ObjectiveTo observe the effects of Xiaoyaosan on glucagon-like peptide-1 （GLP-1）/GLP-1 receptor （GLP-1r） and protein kinase A （PKA）/cAMP response element binding protein （CREB）/brain-derived neurotrophic factor （BDNF） signaling pathways in the hippocampal CA1 region of rats under chronic restraint stress （CRS），and to explore the mechanism of this formula to alleviate anxiety and depression-like behaviors. Methods40 specific pathogen-free male Sprague-Dawley （SD） rats were randomly divided into normal，model，Xiaoyaosan，and fluoxetine groups，with 10 rats in each group. CRS was used to induce anxiety and depression-like behaviors. The rats in the Xiaoyaosan group were gavaged with aqueous solution of traditional Chinese medicine formula granules （7.36 g·kg^-1·d^-1），while those in the fluoxetine group were gavaged with aqueous solution of fluoxetine （2 mg·kg^-1·d^-1）. Body weight was measured on days 0，7，14，and 21 of the experiment. On days 0 and 22 of the experiment，the sucrose preference test （SPT），forced swimming test （FST），and open field test （OFT） were performed. The pathological morphology of the hippocampal CA1 region was observed by Nissl staining. The relative mRNA expression of post-synaptic density protein-95 （PSD95） and synapsin （SYP） was detected by reverse transcription quantitative real-time polymerase chain reaction. Immunohistochemistry and Western blot were used to detect expression of proteins in the GLP-1/GLP-1r and PKA/CREB/BDNF pathways in the hippocampal CA1 region. ResultsAfter CRS modeling，compared with the normal group，the rats of the model group had anxiety and depression-like behavioral manifestations，neuronal damage in the hippocampal CA1 region，significantly downregulated expression of synaptic plasticity markers PSD95 and SYP genes （P<0.01），and inhibition of GLP-1/GLP-1r and PKA/CREB/BDNF signaling pathways （P<0.05，P<0.01）. Compared with the model group，the Xiaoyaosan group exhibited alleviated anxiety and depression-like behaviors，reduced neuronal damage in the hippocampal CA1 region， significantly increased expression of PSD95 and SYP genes （P<0.01），and the activation of the GLP-1/GLP-1r and PKA/CREB/BDNF signaling pathways （P<0.05，P<0.01）. ConclusionXiaoyaosan can alleviate anxiety and depression-like behaviors in CRS rats by improving synaptic plasticity in the hippocampal CA1 region. The mechanisms may be related to the activation of the GLP-1/GLP-1r pathway and its mediated PKA/CREB/BDNF signaling pathway by the formula.

ObjectiveTo investigate the characteristics and potential mechanisms of Luotong Xianrong Yin （LTXRY） in improving lung injury in rats with idiopathic pulmonary fibrosis （IPF） by regulating glycolysis. MethodsForty specific pathogen-free （SPF） Sprague-Dawley （SD） rats were randomly divided into a sham-operated group （10 mL·kg^-1）， model group （10 mL·kg^-1）， LTXRY group （15.18 g·kg^-1）， and nintedanib group （0.1 g·kg^-1）， with 10 rats in each group. The IPF rat model was established by intratracheal instillation of bleomycin. After 28 days of gavage intervention， pulmonary function was assessed. Lung pathological changes were observed by hematoxylin-eosin （HE） and Masson staining. Enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of inflammatory factors， including tumor necrosis factor-α （TNF-α）， interleukin （IL）-1β， and IL-6， in lung tissue. Chemiluminescence assays were employed to detect lactate content and lactate dehydrogenase activity in lung tissue. Western blot was used to measure the protein expression of transforming growth factor-β₁ （TGF-β₁）， CollagenⅠ and CollagenⅢ to evaluate collagen deposition， as well as hexokinase 2 （HK2）， pyruvate kinase M2 （PKM2）， and 6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase 3 （PFKFB3） to assess glycolysis levels. Network pharmacology was applied to analyze the potential targets and signaling pathways of LTXRY in IPF， and molecular docking was conducted to evaluate the binding energy between active components and potential targets. Western blot was further used to detect the expression of target- and pathway-related proteins. ResultsCompared with the sham-operated group， rats in the model group showed significantly increased main airway resistance （Rn） and respiratory system resistance （Rrs）， and significantly decreased respiratory system compliance （Crs）. Inflammatory infiltration and collagen deposition were observed in lung tissue， with significantly increased levels of TNF-α， IL-1β， and IL-6， as well as elevated protein expression of TGF-β₁， CollagenⅠ and CollagenⅢ. Lactate content， lactate dehydrogenase activity， and the protein expression of HK2， PKM2， and PFKFB3 in lung tissue were significantly increased. Network pharmacology analysis indicated that signal transducer and activator of transcription 3 （STAT3） was a key target of LTXRY in IPF， and hypoxia-inducible factor-1 （HIF-1） was a critical signaling pathway. The expression levels of phosphorylated STAT3 （p-STAT3） and HIF-1α in lung tissue were significantly higher than those in the sham-operated group. Compared with the model group， rats in the LTXRY group showed significantly decreased Rn and Rrs and significantly increased Crs. Lung inflammatory infiltration and collagen deposition were markedly alleviated， with significantly reduced levels of TNF-α， IL-1β， and IL-6， and decreased protein expression of TGF-β₁， CollagenⅠ and CollagenⅢ. Lactate content， lactate dehydrogenase activity， and the protein expression of HK2， PKM2， and PFKFB3 were significantly decreased， accompanied by markedly reduced expression of p-STAT3 and HIF-1α. ConclusionLTXRY alleviates lung tissue injury in IPF rats by regulating glycolysis mediated by the STAT3/HIF-1α signaling pathway.

ObjectiveTo elucidate the correlation between alterations in digestion and absorption functions and hepatic deficiency states in aging rats based on the R-spondin1/Wnt3a signaling pathway， and reveal the intervention mechanism of Bugansan. MethodsForty-eight SPF-grade male SD rats were randomly assigned to six groups： blank control， model， low-， medium-， and high-dose （7.03， 14.06， 28.12 g·kg^-1， respectively） Bugansan， and vitamin E （suspension， 27 mg·kg^-1）， with 8 rats in each group. The rat model of aging was established by intraperitoneal injection of D-galactose （400 mg·kg^-1）， while the blank control group was injected with normal saline. Since the day of modeling， rats in intervention groups received corresponding agents by gavage， and those in blank control and model groups received an equal volume of normal saline （10 mL·kg^-1）. General biological features such as fur color， activity， body mass， water intake， and food intake were observed. Meanwhile， the content of malondialdehyde （MDA） and the activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） in the serum were measured to assess aging. Grip strength and the content of total bile acids （TBA） and the activity of α-amylase （AMY） in the serum were measured to evaluate hepatic deficiency states. The activity of β-galactosidase （β-gal） in the duodenum was measured to evaluate intestinal senescence. The levels of glucagon-like peptide-1 （GLP-1）， vasoactive intestinal peptide （VIP）， and D-xylose in the serum were determined to assess digestion and absorption functions of the small intestine. Hematoxylin-eosin staining was conducted to observe pathological changes of the duodenum to assess the small intestine damage. Immunohistochemical staining was employed to visualize the expression of B-cell-specific Moloney murine leukemia virus integration site 1 （Bmi1） and leucine-rich repeat-containing G protein-coupled receptor 5 （Lgr5） in the duodenal tissue. Moreover， Real-time quantitative polymerase chain reaction （Real-time PCR） was utilized to quantify the mRNA levels of Ki67， Bmi1， and Lgr5 to assess proliferation and regeneration of the small intestine. Additionally， the mRNA levels of R-spondin1， Wnt3a， β-catenin， and glycogen synthase kinase-3β （GSK-3β） and the protein levels of R-spondin1， Wnt3a， β-catenin， and phosphorylated GSK-3β （p-GSK-3β） in the duodenum were determined by Real-time PCR and Western blot， respectively， to analyze the mechanisms of intestinal digestion and absorption dysfunction in aging rats and the regulatory characteristics of Bugansan. ResultsCompared with blank control group， the model group showed decreases in body mass， water intake， food intake， grip strength， activities of SOD， GSH-Px， and AMY in the serum and content of GLP-1， VIP and D-xylose in the serum （P<0.05）， increases in the content of MDA and TBA in the serum and β-gal activity in the duodenum （P<0.05）， reductions in villus length， villus width， crypt depth， and villi/crypt （V/C） value， down-regulated mRNA and protein levels of Ki67， Lgr5， Bmi1， R-spondin1， Wnt3a， β-catenin， and up-regulated level of GSK-3β， phosphorylation （p）-GSK-3β （P<0.05）. Compared with the model group， Bugansan increased the body mass， water intake， food intake， grip strength， and activities of SOD， GSH-Px， and AMY and levels of GLP-1， VIP and D-xylose in the serum （P<0.05）， while decreasing the content of MDA and TBA in the serum and β-gal activity in the duodenum （P<0.05）. Furthermore， Bugansan increased the villus length， villus width， crypt depth， and V/C value， up-regulated the mRNA and protein levels of Ki67， Lgr5， Bmi1， R-spondin1， Wnt3a， β-catenin， and down-regulated the level of GSK-3β and p-GSK-3β （P<0.05）. ConclusionAging rats exhibit obvious impairments in digestion and absorption functions， accompanied by a state of hepatic deficiency. The traditional Chinese medicine approach of tonifying liver Qi effectively ameliorates aging-related changes by modulating the R-spondin1/Wnt3a signaling pathway to mitigate intestinal senescence and enhance digestion and absorption functions， ultimately contributing to the delay of aging.

Objective To investigate the correlation between PPARγ and female interstitial cystitis/bladder pain syndrome (IC/BPS) and to establish a predictive model. Methods Clinical data were collected from 89 female IC/BPS patients (observational group) admitted to the hospital from June 2022 to December 2023,and 90 healthy female volunteers undergoing physical examinations during the same period (control group). Plasma levels of inflammatory factors,total antioxidant capacity (TAC),total glutathione (GSH),malondialdehyde (MDA),and PPARγ levels were measured. Significant clinical features were identified using LASSO regression and fitted into a multivariate logistic regression model. The diagnostic efficacy was assessed through receiver operat-ing characteristic (ROC) curves. Results Compared to the control group,the observation group exhibited signifi-cantly elevated age,BMI,NLR,absolute neutrophil count,IFN-α,IL-1β,IL-6,IL-8,TNF-α and CD3+CD4+T expression levels,while absolute lymphocyte count,IL-10,TAC,GSH and plasma PPARγ expression levels were significantly decreased (all P<0.05). LASSO regression identified 8 variables,including NLR,IFN-α,absolute neutrophil count,IL-1β,IL-6,TNF-α,CD3+CD4+T and PPARγ,which were incorporated into the predictive model. Multivariate binary logistic regression analysis revealed that elevated levels of IL-1β,TNF-α and CD3+CD4+T cells,along with reduced PPARγ levels,were independent risk factors for IC/BPS. ROC curve analy-sis indicated that the diagnostic efficacy of the combined PPARγ and clinical parameters (age,IL-1β,TNF-α and CD3+CD4+T) (AUC=0.901) was superior to PPARγ alone (AUC=0.839). Conclusion Plasma PPARγ levels are significantly reduced in female IC/BPS patients and serve as a potential diagnostic marker,with combined clinical parameters enhancing diagnostic accuracy.

AIM:To investigate the mechanism of Luotong-Xianrong Yin(LTXRY)in regulating endoplasmic reticulum stress(ERS)to improve lung function of rats with idiopathic pulmonary fibrosis(IPF).METHODS:Forty-eight SPF male SD rats were randomly divided into sham group,model group,LTXRY group and nintedanib group,with 12 rats in each group.The IPF rat model was prepared by intratracheal infusion of bleomycin.The rats in LTXRY group were gavaged with aqueous solution of Chinese medicine formula granules,while the rats in nintedanib group were gavaged with aqueous solution of nintedanib.Lung function was detected by awake and unrestrained animal lung function test sys-tem.The pathomorphologic change in lung tissue was observed by HE and Masson staining.The expression of α-smooth muscle actin(α-SMA)was observed by immunohistochemical(IHC)staining,and the expression of vimentin was detect-ed by Western blot.The expressions of Bax,Bcl2,cleaved caspase-3,and PERK/eIF2α signal pathway related proteins were detected by Western blot.RESULTS:Abnormal lung function and fibrotic-like changes in lung tissue were present in rats in the IPF state.After the intervention of LTXRY,the lung function of IPF rats was significantly improved(P<0.01),the degree of lung tissue damage was significantly reduced(P<0.01),the expressions of apoptosis-related pro-teins,Bax and cleaved caspase-3,were significantly decreased(P<0.05 or P<0.01),while the expression of Bcl2 was significantly increased(P<0.05).Moreover,the ERS signal pathway,PERK/eIF2α,was significantly inhibited(P<0.05 or P<0.01).CONCLUSION:LTXRY could improve lung function in IPF rats by regulating cell apoptosis,and the mechanism may be related to inhibit endoplasmic reticulum stress.

Objective To investigate the correlation between PPARγ and female interstitial cystitis/bladder pain syndrome (IC/BPS) and to establish a predictive model. Methods Clinical data were collected from 89 female IC/BPS patients (observational group) admitted to the hospital from June 2022 to December 2023,and 90 healthy female volunteers undergoing physical examinations during the same period (control group). Plasma levels of inflammatory factors,total antioxidant capacity (TAC),total glutathione (GSH),malondialdehyde (MDA),and PPARγ levels were measured. Significant clinical features were identified using LASSO regression and fitted into a multivariate logistic regression model. The diagnostic efficacy was assessed through receiver operat-ing characteristic (ROC) curves. Results Compared to the control group,the observation group exhibited signifi-cantly elevated age,BMI,NLR,absolute neutrophil count,IFN-α,IL-1β,IL-6,IL-8,TNF-α and CD3+CD4+T expression levels,while absolute lymphocyte count,IL-10,TAC,GSH and plasma PPARγ expression levels were significantly decreased (all P<0.05). LASSO regression identified 8 variables,including NLR,IFN-α,absolute neutrophil count,IL-1β,IL-6,TNF-α,CD3+CD4+T and PPARγ,which were incorporated into the predictive model. Multivariate binary logistic regression analysis revealed that elevated levels of IL-1β,TNF-α and CD3+CD4+T cells,along with reduced PPARγ levels,were independent risk factors for IC/BPS. ROC curve analy-sis indicated that the diagnostic efficacy of the combined PPARγ and clinical parameters (age,IL-1β,TNF-α and CD3+CD4+T) (AUC=0.901) was superior to PPARγ alone (AUC=0.839). Conclusion Plasma PPARγ levels are significantly reduced in female IC/BPS patients and serve as a potential diagnostic marker,with combined clinical parameters enhancing diagnostic accuracy.

AIM:To investigate the mechanism of Luotong-Xianrong Yin(LTXRY)in regulating endoplasmic reticulum stress(ERS)to improve lung function of rats with idiopathic pulmonary fibrosis(IPF).METHODS:Forty-eight SPF male SD rats were randomly divided into sham group,model group,LTXRY group and nintedanib group,with 12 rats in each group.The IPF rat model was prepared by intratracheal infusion of bleomycin.The rats in LTXRY group were gavaged with aqueous solution of Chinese medicine formula granules,while the rats in nintedanib group were gavaged with aqueous solution of nintedanib.Lung function was detected by awake and unrestrained animal lung function test sys-tem.The pathomorphologic change in lung tissue was observed by HE and Masson staining.The expression of α-smooth muscle actin(α-SMA)was observed by immunohistochemical(IHC)staining,and the expression of vimentin was detect-ed by Western blot.The expressions of Bax,Bcl2,cleaved caspase-3,and PERK/eIF2α signal pathway related proteins were detected by Western blot.RESULTS:Abnormal lung function and fibrotic-like changes in lung tissue were present in rats in the IPF state.After the intervention of LTXRY,the lung function of IPF rats was significantly improved(P<0.01),the degree of lung tissue damage was significantly reduced(P<0.01),the expressions of apoptosis-related pro-teins,Bax and cleaved caspase-3,were significantly decreased(P<0.05 or P<0.01),while the expression of Bcl2 was significantly increased(P<0.05).Moreover,the ERS signal pathway,PERK/eIF2α,was significantly inhibited(P<0.05 or P<0.01).CONCLUSION:LTXRY could improve lung function in IPF rats by regulating cell apoptosis,and the mechanism may be related to inhibit endoplasmic reticulum stress.

Objective:To explore the mechanism of Yupingfeng Powder on patients with chronic obstructive pulmonary disease (COPD) through UHPLC-QE-MS combined with network pharmacology and molecular docking technology.Methods:UHPLC-QE-MS technology was used to detect the chemical composition of Yupingfeng Powder, and its targets were obtained using ChEMB and TCMIO databases; COPD treatment targets were obtained from TTD, GeneCards, SymMap, and DisGeNET databases, and the intersection of the two was taken. GO enrichment analysis and KEGG pathway analysis were performed on intersection targets. Top 20 metabolites with the most matching targets were selected, their corresponding targets with the top 20 KEGG pathway targets were merged, and potential targets for the treatment of COPD with Yupingfeng Powder were obtained. A target protein-target protein interaction network (PPI) was constructed, key targets of Yupingfeng Powder for the treatment of COPD were screened, and molecular docking was used for validation.Results:Under positive and negative ion modes, a total of 73 active components were identified in Yupingfeng Powder. The top three ranked in terms of degree were genistein, apigenin, and kaempferol. 449 targets of 73 active components and 3 455 disease targets were obtained from the database. The intersection of the two was obtained, resulting in 294 common targets. A total of 69 pathways were identified through KEGG pathway analysis, involved in the pathogenesis of cancer, non-small cell lung cancer signaling pathway, VEGF signaling pathway, T cell receptor (TCR) signaling pathway, arachidonic acid metabolism, Toll like receptor signaling pathway, etc. GO enrichment analysis obtained 2 647 biological processes, 126 cellular components, and 289 molecular functions, involving reactions to nutritional levels, extracellular stimuli, oxidative stress, etc. The top 20 KEGG pathways and the top 20 metabolites with the most matching targets correspond to a total of 281 targets. The key targets were TP53, STAT3, c-Jun, AKT1, and ESR1. The molecular docking results showed that genistein, apigenin, and kaempferol bound well with core targets TP53, STAT3, and c-Jun.Conclusion:Yupingfeng Powder may affect the levels of inflammation, immune regulation, and oxidative stress in COPD through multiple components, targets, and pathways, thereby affecting the progression of COPD.

ObjectiveTo observe the effect of Shengxiantang （SXT） on cell senescence mediated by wingless/integrated （Wnt）3a/β-catenin pathway in rats with idiopathic pulmonary fibrosis （IPF） and reveal the possible mechanism in improving lung function of IPF rats. MethodA total of 32 SPF level SD rats were randomly divided into sham group， model group， pirfenidone group， and SXT group. The IPF rat model was established by intratracheal instillation of bleomycin （0.005 g·kg^-1）. The following day after surgery， rats in the SXT group were given the aqueous solution of SXT granules （0.78 g·kg^-1）， and the pirfenidone group was given pirfenidone suspension （0.05 g·kg^-1）. The other groups were given deionized water （10 mL·kg^-1） for 28 consecutive days. Lung tissue was collected after the lung function was measured. The pathological changes of the lung tissue were observed by hematoxylin-eosin （HE） and Masson staining， and then the Szapiel score and Ashcroft score were performed. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect telomere length. Western blot was applied to detect the expressions of epithelial-mesenchymal transformation （EMT） markers ［α-smooth muscle actin （α-SMA） and E-cadherin］， telomere reverse transcriptase （TRET）， aging-related proteins （p53 and p21）， senescence-associated secretory phenotype ［interleukin-6 （IL-6） and matrix metalloproteinase-1 （MMP-1）］， and key proteins of Wnt signaling pathway ［Wnt3a， glycogen synthase kinase-3β （GSK-3β）， β-catenin， Cyclin D₁， and c-Myc］. ResultCompared with those in the Sham group， peak expiratory flow （PEF） and minute ventilation volume （MV） in the model group were significantly decreased （P<0.01）， and the frequency of respiratory （f） was significantly increased （P<0.01）. The Szapiel score， Ashcroft score， and protein expression of α-SMA， p53， p21， IL-6， MMP-1， Wnt3a， GSK3β， β-catenin， Cyclin D₁， and c-Myc were increased （P<0.01）. The expressions of E-cadherin and TERT， as well as telomere length were significantly decreased （P<0.01）. Compared with those in the model group， PEF and MV in the SXT group were significantly increased （P<0.01）， while f was significantly decreased （P<0.01）. The Szapiel score， Ashcroft score， and protein expression of α-SMA， p53， p21， IL-6， MMP-1， Wnt3a， GSK3β， β-catenin， Cyclin D₁， and c-Myc were significantly decreased （P<0.05， P<0.01）. Nevertheless， the expression of E-cadherin and TERT， as well as telomere length were significantly increased （P<0.01）. ConclusionSXT presents a significant protective effect on lung function in IPF rats， and the prescription may act on the Wnt3a/β-catenin signaling pathway to regulate cell senescence induced by TERT to inhibit EMT.

ObjectiveTo investigate the effect of shikosin （SHI） on psoriasis （PSO） and explore the underlying mechanism via the cyclic guanosine monophosphate-adenosine monophosphate synthase （cGAS）/stimulator of interferon genes （STING） signaling pathway. MethodHaCaT cells were classified into normal culture（Control）， a mixture of five proinflammatory cytokines（M5）， low-， medium-， and high-dose SHI （L-SHI， M-SHI， and H-SHI， respectively）， and SHI+ADU-S100 groups. The cells in the M5 group were stimulated with 10 μg·L^-1 interleukin （IL）-1α， IL-17， IL-22， tumor necrosis factor （TNF）-α， and oncostatin M （OSM） for 48 h. The cells in the L-SHI， M-SHI， and H-SHI groups were treated with 0.1， 1， 10 μmol·L^-1 SHI， respectively， on the basis of the treatment in the M5 group. The cells in the SHI+ADU-S100 group were treated with 10 μmol·L^-1 STING activator ADU-S100 on the basis of the treatment in the H-SHI group. The methyl thiazolyl tetrazolium （MTT） assay and colony formation assay were employed to examine the effect of SHI on the proliferation of HaCaT cells. The wound healing assay was employed to examine the effect of SHI on the migration of HaCaT cells. Flow cytometry was employed to detect the effect of SHI on the apoptosis of HaCaT cells. Enzyme-linked immunosorbent assay was employed to measure the levels of IL-1β， IL-6， IL-15， IL-23， and interferon-γ （IFN-γ） in HaCaT cells. Western blot was employed to determine the protein levels of cGAS and STING in HaCaT cells. ResultCompared with Control group， the M5 group showed decreased survival rate， colony formation， and would healing rate of HaCaT cells， increased apoptosis rate， elevated levels of IL-1β， IL-6， IL-15， IL-23， and IFN-γ， and up-regulated protein levels of cGAS and STING （P<0.01）. Compared with the M5 group， the L-SHI， M-SHI， and H-SHI groups showed increased survival rate， cell colony formation， and wound healing rate， decreased apoptosis rate， lowered levels of IL-1β， IL-6， IL-15， IL-23， and IFN-γ， and down-regulated protein levels of cGAS and STING （P<0.01）. Compared with the H-SHI group， the SHI+ADU-S100 group showed decreased survival rate， cell colony formation， and wound healing rate， increased apoptosis rate， risen levels of IL-1β， IL-6， IL-15， IL-23， and IFN-γ， and up-regulated protein levels of cGAS and STING （P<0.01）. ConclusionSHI can inhibit the inflammation in the cell model of PSO by inhibiting the cGAS/STING signaling pathway.