mouse Model of Learning and Memory Impairments by Regulating Synaptic Plasticity via SIRT2
Wenting WANG ; Yangjing HAO ; Wenna SU ; Qinqing LI ; Shifeng CHU ; Junlong ZHANG ; Wenbin HE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):9-17
ObjectiveTo investigate the effects of Dihuang Yinzi on the cognitive function in the mouse model of learning and memory impairments induced by scopolamine (SCOP) and explore the treatment mechanism. MethodsA mouse model of learning and memory impairment was induced by intraperitoneal injection of SCOP. Sixty male C57BL/6J mice were randomized into six groups: control (0.9% NaCl, n=10), model (SCOP 1 mg·kg-1·d-1, n=10), low-, medium-, and high-dose Dihuang Yinzi (SCOP 1 mg·kg-1·d-1 + Dihuang Yinzi 5.5, 11.0, and 22.0 g·kg-1·d-1, n=10), and donepezil (SCOP 1 mg·kg-1·d-1 + donepezil 0.84 mg·kg-1·d-1, n=10). Mice were administrated with corresponding drugs for 6 weeks. Modeling started in the 4th week, and mice in other groups except the control group were injected with SCOP intraperitoneally 40 min after daily gavage. Behavioral testing began in the 5th week, 30 min after modeling each day. The Morris water maze and novel object recognition tests were carried out to evaluate the spatial learning and memory function of mice. Nissl staining was employed to observe the survival of neurons and Nissl bodies in the hippocampal CA1 region. Western blot was employed to determine the protein levels of silent information regulator 2 (SIRT2), α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor 1 (GluA1), protein kinase A (PKA), cAMP response element-binding protein (CREB), phosphorylated-CREB (p-CREB), postsynaptic density protein 95 (PSD95), growth-associated protein-43 (GAP-43), and synaptophysin (SYN) in the hippocampus. Immunofluorescence was used to detect the expression of doublecortin (DCX) in the hippocampal dentate gyrus (DG) region. ResultsCompared with the control group, the model group showed impaired learning and memory (P<0.01), obvious neuronal damage in the hippocampal CA1 region, a reduction in neuron survival (P<0.01), a decrease in DCX expression in the hippocampal DG region (P<0.01), down-regulated proteins levels of GluA1, PKA, p-CREB/CREB, PSD95, SYN, and GAP-43 in the hippocampal tissue (P<0.05, P<0.01), and an up-regulated protein level of SIRT2 (P<0.01). Compared with the model group, the medium- and high-dose Dihuang Yinzi groups and the donepezil group showed improvements in learning and memory (P<0.05, P<0.01), while the low-, medium-, and high-dose Dihuang Yinzi groups and the donepezil group had increased neuron survival (P<0.05, P<0.01). The medium-dose Dihuang Yinzi group and the donepezil group showed increased DCX expression (P<0.05, P<0.01). The medium- and high-dose Dihuang Yinzi groups and the donepezil group showed up-regulation in the protein levels of GluA1, PKA, p-CREB/CREB, PSD95, SYN, and GAP-43 (P<0.05, P<0.01) and down-regulation in the protein level of SIRT2 (P<0.01). ConclusionDihuang Yinzi can improve the cognitive function in the mouse model of learning and memory impairments induced by SCOP by inhibiting the upregulation of SIRT2, activating the PKA/CREB signaling pathway, improving synaptic plasticity, and reducing hippocampal neuronal damage.
3.Xixintang-medicated Serum Regulates Aβ25-35-induced Polarization of BV-2 Microglial Cells
Chaokai YANG ; Yongchang DIWU ; Yangyang WU ; Xia XING ; Dengkun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):18-26
ObjectiveTo investigate the effects of Xixintang (XXT)-medicated serum on the amyloid β-protein (Aβ)25-35-induced polarization of BV-2 microglial cells by a cell experiment and uncover the potential mechanisms of this formula in the prevention and treatment of Alzheimer's disease (AD), thus providing scientific evidence for the clinical application of XXT. MethodsBV-2 microglial cells were subcultured. The optimal concentrations of XXT-medicated serum and Aβ25-35 were determined via the cell-counting kit-8 (CCK-8) assay. The cell experiment was carried out with the following groups: blank control, model (Aβ25-35 at 40 μmol·L-1), XXT-medicated serum (Aβ25-35 at 40 μmol·L-1 + 10% XXT-medicated serum), and blank serum (Aβ25-35 at 40 μmol·L-1 + 10% blank serum). After 24 hours of cell incubation, immunofluorescence was used to detect the expression of ionized calcium-binding adaptor molecule 1 (IBA1), CD16/32, and CD206. Real-time PCR was performed to measure the mRNA levels of CD206, CD163, inducible nitric oxide synthase (iNOS), and arginase 1 (Arg-1). Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the levels of nerve growth factor (NGF), iNOS, and Arg-1. The nitric oxide (NO) concentration was determined via the nitrate reductase method. ResultsCompared with the blank control group, the model group showed increased expression of IBA1 and CD16/32 (P<0.01), decreased expression of CD206 (P<0.05), upregulation in the mRNA level (P<0.01) and content (P<0.05) of iNOS, downregulation in the mRNA levels of CD206, CD163, and Arg-1 (P<0.05, P<0.01), lowered levels of Arg-1 and NGF (P<0.05), and an elevation in the NO level (P<0.05). Compared with the model group, the XXT-medicated serum group exhibited reduced expression of IBA1 and CD16/32 (P<0.05, P<0.01) and increased expression of CD206 (P<0.01). Both the content and mRNA level of iNOS were downregulated (P<0.05, P<0.01), while the mRNA levels of CD206, CD163, and Arg-1 were upregulated (P<0.01) in the XXT-medicated serum group. In addition, the XXT-medicated serum group showed elevated levels of Arg-1 and NGF (P<0.05) and a lowered level of NO (P<0.05). The blank serum group showed no statistically significant differences in the measured parameters compared with the model group. ConclusionThe XXT-medicated serum can inhibit the polarization toward the M1 phenotype and promote the polarization toward the M2 phenotype, exerting anti-inflammatory and neurotrophic effects.
4.Mechanism of Xuefu Zhuyutang in Intervening in Ferroptosis in Rats with Coronary Heart Disease with Blood Stasis Syndrome Based on ACSL4 Signalling Pathway
Yi LIU ; Yang YANG ; Chang SU ; Peng TIAN ; Mingyun WANG ; Ruqian ZHONG ; Xuejiao XIE ; Qing YAN ; Qinghua PENG ; Qiuyan ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):27-38
ObjectiveTo investigate the mechanism of ferroptosis mediated by long-chain acyl-CoA synthetase 4 (ACSL4) signalling pathway in rats with coronary heart disease with blood stasis syndrome and the intervention effect of Xuefu Zhuyutang. MethodsSPF male SD rats were randomly divided into normal group, sham-operation group, model group, trimetazidine group (5.4 mg·kg-1), low-, medium-, and high-dose group (3.51, 7.02,14.04 g·kg-1) of Xuefu Zhuyutang. The coronary artery left anterior descending ligation method was used to prepare a model of coronary heart disease with blood stasis syndrome, and continuous treatment for 7 d was conducted, while the sham-operation group was only threaded and not ligated. The general macroscopic symptoms of the rats were observed, and indicators such as electrocardiogram, echocardiography, and blood rheology were detected. The pathological morphology of myocardial tissue was observed by hematoxylin-eosin (HE) staining, and the changes in mitochondria in myocardial tissue were observed by transmission electron microscopy. The level of iron deposition in myocardial tissue was observed by Prussian blue staining. The levels of 12-hydroxyeicosatetraenoic acid (12-HETE) and 15-HETE were detected in serum by enzyme-linked immunosorbent assay. A biochemical colourimetric assay was used to detect the levels of Fe2+, lipid peroxidation (LPO), glutathione (GSH), and T-GSH/glutathione disulfide (GSSG) in myocardial tissue. DCFH-DA fluorescence quantitative assay was employed to detect the levels of reactive oxygen species (ROS). Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was adopted to detect the protein and mRNA expressions of glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), ACSL4, and ly-sophosphatidylcholine acyltransferase3 (LPCAT3) in myocardial tissue. ResultsCompared with those in the normal group, the rats in the model group were poor in general macroscopic symptoms. The electrocardiogram showed widened QRS wave amplitude and increased voltage, bow-back elevation of the ST segments, elevated T waves, J-point elevation, and accelerated heart rate. Echocardiography showed a significant reduction in left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) (P<0.01). Blood rheology showed that the viscosity of the whole blood (low, medium, and high rate of shear) was significantly increased (P<0.01). HE staining showed an abnormal structure of myocardial tissue. There was a large area of myocardial necrosis and inflammatory cell infiltration and a large number of connective tissue between myocardial fibers. Transmission electron microscopy showed that the mitochondria were severely atrophy or swelling. The cristae were reduced or even broken, and the matrix was flocculent or even vacuolated. Prussian blue staining showed that there were a large number of iron-containing particles, and the iron deposition was obvious. The content of 12-HETE and 15-HETE in the serum was significantly increased (P<0.01). The content of Fe2+, LPO, and ROS in myocardial tissue was significantly increased (P<0.01). The content of GSH was significantly decreased (P<0.01), and T-GSH/GSSG was decreased (P<0.01). The protein and mRNA expressions of GPX4 and FTH1 in myocardial tissue were both significantly decreased (P<0.05, P<0.01), while those of ACSL4 and LPCAT3 increased significantly (P<0.01). Compared with the model group, the general macroscopic symptoms and electrocardiogram results of rats in low-, medium- and high-dose groups of Xuefu Zhuyutang were alleviated, and the differences in LVEF/LVFS ratios were all significantly increased (P<0.05, P<0.01). The differences in whole-blood viscosity (low, medium, and high rate of shear) were all significantly decreased (P<0.01). The results of HE staining and transmission electron microscopy showed that the morphology, structure, and mitochondria of cardiomyocytes were improved. The content of 12-HETE and 15-HETE in serum was reduced to different degrees in low-, medium-, and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01). The content of Fe2+, LPO, and ROS was significantly reduced in the medium- and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01), and the content of GSH and T-GSH/GSSG was significantly increased (P<0.05, P<0.01). The protein and mRNA expressions of GPX4 and FTH1 were significantly increased to varying degrees in the medium- and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01), and ACSL4 and LPCAT3 were decreased to different degrees in the low-, medium-, and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01). ConclusionXuefu Zhuyutang can regulate iron metabolism and anti-lipid oxidation reaction to mediate ferroptosis through the ACSL4 signalling pathway, thus exerting a protective effect on rats with coronary heart disease with blood stasis syndrome.
5.Renshen Yangrongtang Alleviating Myelosuppression by Reducing Neutrophil Extracellular Traps Through Regulating ROS/MPO
Jing ZHANG ; Rongxing LIU ; Jinhao ZENG ; Qing NIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):39-46
ObjectiveTo investigate the potential mechanism of Renshen Yangrongtang in alleviating myelosuppression by regulating the expression of reactive oxygen species (ROS), myeloperoxidase (MPO), and neutrophil extracellular traps (NETs). MethodsK562 cells were divided into blank group, etoposide group (40 μmol·L-1), and etoposide+Renshen Yangrongtang freeze-dried powder groups with low-, medium-, and high-dose (2, 4, 8 g·L-1). Liquid chromatography-mass spectrometry (LC-MS) was used to determine the freeze-dried powder of Renshen Yangrongtang. Enzyme-Linked Immunosorbent Assay (ELISA) was used to detect ROS, MPO, and NETs expression in each group. Western blot analysis was performed to assess intracellular MPO and NE expressions. Twenty 8-week-old male mice were randomly divided into blank group, etoposide group (100 mg·kg-1), and etoposide + Renshen Yangrongtang groups with low-, medium-, and high-dose (0.1, 0.5, 2.0 g·kg-1). Except for the blank group that received PBS via gavage at room temperature, and the etoposide group that received an intraperitoneal injection for 3 days, the remaining groups received gavage of Renshen Yangrongtang for 14 consecutive days after 3 days of etoposide administration. The peripheral blood related indicators were detected through an automated hematology analyzer; Western blot analysis was performed to assess MPO and neutrophil elastase (NE) expression changes in the marrow cells of mice. Enzyme-linked immunosorbent assay (ELISA) was used to detect ROS, MPO, and NETs changes in the marrow cells of mice. MPO and NE on femur bones were stained through immunohistochemistry. Scanning electron microscopy was used to analyze the structural changes of NETs in the marrow cells of mice after drug administration. ResultsLC-MS results showed that the freeze-dried powder of Renshen Yangrongtang contained complete technical materials such as Chinese angelica, Astragalus mongholicus, and ginseng. In K562 cells, compared with the etoposide group, ELISA results indicated that the concentrations of MPO, ROS, and NETs in the etoposide + Renshen Yangrongtang medium and high-dose groups were decreased (P<0.05, P<0.01), and Western blot data showed that the etoposide high-dose group significantly reduced the expression of MPO and NE protein in K562 cells (P<0.05, P<0.01). In vivo, compared with the etoposide group, the number of RBC, WBC, and PLT in the etoposide+Renshen Yangrongtang high-dose group increased significantly (P<0.05). ELISA results suggested that in the etoposide+Renshen Yangrongtang low-, medium-, and high-dose groups, the concentration of mice ROS, MPO, and NETs significantly decreased (P<0.05, P<0.01). Western blot results revealed that compared with the etoposide group, the expressions of MPO and NE in the marrow cells of mice in the etoposide + Renshen Yangrongtang low-, medium- and high-dose groups were significantly decreased (P<0.05, P<0.01). Scanning electron microscopy observations revealed that Renshen Yangrongtang reduced the NETs structure generation in the marrow cells of mice after the influence of etoposide. ConclusionRenshen Yangrongtang can alleviate etoposide-induced myelosuppression by inhibiting ROS/MPO and reducing the formation of intracellular NETs.
6.Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-dampness Obstruction Syndrome and Dampness-heat Obstruction Syndrome
Na LIN ; Yanqiong ZHANG ; Changhong XIAO ; Shenghao TU ; Jianning SUN ; Shijun XU ; Representation Preparation GROUP
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):47-54
Rheumatoid arthritis belongs to arthralgia syndrome in the theory of traditional Chinese medicine, and cold-dampness obstruction syndrome and dampness-heat obstruction syndrome are core syndromes and main syndrome differentiation types of this disease. Fine therapeutic effects have been obtained in the long-term clinical practice of many famous traditional Chinese medicine practitioners following the syndrome differentiation and treatment based on the guiding principles of cold and heat. To adapt to the clinical diagnosis practice of combining disease differentiation and syndrome differentiation, and to better carry out basic research on integrated Chinese and Western medicine and preclinical study on new traditional Chinese medicines, Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-Dampness Obstruction Syndrome and Dampness-Heat Obstruction Syndrome (hereinafter referred to as the Guidelines) were compiled by our research group, in cooperation with the renowned experts in research fields including traditional Chinese medicine, clinical medicine, zoology and evidence-based medicine, which provide a meaningful reference for scientific research, teaching and clinical applications. The compilation process of the Guidelines was guided by the theory of disease and syndrome integration and the principles of "evidence takes the main place, consensus plays an auxiliary role, and experience serves as the reference". Based on the comprehensive evaluation of pathogenesis homology, behavioral phenotypic consistency, and drug treatment predictability compared between animal models and human diseases, by the nominal group method, "recommendations" were formed for recommendations supported by evidence, and "consensus recommendations" were formed for recommendations not supported by evidence. Guidelines were formed involving content such as animal types, arthritis modeling methods, external stimulation conditions, and modeling assessment indicators during the establishment of the animal models of rheumatoid arthritis with cold-dampness obstruction syndrome and dampness-heat obstruction syndrome. The Guidelines are applicable for the disease and syndrome research on rheumatoid arthritis, investigation of therapeutic mechanisms, and development of new traditional Chinese medicine. The Guidelines also provide a reference for the establishment of guidelines on other types of diseases and syndromes combined with animal models to further promote the modernization of traditional Chinese medicine research and its integration with international academic development.
7.Editorial Explanation of Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-dampness Obstruction Syndrome and Dampness-heat Obstruction Syndrome
Na LIN ; Yanqiong ZHANG ; Changhong XIAO ; Shenghao TU ; Jianning SUN ; Shijun XU ; Xia MAO ; Representation Preparation GROUP
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):55-59
The Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-dampness Obstruction Syndrome and Dampness-heat Obstruction Syndrome (hereinafter referred to as the Guidelines) (No. T/CACM1567-2024) was published by Chinese Association of Chinese Medicine on January 11, 2024. To assist researchers and medical workers in understanding and applying the Guidelines more accurately, and also to provide reference and assistance for the establishment of guidelines on other types of diseases and syndromes combined with animal models, this paper made a declaration of the workflow, technological links, development references, promotion of its application and after-effect evaluation of the Guidelines that has been made according to the requirements of "Draft Group Standard of the Standardization Office of the Chinese Association of Traditional Chinese Medicine".
8.Mechanism of Intervening with Diarrhea-predominant Irritable Bowel Syndrome in Rats with Spleen Deficiency by Xingpi Capsules Through Regulating 5-HT-RhoA/ROCK2 Pathway
Gang WANG ; Lingwen CUI ; Xiangning LIU ; Rongxin ZHU ; Mingyue HUANG ; Ying SUN ; Boyang JIAO ; Ran WANG ; Chun LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):60-69
ObjectiveTo investigate the efficacy of Xingpi capsules (XPC) in treating diarrhea-predominant irritable bowel syndrome (IBS-D) with spleen deficiency and elucidate its potential molecular mechanisms. MethodsA rat model of IBS-D with spleen deficiency was established by administering senna leaf in combination with restrained stress and swimming fatigue for 14 d. Ten specific pathogen free (SPF)-grade healthy rats were used as the normal control group. After successful modeling, SPF-grade rats were randomly divided into a model group, a pinaverium bromide group (1.5 mg·kg-1), and low- and high-dose XPC groups (0.135 and 0.54 g·kg-1), with 10 rats in each group. Rats in the normal control group and the model group were given distilled water by gavage, while the remaining groups were administered corresponding drug solutions by gavage once a day for 14 consecutive days. The rat body weights and fecal condition were observed every day, and the Bristol score was recorded. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of 5-hydroxytryptamine (5-HT) in serum and colon tissue. Transmission electron microscopy was used to observe the microvilli and tight junctions in the colon. The integrity of the colonic barrier, intestinal motility, and expression of related pathway proteins were evaluated by hematoxylin-eosin (HE) staining, immunohistochemistry, and Western blot. ResultsCompared with those in the normal control group, rats in the model group showed a significantly decreased body weight and increased diarrhea rate, diarrhea grade, and Bristol score (P<0.01). HE staining revealed incomplete colonic mucosa in the model group, with evident congestion and edema observed. Electron microscopy results indicated decreased density and integrity of the colonic barrier, shedding and disappearance of microvilli, and significant widening of tight junctions. The expression levels of colonic tight junction proteins Occludin and Claudin-5 were downregulated (P<0.01), and the levels of 5-HT in serum and colon tissue were elevated (P<0.01). The small intestine propulsion rate significantly increased (P<0.01), and the expression of contractile proteins Ras homolog family member A (RhoA) and Rho-associated coiled-coil containing protein kinase 2 (ROCK2) in colon and phosphorylation of myosin light chain (MLC20) were upregulated (P<0.01). Compared with the model group, the treatment groups showed alleviated diarrhea, diarrhea-associated symptoms, and pathological manifestations of colon tissue to varying degrees. Specifically, high-dose XPC exhibited effectively relieved diarrhea, promoted recovery of colonic mucosal structure, significantly reduced congestion and edema, upregulated expression of Occludin and Claudin-5 (P<0.01), decreased levels of 5-HT in serum and colon tissue (P<0.05,P<0.01), significantly slowed small intestine propulsion rate (P<0.01), and significantly downregulated expression of contractile proteins RhoA and ROCK2 in colon and phosphorylation of MLC20 (P<0.05,P<0.01). ConclusionXPC effectively alleviates symptoms of spleen deficiency and diarrhea and regulates the secretion of brain-gut peptide. The characteristics of XPC are mainly manifested in alleviating IBS-D with spleen deficiency from the aspects of protecting intestinal mucosa and inhibiting smooth muscle contraction, and the mechanism is closely related to the regulation of the 5-HT-RhoA/ROCK2 pathway expression.
9.Effect of Qingxin Jieyu Granules Regulating Mitophagy on Ventricular Remodeling After Myocardial Infarction of C57B/L6 Mice
Yifan CHEN ; Jianfeng CHU ; Zhonghui JIANG ; Zhuye GAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):70-78
ObjectiveTo explore the effects of Qingxin Jieyu granules on ventricular remodeling of mice after myocardial infarction, and their regulatory role in mitophagy. MethodsSixty male C57BL/6 mice were randomly assigned to six groups: sham-operated group, model group, Qingxin Jieyu granules low-, medium-, and high-dose groups (1.3, 2.6, 5.2 g·kg-1), and sacubitril valsartan sodium group (0.03 g·kg-1), with 10 mice per group. Except for the sham-operated group, all other groups utilized left anterior descending coronary artery ligation to build a myocardial infarction model. Ultrasound was used to measure left ventricular parameters, including end-diastolic and end-systolic diameters (LVIDd, LVIDs), diastolic and systolic posterior wall thickness (LVPWd, LVPWs), end-diastolic and end-systolic volumes (LV Vold, LV Vols), left ventricular ejection fraction (LVEF), and fractional shortening (LVFS). Additionally, the heart mass index and heart weight/tibia length ratio of mice were calculated. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify brain natriuretic peptide (BNP), creatine kinase isoenzyme (CK-MB), angiotensinⅡ (AngⅡ), and lactate dehydrogenase (LDH) levels in the serum of mice. Histological analysis using hematoxylin-eosin (HE) and Masson staining was conducted to examine morphological changes in myocardial tissue. Immunohistochemistry assessed the expression of vascular growth factors, including basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF). Transmission electron microscopy was used to scrutinize mitochondrial morphology in the myocardial tissue of mice. Western blot was performed to analyze the expression of phosphorylated adenosine monophosphate activated protein kinase (p-AMPK) and phosphorylated mammalian target of rapamycin (p-mTOR) proteins in myocardial tissue from each experimental group. ResultsCompared to the sham-operated group, the model group mice exhibited significantly elevated levels of LV Vold, LV Vols, LVIDd, LVIDs, cardiac mass index, heart weight/tibia length ratio, BNP, LDH, and p-mTOR protein expression (P<0.05), along with decreased levels of LVPWd, LVPWs, LVEF, LVFS, and p-AMPK protein expression (P<0.05). The model group also displayed substantial inflammatory cell infiltration, collagen deposition in myocardial cells, reduced expression of bFGF and VEGF, mitochondrial swelling, and cristae fragmentation. Compared to the model group, the sacubitril/valsartan group and mid-dose Qingxin Jieyu granules group showed significant reductions in LVIDs, LV Vold, LV Vols, BNP, CK-MB, LDH, and p-mTOR protein expression (P<0.05), coupled with increases in LVEF, LVFS, and p-AMPK expression (P<0.05). Improvements were observed across all treatment groups, including reduced inflammatory cell infiltration and collagen deposition, increased bFGF and VEGF expression, alleviated mitochondrial swelling, and the presence of autophagosomes and lysosomes
10.Efficacy Mechanism of Xianlian Jiedu Prescription Against Colorectal Cancer Recurrence vias Regulating Angiogenesis
Yanru XU ; Lihuiping TAO ; Jingyang QIAN ; Weixing SHEN ; Jiani TAN ; Chengtao YU ; Minmin FAN ; Changliang XU ; Yueyang LAI ; Liu LI ; Dongdong SUN ; Haibo CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):79-87
ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer (CRC) and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit, followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group, low-dose Xianlian Jiedu prescription (XLJDP-L) group (6.45 g·kg-1·d-1), medium-dose Xianlian Jiedu prescription (XLJDP-M) group (12.9 g·kg-1·d-1), high-dose Xianlian Jiedu prescription (XLJDP-H) group (25.8 g·kg-1·d-1), and 5-fluorouracil (5-FU) group (1×10-3 g·kg-1·d-1). The mice were euthanized after 14 days of continuous intervention, and recurrent tumor tissue was harvested. Hematoxylin and eosin (HE) staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry (IHC) was employed to assess the expression of proliferating cell nuclear antigen (Ki67), vascular endothelial growth factor (VEGF), and platelet-endothelial cell adhesion molecule (CD31) in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 (ANG-2), VEGF, phosphorylated-protein kinase B (p-Akt), protein kinase B (Akt), phosphorylated-phosphatidylinositol 3-kinase (p-PI3K), and phosphatidylinositol 3-kinase (PI3K) in recurrent tumor tissue. ResultsBefore treatment, there were no statistical differences in tumor volume, tumor weight, and body mass among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group, indicating model stability. After treatment, compared with those in the model group, the tumor volume and tumor weight in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01), showing dose dependency. Meanwhile, there were no significant differences in body weight among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group, tumor tissue in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group had loosely arranged cells, increased intercellular spaces, small and shriveled nuclei, light staining, fewer mitotic figures and atypical nuclei, and increased necrotic areas. IHC showed that compared with those of the model group, the positive rates of Ki67, VEGF, and CD31 in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01) in a dose-dependent manner. Western blot results showed that compared with those of the model group, the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly downregulated (P<0.05, P<0.01), and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner (P<0.05, P<0.01). ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2, VEGF, and CD31.

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