1.Based on 16S rDNA Technology and TLRs/MyD88/NF-κB Signaling Pathway, Molecular Mechanism of Shenling Baizhusan Resistance to Diarrhea Irritable Bowel Syndrome Rats Was Investigated
Tengfei LYU ; Jingyu WANG ; Mingyue XIE ; Bin XI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):13-22
ObjectiveBased on 16S rDNA technology and molecular biology methods, the molecular mechanism of Shenling Baizhusan in the treatment of diarrhea-predominant irritable bowel syndrome (IBS-D) was investigated. MethodsThe 42 SD rats with SPF were randomly divided into no control group, SLBZS-H, medium (SLBZS-M), low (SLBZS-L) dose group, positive control group and model group, with 7 rats in each group. The rat model of IBS-D was prepared by ice-cold senna (0.45 g∙mL-1) gavage (10 mL∙kg-1) combined with restraint stress for 14 consecutive days. After successful modeling, the corresponding drugs were given to each group with a gavage volume of 10 mL∙kg-1: The positive group was administered with 2.36 , 1.18, 0.59 g∙mL-1 of Shenling Baizhusan in the Positive group and the Model group with the same volume of normal saline for 14 d. The general condition of the rats: Weight, feces, mental state and death were observed and recorded. The body weight, abdominal wall retraction reflex score (AWR) and loose stool rate of rats in each group were measured before (the first day), after the model (day 14) and after treatment (day 28). Hematoxylin-eosin staining was used to observe the morphological characteristics of colon tissues of experimental animals. Enzyme-linked immunosorbent assay was used to quantitatively analyze the concentration of inflammatory mediators in the peripheral blood of experimental animals. Western blotting was used to detect the expression levels of key proteins of Toll-like receptor 4 (TLR4), Toll-like receptor 2 (TLR2), myeloid differentiation factor 88 (MyD88) and nuclear factor-κB (NF-κB) signaling pathway in rat colon tissue. 16S rDNA technology was used to detect the structural changes of intestinal microbiota in rats. ResultsCompared with Control, the colon of the Model group showed partial mucosal epithelial shedding and inflammatory cell infiltration. The contents of TNF-α, IL-1β, IL-6 and 5-HT in serum increased (P<0.05), the protein expressions of TLR2, TLR4, MyD88 and NF-κB in colon tissue increased (P<0.05), the diversity indices of Richness, Chao1, abundance-based coverage estimator(ACE) and Shannon decreased (P<0.05), and the phylum Firmicutes, Actinobacteria, The relative richness of Bacteroides-H, Lactobacillus and Ligilactobacillus decreased (P<0.05), while the relative richness of Bacteroidetes, Proteobacteria and Prevotella increased (P<0.05). Compared with the model group, the colonic structure and organization of the SLBZS-H group, SLBZS-M group, SLBZS-L group and Positive group were clearer, and only a small number of inflammatory cells were present in some areas, and the serum contents of TNF-α, IL-1β, IL-6 and 5-HT were decreased (P<0.05), TLR2, TLR4, The protein expressions of MyD88 and NF-κB decreased (P<0.05), and compared with the model group, the diversity indices of Richness, Chao1, ACE and Shannon in the SLBZS-H, SLBZS-M and SLBZS-L groups increased (P<0.05), and the richness of Firmicutes and Actinobacteria increased (P<0.05). The richness of Proteobacteria and Prevotella decreased (P<0.05), and the abundance of Prevotella decreased (P<0.05), Bacteroides-H, Muribaculum, Lactobacillus and salivarius in the Positive group salivarius (P<0.05). ConclusionShenling Baizhusan can effectively treat IBS-D, and its molecular mechanism may be to play a therapeutic role by improving intestinal flora and inhibiting the TLRS/MyD88/NF-κB signaling pathway to reduce inflammatory response.
2.Exploring Mechanism of Modified Danggui Yinzi in Regulating "Itch-anxiety" Cycle of Chronic Urticaria Based on STEP/NR2B Signaling Pathway
Mingyue LI ; Xinyu XIAO ; Anjing CHEN ; E LIU ; Xurui WANG ; Qin ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):123-133
ObjectiveTo explore the effects and mechanism of the modified Danggui Yinzi on "itch-anxiety" model rats of chronic urticaria (CU). MethodsThe 36 SPF-grade 6-8-week-old female SD rats were randomly divided into a blank control group,a model group,a positive control group,a low-dose modified Danggui Yinzi group,a medium-dose modified Danggui Yinzi group,and a high-dose modified Danggui Yinzi group. A "itch-anxiety" model was established by intraperitoneal injection of a suspension of sodium chloride and aluminum hydroxide and ovalbumin,combined with chronic unpredictable emotional stress stimulation. After successful modeling,rats in each group were administered drugs by gavage. The positive control group was given intragastric administration of the drug solutions of cetirizine and fluoxetine (2.08 mg·kg-1·d-1 fluoxetine, 2 mg·kg-1·d-1 cetirizine), the low-,medium-,and high-dose modified Danggui Yinzi groups were administered traditional Chinese medicine at 1.44,2.88, 5.76 g·kg-1, respectively,while the blank control group and model group were given an equal volume of normal saline. All interventions lasted for 15 days. Behavioral changes were evaluated by the elevated plus-maze test (detecting the percentage of entries into the open arms (OE%),the percentage of time spent in the open arms (OT%),and the total number of entries into the open and closed arms (TNE)),the open-field test (detecting total activity,average movement speed,and latency to enter the central area),and scratching behavior observation. Pathological changes of skin tissues were observed by hematoxylin-eosin (HE) staining and toluidine blue staining,while those of amygdala tissues were observed by HE staining,Nissl staining,and immunofluorescence detection of ionized calcium-binding adapter molecule-1 (Iba-1). The content of immunoglobulin E (IgE),interleukin-33 (IL-33),histamine in serum and glutamate in the amygdala was detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the protein expression of striatal-enriched protein tyrosine phosphatase (STEP),N-methyl-D-aspartate receptor subunit 2B (NR2B), calmodulin-dependent protein kinase Ⅱ (CaMKⅡ),phosphorylated CaMKⅡ (p-CaMKⅡ),mitogen-activated protein kinase (MAPK),phosphorylated MAPK (p-MAPK),nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB),phosphorylated NF-κB (p-NF-κB),and postsynaptic density protein-95 (PSD-95) in the amygdala. ResultsCompared with the blank control group,the model group rats showed obvious anxiety-like behaviors (decreased OE%,OT%,and TNE,reduced total activity,slower average movement speed,and prolonged latency to enter the central area),increased scratching times,obvious skin inflammation and mast cell degranulation,severe amygdala tissue damage,increased glutamate content in the amygdala,and elevated levels of IgE and IL-33 in serum. The expression of STEP,NF-κB,p-NF-κB,NR2B,MAPK,p-MAPK,CaMKⅡ,and p-CaMKⅡ proteins in the amygdala increased,while the expression of PSD-95 protein decreased (P<0.05). Compared with the model group,the modified Danggui Yinzi group of each dose had increased OE%,OT%,TNE,total activity,and average movement speed,shortened latency to enter the central area, reduced scratching times,alleviated skin inflammation and mast cell degranulation,relieved amygdala tissue damage,decreased glutamate content in the amygdala,and reduced levels of IgE and IL-33 in serum. Moreover,compared with the model group,the low -,medium-,and high-dose modified Danggui Yinzi groups showed decreased expression levels of STEP,NF-κB,p-NF-κB,NR2B,MAPK,p-MAPK,CaMKⅡ,and p-CaMKⅡ proteins in the amygdala,and increased expression of PSD-95 protein. There was a significant dose-effect relationship,with the high-dose group showing the most significant regulatory effect (P<0.05). ConclusionThe modified Danggui Yinzi has a therapeutic effect on "itch-anxiety" model rats of CU. Its mechanism may be related to regulating glutamate metabolism in the amygdala,modulating the STEP/NR2B/CaMKⅡ/MAPK/NF-κB pathway,and regulating the expression of PSD-95.
3.Evaluate the effect of morphology driven preparation technique in indirect restoration of posterior teeth
Qikun GAO ; Shiming LIU ; Mingyue WU ; Xiaoting WU ; Zehua WANG ; Mingliang DU ; Huimin CHEN
Acta Universitatis Medicinalis Anhui 2026;61(5):943-947
ObjectiveTo evaluate the application effect of morphology driven preparation technique in indirect restoration of posterior teeth. Methods84 patients with dental defects were selected and divided into the control group and the experimental group randomly, with 42 patients in each group (a total of 84 teeth). Traditional preparation and morphology driven preparation techniques were used to complete dental preparation and adhesive glass ceramic restorations, respectively. The satisfaction, masticatory function, World Dental Federation (FDI) Scores for edge fracture and fixation,edge adaptability index,periodontal bleeding index (BI) and plaque index (PLI) of the two groups of patients were evaluated and various indicator data were recorded and statistically analyzed at 3 and 6 months after restoration completion. ResultsThere was no statistically significant difference in satisfaction between the two groups after 3 and 6 months of repair; Both groups of data showed a significant improvement in masticatory function. There was no statistical difference after 3 and 6 months of repair. However, a statistically significant difference in masticatory function was noted after 6 months of repair compared with before and after 3 months of repair. As the repair time increased, the masticatory effect improved significantly. There was no statistically significant difference in the edge fracture and fixation index between the two groups at 3 months after repair, but there was a statistically significant difference between the two groups at 6 months with the experimental group outperforming the control group(P<0.05). The edge adaptability, BI, and PLI index were statistically significant between the two groups after 3 and 6 months of repair and the experimental group had a significant advantage (P<0.05). ConclusionBoth types of tooth preparation techniques can improve patient masticatory function and reach high satisfaction after repair. However, morphology driven preparation technique has significant advantages in edge adaptability, edge fracture and fixation index scores and improve periodontal conditions. This technique is an effective method for improving dental preparation in adhesive indirect restoration of posterior teeth.
4.Effects and mechanisms of modified Xiaoqinglong decoction combined with BMSCs on nasal-pulmonary-intestinal mucosal immunity in cold asthma rats
Mingyue REN ; Guihua SONG ; Tingting WANG ; Mengmeng SUN ; Bingxue ZHANG ; Minghao PENG
China Pharmacy 2026;37(13):1691-1696
OBJECTIVE To investigate the effects and mechanism of modified Xiaoqinglong decoction combined with bone marrow mesenchymal stem cells (BMSCs) on nasal-pulmonary-intestinal mucosal immunity in rats with cold asthma. METHODS SD rats were randomly divided into blank group, model group, BMSCs group, BMSCs+modified Xiaoqinglong decoction (6.39 g/kg) group, and BMSCs+dexamethasone (0.125 mg/kg) group, with 10 rats in each group. The cold asthma rat model was established by sensitization and atomization challenge with ovalbumin combined with aluminum hydroxide, together with cold air stimulation from air conditioners. One hour before the initial atomization challenge, rats in all groups except the blank group and model group were injected with BMSCs suspension via the tail vein. Starting from the 15th day of the experiment, rats in each group were intragastrically administered corresponding liquid medicine or normal saline once a day for 7 consecutive days. After the last administration, the asthmatic behavioral changes and pathological morphological changes of lung tissues in rats were evaluated. The levels of interleukin-6 (IL-6) and IL-10 in nasopharyngeal lavage fluid, bronchoalveolar lavage fluid (BALF) and intestinal mucus were detected. The expression levels of proteins related to the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in nasal, lung and intestinal tissues were determined. RESULTS Compared with the blank group, the asthmatic behavioral score, lung tissue injury score, IL-6 levels in nasopharyngeal lavage fluid, BALF and intestinal mucus, PI3K protein expression and Akt phosphorylation levels in nasal, lung and intestinal tissues of rats in the model group were significantly increased ( P <0.05); the IL-10 levels in nasopharyngeal lavage fluid, BALF and intestinal mucus were significantly decreased ( P <0.05). Alveolar atrophy, collapse and even fusion, abnormal hyperplasia and hypertrophy of bronchial smooth muscle, and massive inflammatory cell infiltration were observed. Compared with the model group, most of the above quantitative indicators in the BMSCs group, BMSCs+modified Xiaoqinglong decoction group and BMSCs+dexamethasone group were significantly reversed ( P <0.05), and pathological injuries of lung tissues were markedly alleviated. Compared with the BMSCs group, most of the above quantitative indicators were further improved in the BMSCs+modified Xiaoqinglong decoction group ( P <0.05). CONCLUSIONS Modified Xiaoqinglong decoction combined with BMSCs can effectively improve nasal-pulmonary-intestinal mucosal immunity and relieve asthmatic symptoms in cold asthma rats, and its mechanism may be related to inhibiting the activation of the PI3K/Akt signaling pathway.
5.Effects and mechanisms of modified Xiaoqinglong decoction combined with BMSCs on nasal-pulmonary-intestinal mucosal immunity in cold asthma rats
Mingyue REN ; Guihua SONG ; Tingting WANG ; Mengmeng SUN ; Bingxue ZHANG ; Minghao PENG
China Pharmacy 2026;37(13):1691-1696
OBJECTIVE To investigate the effects and mechanism of modified Xiaoqinglong decoction combined with bone marrow mesenchymal stem cells (BMSCs) on nasal-pulmonary-intestinal mucosal immunity in rats with cold asthma. METHODS SD rats were randomly divided into blank group, model group, BMSCs group, BMSCs+modified Xiaoqinglong decoction (6.39 g/kg) group, and BMSCs+dexamethasone (0.125 mg/kg) group, with 10 rats in each group. The cold asthma rat model was established by sensitization and atomization challenge with ovalbumin combined with aluminum hydroxide, together with cold air stimulation from air conditioners. One hour before the initial atomization challenge, rats in all groups except the blank group and model group were injected with BMSCs suspension via the tail vein. Starting from the 15th day of the experiment, rats in each group were intragastrically administered corresponding liquid medicine or normal saline once a day for 7 consecutive days. After the last administration, the asthmatic behavioral changes and pathological morphological changes of lung tissues in rats were evaluated. The levels of interleukin-6 (IL-6) and IL-10 in nasopharyngeal lavage fluid, bronchoalveolar lavage fluid (BALF) and intestinal mucus were detected. The expression levels of proteins related to the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in nasal, lung and intestinal tissues were determined. RESULTS Compared with the blank group, the asthmatic behavioral score, lung tissue injury score, IL-6 levels in nasopharyngeal lavage fluid, BALF and intestinal mucus, PI3K protein expression and Akt phosphorylation levels in nasal, lung and intestinal tissues of rats in the model group were significantly increased ( P <0.05); the IL-10 levels in nasopharyngeal lavage fluid, BALF and intestinal mucus were significantly decreased ( P <0.05). Alveolar atrophy, collapse and even fusion, abnormal hyperplasia and hypertrophy of bronchial smooth muscle, and massive inflammatory cell infiltration were observed. Compared with the model group, most of the above quantitative indicators in the BMSCs group, BMSCs+modified Xiaoqinglong decoction group and BMSCs+dexamethasone group were significantly reversed ( P <0.05), and pathological injuries of lung tissues were markedly alleviated. Compared with the BMSCs group, most of the above quantitative indicators were further improved in the BMSCs+modified Xiaoqinglong decoction group ( P <0.05). CONCLUSIONS Modified Xiaoqinglong decoction combined with BMSCs can effectively improve nasal-pulmonary-intestinal mucosal immunity and relieve asthmatic symptoms in cold asthma rats, and its mechanism may be related to inhibiting the activation of the PI3K/Akt signaling pathway.
6.Mechanism of Shexiang Tongxin Dripping Pills in Ameliorating AngⅡ-induced Vascular Smooth Muscle Cell Dysfunction
Yueyao HU ; Wei WANG ; Mingyue HUANG ; Shujuan GUO ; Junkai YAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):97-106
ObjectiveTo study the mechanism of Shexiang Tongxin Dripping pills-containing serum (STDP) in ameliorating angiotensinⅡ (AngⅡ)-induced cell phenotype transformation, proliferation, migration, and dysfunction of vascular smooth muscle cells. MethodsAn AngⅡ-induced proliferation and migration model of vascular smooth muscle cells was established. The cells were treated with STDP at 5%, 10%, and 20% for 24 h. The immunofluorescence assay was employed to detect the expression of α smooth muscle actin (α-SMA) and matrix metalloproteinase-2 (MMP-2). The cell-counting kit-8 (CCK-8) assay and 5-ethynyl-2'-deoxyuridine (EdU) staining were employed to detect the proliferation of vascular smooth muscle cells, and the scratch assay was employed to detect the migration of the cells. Western blot was employed to determine the expression levels of pathway proteins such as angiotensin-converting enzyme 2 (ACE2), angiotensin Ⅱ type 2 (AT2), angiotensin Ⅱ type 1 (AT1), as well as proliferation and migration proteins such as typeⅠ collagen (ColⅠ) and osteopontin (OPN). ResultsCompared with the model group, STDP increased the expression of α-SMA, reduced the expression of MMP-2, and inhibited the proliferation and migration of vascular smooth muscle cells (P<0.05). Furthermore, STDP up-regulated the expression levels of ACE2, AT2, and MAS1, while down-regulating the expression level of AT1, PCNA, ColⅠ, MMP-9, Rock1, Rock2, and SRF (P<0.05). Compared with the STDP group, the ACE2 inhibitor reversed the regulatory effects of STDP. ConclusionSTDP inhibits the phenotype transformation, proliferation, and migration of vascular smooth muscle cells and regulates the expression of cell proliferation and migration-related proteins to ameliorate the dysfunction of vascular smooth muscle cells. It exerts the effects by up-regulating the expression of proteins in the ACE2-AT2/MAS pathway and down-regulating the expression of proteins in the AT1-Rock signaling pathway.
7.Protective effects of exosomes derived from MSCs in radiation-induced lung injury
Lili WANG ; Zien YANG ; Mingyue OUYANG ; Sining XING ; Song ZHAO ; Huiying YU
Chinese Journal of Radiological Health 2025;34(1):13-20
Objective To investigate the role and related mechanisms of exosomes derived from mesenchymal stem cells (MSCs) in radiation-induced lung injury (RILI). Methods Human umbilical cord-derived MSCs were isolated and cultured for the extraction and identification of exosomes. Eighteen male SD rats were randomly divided into Control group, RILI group and RILI + exosomes group (EXO group), with 6 rats in each group. Except for Control group, the other groups received a single X-ray dose of 30 Gy to the right lung. Immediately after irradiation, the EXO group was administered 2 × 109 exosomes/kg via tail vein injection. Control group and RILI group were given the same volume of normal saline. Eight weeks post-irradiation, the rats were sacrificed, lung tissue and peripheral venous blood were collected. HE and Masson staining were employed to observe the pathological and fibrotic changes of lung tissue. The levels of serum inflammatory factors IL-6, IFN-γ, TNF-α, and IL-10 were detected by ELISA. RT-qPCR was used to assess the mRNA levels of IL-1β, IL-6, Cdh1, and Col1a1 in lung tissue. The expression levels of Vimentin and TGF-β1 in lung tissue were measured by immunohistochemical staining. The expression levels of AMPK, p-AMPK, and TGF-β1 in lung tissue were detected by Western blot. Results MSC-derived exosomes were successfully extracted and identified. Compared with RILI group, EXO group showed significantly reduced pathological changes of lung inflammation and collagen deposition. The levels of serum inflammatory factors IL-6, INF-γ, and TNF-α were significantly decreased (P < 0.05), and the level of anti-inflammatory factor IL-10 was significantly increased (P < 0.05). The mRNA levels of IL-1β, IL-6, and Col1a1 in lung tissue were significantly decreased (P < 0.05 or P < 0.01), and the mRNA level of Cdh1 was significantly increased (P < 0.05 or P < 0.01). The levels of Vimentin and TGF-β1 in lung tissue were significantly reduced, while p-AMPK level was significantly up-regulated (P < 0.05). Conclusion Exosomes derived from MSCs may alleviate RILI by inhibiting inflammatory responses and regulating epithelial-mesenchymal transition mediated by AMPK/TGF-β1 signaling pathway.
8.Effects of parthenolide on systemic inflammation and intestinal injury in rats with acute pancreatitis
Yanan ZHAO ; Rui ZHANG ; Shuling WANG ; Chunchun YANG ; Yang WANG ; Mingyue YANG
China Pharmacy 2025;36(6):704-709
OBJECTIVE To investigate the effects of parthenolide (PLT) on systemic inflammation and intestinal injury in rats with acute pancreatitis (AP) by regulating the Kelch-like epichlorohydrin-associated protein 1 (Keap1)/nuclear factor-erythroid-2 related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway. METHODS AP rat model was established by injecting 3.5% sodium taurine cholate solution (1 mL/kg) into the biliary pancreatic duct, and modeled rats were divided into AP group, PLT (300 µg/kg) group, dexamethasone (2 mg/kg) group, inhibitor (11 mg/kg Nrf2 inhibitor ML385) group, and PLT+inhibitor group (300 µg/kg PLT+11 mg/kg ML385), with 10 rats in each group. Another 10 rats were taken as a sham operation group. Each group was given relevant medicine or normal saline via tail vein/intraperitoneal injection once. After 24 h, serum lipase and amylase levels, the levels of oxidative stress index [superoxide dismutase (SOD) and malondialdehyde (MDA)] and inflammatory factors [interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α)] were detected. The histopathological changes in colon mucosa and pancreas were observed, and Chiu and Schmidt scores were performed. The cell apoptosis in colon mucosa and the protein expressions of Keap1, Nrf2 and HO-1 were detected. RESULTS Compared with the sham operation group, there was obvious inflammatory cell infiltration in colon mucosa and pancreatic tissue, cell shedding or tissue necrosis and severe bleeding; serum levels of lipase, amylase, MDA, IL-1β, IL-6 and TNF-α, Chiu and Schmidt scores, apoptotic rate and protein expression of Keap1 in colonic mucosa were significantly increased or up-regulated, while SOD level and protein expressions of Nrf2 20230993) and HO-1 were decreased or down-regulated significantly (P<0.05). Compared with the AP group, the above indexes in the PLT group and dexamethasone group were significantly improved, while those in the inhibitor group further deteriorated (P<0.05). Inhibitor could significantly reverse the improvement effect of PLT on the above indexes in AP rats (P< 0.05). CONCLUSIONS PLT inhibits inflammation and oxidative stress in AP rats, alleviates intestinal damage, and its mechanism may be related to inhibiting protein expression of Keap1 and activating Nrf2/HO-1 signaling pathway.
9.Analysis of Mechanism of Xingpi Capsules in Treatment of Functional Dyspepsia Based on Transcriptomics
Rongxin ZHU ; Mingyue HUANG ; Keyan WANG ; Xiangning LIU ; Yinglan LYU ; Gang WANG ; Fangfang RUI ; Qiong DENG ; Jianteng DONG ; Yong WANG ; Chun LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):164-172
ObjectiveTo investigate the ameliorative effect of Xingpi capsules on functional dyspepsia(FD) and the potential mechanism. MethodsSixty SPF-grade male SD neonatal rats(7 days old) were randomly divided into the normal group(n=12) and the modeling group(n=48), and the FD model was prepared by iodoacetamide gavage in the modeling group. After the model was successfully prepared, the rats in the modeling group were randomly divided into the model group, the low-dose and high-dose groups of Xingpi capsules(0.135, 0.54 g·kg-1) and the domperidone group(3 mg·kg-1), with 12 rats in each group. Rats in the normal and model groups were gavaged with distilled water, and rats in the rest of the groups were gavaged with the corresponding medicinal solution, once a day for 7 d. The general survival condition of the rats was observed, and the water intake and food intake of the rats were measured, the gastric emptying rate and the small intestinal propulsion rate were measured at the end of the treatment, the pathological damage of the rat duodenum was examined by hematoxylin-eosin(HE) staining, and the expressions of colonic tight junction protein(Occludin) and zonula occludens protein-1(ZO-1) were detected by immunofluorescence. The differentially expressed genes in the duodenal tissues of the model group and the normal group, and the high-dose group of Xingpi capsules and the model group were detected by transcriptome sequencing after the final administration, and Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were carried out. The transcriptomic results were validated by Western blot, immunofluorescence, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), and the active ingredients of Xingpi capsules were screened for molecular docking with the key targets. ResultsCompared with the normal group, the general survival condition of rats in the model group was poorer, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly reduced(P<0.05), inflammatory infiltration was seen in duodenal pathology, and the fluorescence intensities of Occludin and ZO-1 in the colon were significantly reduced(P<0.01). Compared with the model group, the general survival condition of rats in the high-dose group of Xingpi capsules improved significantly, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly increased(P<0.05), the duodenal pathology showed a decrease in inflammatory infiltration, and the fluorescence intensities of colonic Occludin and ZO-1 were significantly increased(P<0.01). Transcriptomic results showed that Xingpi capsules might exert therapeutic effects by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) through the key genes such as Slc5a1, Abhd6. The validation results showed that compared with the normal group, the phosphorylation levels of PI3K and Akt proteins, the protein expression level of interleukin(IL)-1β, and the fluorescence intensities of IL-6 and IL-1β were significantly increased in the model group(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3, Slc5a9 and other key genes were significantly increased(P<0.01). Compared with the model group, the phosphorylation levels of PI3K and Akt, the protein expression level of IL-1β and the fluorescence intensities of IL-6 and IL-1β in the high-dose group of Xingpi capsules were significantly reduced(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3 and Slc5a9 were significantly reduced(P<0.05). Weighted gene co-expression network analysis and molecular docking results showed that E-nerolidol and Z-nerolidol in Xingpi capsules were well bound to ABDH6 protein, and linarionoside A, valerosidatum and senkirkine were well bound to Slc5a1 protein. ConclusionXingpi capsules can effectively improve the general survival and gastrointestinal motility of FD rats, its specific mechanism may be related to the inhibition of PI3K/Akt signaling pathway to alleviate the low-grade inflammation of duodenum, and E-nerolidol, Z-nerolidol, linarionoside A, valerosidatum and senkirkine may be its key active ingredients.
10.Analysis of Mechanism of Xingpi Capsules in Treatment of Functional Dyspepsia Based on Transcriptomics
Rongxin ZHU ; Mingyue HUANG ; Keyan WANG ; Xiangning LIU ; Yinglan LYU ; Gang WANG ; Fangfang RUI ; Qiong DENG ; Jianteng DONG ; Yong WANG ; Chun LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):164-172
ObjectiveTo investigate the ameliorative effect of Xingpi capsules on functional dyspepsia(FD) and the potential mechanism. MethodsSixty SPF-grade male SD neonatal rats(7 days old) were randomly divided into the normal group(n=12) and the modeling group(n=48), and the FD model was prepared by iodoacetamide gavage in the modeling group. After the model was successfully prepared, the rats in the modeling group were randomly divided into the model group, the low-dose and high-dose groups of Xingpi capsules(0.135, 0.54 g·kg-1) and the domperidone group(3 mg·kg-1), with 12 rats in each group. Rats in the normal and model groups were gavaged with distilled water, and rats in the rest of the groups were gavaged with the corresponding medicinal solution, once a day for 7 d. The general survival condition of the rats was observed, and the water intake and food intake of the rats were measured, the gastric emptying rate and the small intestinal propulsion rate were measured at the end of the treatment, the pathological damage of the rat duodenum was examined by hematoxylin-eosin(HE) staining, and the expressions of colonic tight junction protein(Occludin) and zonula occludens protein-1(ZO-1) were detected by immunofluorescence. The differentially expressed genes in the duodenal tissues of the model group and the normal group, and the high-dose group of Xingpi capsules and the model group were detected by transcriptome sequencing after the final administration, and Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were carried out. The transcriptomic results were validated by Western blot, immunofluorescence, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), and the active ingredients of Xingpi capsules were screened for molecular docking with the key targets. ResultsCompared with the normal group, the general survival condition of rats in the model group was poorer, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly reduced(P<0.05), inflammatory infiltration was seen in duodenal pathology, and the fluorescence intensities of Occludin and ZO-1 in the colon were significantly reduced(P<0.01). Compared with the model group, the general survival condition of rats in the high-dose group of Xingpi capsules improved significantly, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly increased(P<0.05), the duodenal pathology showed a decrease in inflammatory infiltration, and the fluorescence intensities of colonic Occludin and ZO-1 were significantly increased(P<0.01). Transcriptomic results showed that Xingpi capsules might exert therapeutic effects by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) through the key genes such as Slc5a1, Abhd6. The validation results showed that compared with the normal group, the phosphorylation levels of PI3K and Akt proteins, the protein expression level of interleukin(IL)-1β, and the fluorescence intensities of IL-6 and IL-1β were significantly increased in the model group(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3, Slc5a9 and other key genes were significantly increased(P<0.01). Compared with the model group, the phosphorylation levels of PI3K and Akt, the protein expression level of IL-1β and the fluorescence intensities of IL-6 and IL-1β in the high-dose group of Xingpi capsules were significantly reduced(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3 and Slc5a9 were significantly reduced(P<0.05). Weighted gene co-expression network analysis and molecular docking results showed that E-nerolidol and Z-nerolidol in Xingpi capsules were well bound to ABDH6 protein, and linarionoside A, valerosidatum and senkirkine were well bound to Slc5a1 protein. ConclusionXingpi capsules can effectively improve the general survival and gastrointestinal motility of FD rats, its specific mechanism may be related to the inhibition of PI3K/Akt signaling pathway to alleviate the low-grade inflammation of duodenum, and E-nerolidol, Z-nerolidol, linarionoside A, valerosidatum and senkirkine may be its key active ingredients.

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