Objective To observe the effects of Yifei Jianpi Prescription on airway mucus hypersecretion and protein expressions of EGFR/PKC/NF-κB pathway in lipopolysaccharide(LPS)-induced acute lung injury(ALI)model rats;To explore its mechanism in the treatment of ALI.Methods Ten of 60 SPF SD rats were randomly selected as blank group,and the other rats were intratracheal instilled with LPS to establish ALI model.The model rats were randomly divided into model group,dexamethasone group and Yifei Jianpi Prescription high-,medium-and low-dosage groups,with 8 rats in each group.Each treatment group was given corresponding drug solution by gavage,and the blank group and model group were given equal volume of normal saline by gavage,once a day for 14 days.The pulmonary functions of rats were measured[peak expiratory flow(PEF),tidal volume(TV),expiratory volume(EV),50%expiratory flow rate(EF50)],HE staining was used to observe the morphology of lung tissue,AB-PAS staining was used to evaluate the proliferation and mucus secretion of goblet cells,the expressions of epidermal growth factor receptor(EGFR),protein kinase C(PKC),nuclear factor-κB(NF-κB)p65 and MUC5AC in lung tissue were detected by immunofluorescence staining,the mRNA expressions of EGFR and MUC5AC in lung tissue were detected by fluorescent quantitative PCR,and the content of MUC5AC in lung tissue was detected by ELISA.Results Compared with the blank group,PEF,TV,EV and EF50 of the model group rats significantly decreased(P<0.01);the bronchial wall was significantly thickened,the lumen narrowed,pulmonary interstitial edema and hyperemia,the thickness of alveolar wall increased,accompanied by a large number of inflammatory cells infiltration,and the lung tissue injury score increased significantly(P<0.01);goblet cells proliferated significantly,mucus secretion increased significantly(P<0.01);the protein expressions of EGFR,PKC,NF-κB p65,MUC5AC and mRNA expressions of EGFR and MUC5AC in lung tissue increased significantly(P<0.01),and the content of MUC5AC in lung tissue increased significantly(P<0.01).Compared with the model group,PEF,TV,EV and EF50 in dexamethasone group and Yifei Jianpi Prescription each dosage groups increased in varying degrees;the pathological injury of lung tissue was alleviated to varying degrees,the score of lung tissue injury was reduced;the proliferation of goblet cells was reduced,and the secretion of mucus was reduced,the expressions of EGFR,PKC,NF-κB p65,MUC5AC protein and EGFR,MUC5AC mRNA in lung tissue decreased,and the content of MUC5AC in lung tissue decreased.There was statistical significance in dexamethasone group and Yifei Jianpi Prescription high-and medium-dosage groups(P<0.01).Conclusion Yifei Jianpi Prescription can inhibit the hypersecretion of airway mucus and the high expression of EGFR/PKC/NF-κB pathway protein in rats with ALI induced by LPS.

ObjectiveTo observe the effect of Yifei Jianpi prescription on the of signal transducer and activator of transcription protein 1 （STAT1）/interferon regulatory factor 3 （IRF3） signaling pathway in a pneumonia model induced by lipopolysaccharide （LPS） and to explore the mechanism of Yifei Jianpi prescription in improving lung immune and inflammatory responses. MethodsSixty male SPF SD rats were used in this study. Ten rats were randomly assigned to the normal control group， and the remaining 50 were instilled with LPS in the trachea to establish a pneumonia model. After successful modeling， the rats were randomly divided into the model group， dexamethasone group （0.5 mg·kg^-1）， and Yifei Jianpi prescription high-dose （12 mg·kg^-1）， medium-dose （6 mg·kg^-1）， and low-dose （3 mg·kg^-1） groups， with 10 rats in each group. Treatment was administered once daily， and the normal control and model groups received the same volume of normal saline. After 14 days， flow cytometry was used to detect the classification of whole blood lymphocytes. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum levels of immunoglobulin G （IgG）， immunoglobulin A （IgA）， immunoglobulin M （IgM）， and the content of tumor necrosis factor-α （TNF-α）， interleukin-8 （IL-8）， interleukin-6 （IL-6）， and interleukin-10 （IL-10） in alveolar lavage fluid （BALF）. Hematoxylin-eosin （HE） staining was used to observe lung tissue pathology and score the damage. Thymus weight， spleen weight， and wet-to-dry weight ratio （W/D） were recorded. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of STAT1， IRF3， IL-6， and interferon-alpha （IFN-α） in lung tissues， while Western blot was performed to assess the protein expression of STAT1， IRF3， IL-6， and IFN-α. ResultsCompared with the normal control group， the model group showed significantly increased proportion of B lymphocytes in peripheral blood， decreased proportions of NK cells and CD4⁺/CD8⁺ （P<0.05， P<0.01）， decreased serum levels of IgG and IgA， significantly increased IgM levels （P<0.01）， significantly elevated content of TNF-α， IL-6， and IL-8 in BALF， and significantly decreased IL-10 levels （P<0.01）. Lung tissue damage was evident， with significant increases in thymus and spleen weights and a higher W/D ratio （P<0.01）. The mRNA and protein expression of STAT1， IRF3， IFN-α， and IL-6 in lung tissues was significantly upregulated （P<0.05，P<0.01）. Compared with the model group， the Yifei Jianpi prescription groups showed significantly reduced proportions of B lymphocytes in peripheral blood， increased proportions of NK cells and CD4⁺/CD8⁺ ratios （P<0.05， P<0.01）， significantly increased serum levels of IgG and IgA， significantly decreased IgM levels （P<0.05， P<0.01）， significantly reduced levels of TNF-α， IL-6， and IL-8 in BALF， and significantly increased IL-10 levels （P<0.01）. Lung tissue damage was alleviated， thymus and spleen weights were significantly reduced， and the W/D ratio was markedly decreased （P<0.01）. The mRNA and protein expression of STAT1， IRF3， IFN-α， and IL-6 in lung tissues was significantly downregulated （P<0.05， P<0.01）. ConclusionYifei Jianpi prescription can alleviate lung tissue damage and improve immune and inflammatory responses in LPS-induced pneumonia rats. The mechanism may be related to the inhibition of STAT1/IRF3 signaling pathway activation.

Objective To observe the synergistic effect of Shengxian Huaxian Prescription and its disassembled prescription on pulmonary fibrosis;To explore whether its mechanism is related to regulating the STAT6/PPAR-y pathway to promote polarization of M2 type macrophages towards M1 type.Methods Ten SD rats were randomly selected from 70 rats as blank group,and the remaining rats were re-established pulmonary fibrosis model by intratracheal infusion of bleomycin.After modeling,the rats were divided into model group,positive group,Shengxian Huaxian Prescription group,Shengxian group,Tongluo group and Bushen group,with 10 rats in each group.Shengxian Huaxian Prescription group,Shengxian group,Tongluo group and Bushen group were given 12.60,7.65,3.60 and 2.25 g/kg of corresponding TCM solution,respectively;the positive group was given 0.12 g/kg of pirfenidone suspension;the blank group and the model group were given equal volume of normal saline,once a day,for consecutive 28 days.The lung function of rats was detected,the contents of IL-6 and TGF-β1 in serum were detected by ELISA,the pathological changes in lung tissue were observed by Masson staining,the expression of CD68,iNOS and CD206,Arg-1 in lung tissue were detected by immunofluorescence,the expression of SOCS1,SOCS3,STAT6,p-STAT6 and PPAR-γ in lung tissue were detected by Western blot.Results Compared with the blank group,the PEF,PIF and EF50 in model group rats significantly decreased,and the contents of serum IL-6 and TGF-β1 significantly increased,Masson staining showed a large amount of collagen fiber deposition,Ashcroft score significantly increased,CD206,Arg-1,STAT6,p-STAT6,PPAR-y protein expression significantly increased(P＜0.01),the expressions of SOCS1 and SOCS3 protein significantly decreased(P＜0.01),while the expression of CD68 and iNOS were not significantly changed(P＞0.05).Compared with the model group,the PEF,PIF and EF50 in all administration groups significantly increased,and the contents of serum IL-6 and TGF-β1 significantly decreased,collagen fiber deposition in lung tissue were decreased to varying degree,Ashcroft score significantly decreased,the expression of CD206,Arg-1,STAT6,p-STAT6 and PPAR-γ protein significantly decreased(P＜0.01),the expressions of CD68,iNOS,SOCS1 and SOCS3 protein significantly increased(P＜0.05).The above indicators showed the most significant changes in Shengxian Huaxian Prescription group,followed by Shengxian group(P＜0.01,P＜0.05).Conclusion Both Shengxian Huaxian Prescription and its disassembled prescription have anti pulmonary fibrosis effects,and their mechanism may related to regulating the STAT6/PPAR-y pathway and promoting polarization of M2 type macrophages towards M1 type.Shengxian Huaxian Prescription group has the best effect,while Shengxian group play an important role in the prescription,and the compatibility between each group has a synergistic effect.

Objective:To investigate impacts of paeoniflorin on inflammation and nuclear transcription factor-κB(NF-κB)/nucleotide-binding oligomeric domain-like receptor protein 3(NLRP3)signal pathway in knee osteoarthritis(KOA)model rats.Meth-ods:SD rats were randomly grouped into model group,glucosamine group,paeoniflorin low-dose group,paeoniflorin high-dose group,paeoniflorin high-dose+phorbol ester(PMA)(NF-κB activator)group,with 10 rats in each group,another 10 rats were regarded as control group,and only knee joint capsule was cut,after treatment with glucosamine,paeoniflorin and PMA,joint pain symptoms of rats were detected by mechanical stimulation method and thermal radiation method;knee joint width,joint swelling and synovial thick-ness were measured;HE staining was applied to detect joint tissue structure and morphology of rats in each group;levels of inflamma-tory factors IL-1β,monocyte chemoattractant protein 1(MCP-1)and IL-9 in joint fluid and serum of rats were detected by ELISA;and Western blot was applied to detect expressions of NF-κB/NLRP3 signal pathway related proteins in rat joint tissue.Results:Com-pared with control group,joint tissue structure of model group was significantly damaged,mechanical pain threshold and thermal sen-sitivity pain threshold were significantly lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05).Compared with model group,joint tissue damage of rats in glucosamine group,paeoniflorin low-dose group and paeoniflorin high-dose group was reduced,mechanical pain threshold and thermal sensitivity pain threshold were higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expressions of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were lower(P<0.05);joint tissue injury of rats in paeoniflorin high-dose group was further reduced compared with paeoniflorin low-dose group,mechanical pain threshold and thermal sensitivity pain threshold were further higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were further lower(P<0.05).Compared with paeoniflorin high-dose group,joint tissue damage of rats in paeoniflorin high-dose+PMA group was increased,mechanical pain threshold and thermal sensitivity pain threshold were lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05);there was no significant change in all indexes of rats in glucosamine group(P>0.05).Conclusion:Paeoniflorin can play an anti-inflammatory role by down-regulating the NF-κB/NLRP3 signal pathway,thus alleviating joint injury and joint pain symptoms of KOA rats.

Objective:To study the effect of anthocyanins(C3G)on cuproptosis in chronic epileptic rats.Methods:Chronic epileptic rat model was induced by pentatetrazol(PTZ),and 90 SD rats were randomly divided into control group,PTZ group,elesclomol(ELC)group,tetrathiomolybdate(TTM)group,C3G group and ELC+C3G group.The grade,latency and frequency of seizures were recorded in each group.electroencephalogram(EEG)was used to detect abnormal electrical discharge in the brain.The action potential of hippocampal neurons was measured by patch-clamp technique.The contents of glutathione(GSH)and cuprous ions(Cu+)in hippocampus were determined by kit.Neuron damage in hippocampus was evaluated by Nissl staining.The expression of ferredoxin1(FDX1)and lipoic acid synthase(LIAS)in hippocampus was analyzed by immunohistochemistry and Western blot.Results:Compared with the control group,the rats in the PTZ group exhibited epileptic-like seizures,suggesting that the modeling was successful.Com-pared with other epileptic groups,the ELC group showed increased seizure grade,more abnormal discharges,shortened latency period,enhanced neuronal excitability,decreased Nissl particles,elevated Cu+levels,decreased GSH levels,and increased expressions of FDX1 and LIAS.The reverse was observed in C3G group(P＜0.05).Neuron damage in ELC+C3G group was less severe than that in ELC group,but more than that in PTZ group(P＜0.05).Neuron dam-age in TTM group was less severe than that in PTZ group,but more severe than that in C3G group(P＜0.05).Conclusion:cuproptosis exists in hippocampus of rats with chronic epilepsy,and the C3G can significantly inhibit cu-proptosis and alleviate the occurrence and development of chronic epilepsy.

Objective:To study the effect of anthocyanins(C3G)on cuproptosis in chronic epileptic rats.Methods:Chronic epileptic rat model was induced by pentatetrazol(PTZ),and 90 SD rats were randomly divided into control group,PTZ group,elesclomol(ELC)group,tetrathiomolybdate(TTM)group,C3G group and ELC+C3G group.The grade,latency and frequency of seizures were recorded in each group.electroencephalogram(EEG)was used to detect abnormal electrical discharge in the brain.The action potential of hippocampal neurons was measured by patch-clamp technique.The contents of glutathione(GSH)and cuprous ions(Cu+)in hippocampus were determined by kit.Neuron damage in hippocampus was evaluated by Nissl staining.The expression of ferredoxin1(FDX1)and lipoic acid synthase(LIAS)in hippocampus was analyzed by immunohistochemistry and Western blot.Results:Compared with the control group,the rats in the PTZ group exhibited epileptic-like seizures,suggesting that the modeling was successful.Com-pared with other epileptic groups,the ELC group showed increased seizure grade,more abnormal discharges,shortened latency period,enhanced neuronal excitability,decreased Nissl particles,elevated Cu+levels,decreased GSH levels,and increased expressions of FDX1 and LIAS.The reverse was observed in C3G group(P＜0.05).Neuron damage in ELC+C3G group was less severe than that in ELC group,but more than that in PTZ group(P＜0.05).Neuron dam-age in TTM group was less severe than that in PTZ group,but more severe than that in C3G group(P＜0.05).Conclusion:cuproptosis exists in hippocampus of rats with chronic epilepsy,and the C3G can significantly inhibit cu-proptosis and alleviate the occurrence and development of chronic epilepsy.

Objective:To investigate impacts of paeoniflorin on inflammation and nuclear transcription factor-κB(NF-κB)/nucleotide-binding oligomeric domain-like receptor protein 3(NLRP3)signal pathway in knee osteoarthritis(KOA)model rats.Meth-ods:SD rats were randomly grouped into model group,glucosamine group,paeoniflorin low-dose group,paeoniflorin high-dose group,paeoniflorin high-dose+phorbol ester(PMA)(NF-κB activator)group,with 10 rats in each group,another 10 rats were regarded as control group,and only knee joint capsule was cut,after treatment with glucosamine,paeoniflorin and PMA,joint pain symptoms of rats were detected by mechanical stimulation method and thermal radiation method;knee joint width,joint swelling and synovial thick-ness were measured;HE staining was applied to detect joint tissue structure and morphology of rats in each group;levels of inflamma-tory factors IL-1β,monocyte chemoattractant protein 1(MCP-1)and IL-9 in joint fluid and serum of rats were detected by ELISA;and Western blot was applied to detect expressions of NF-κB/NLRP3 signal pathway related proteins in rat joint tissue.Results:Com-pared with control group,joint tissue structure of model group was significantly damaged,mechanical pain threshold and thermal sen-sitivity pain threshold were significantly lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05).Compared with model group,joint tissue damage of rats in glucosamine group,paeoniflorin low-dose group and paeoniflorin high-dose group was reduced,mechanical pain threshold and thermal sensitivity pain threshold were higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expressions of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were lower(P<0.05);joint tissue injury of rats in paeoniflorin high-dose group was further reduced compared with paeoniflorin low-dose group,mechanical pain threshold and thermal sensitivity pain threshold were further higher(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were further lower(P<0.05).Compared with paeoniflorin high-dose group,joint tissue damage of rats in paeoniflorin high-dose+PMA group was increased,mechanical pain threshold and thermal sensitivity pain threshold were lower(P<0.05),knee joint width,joint swelling and synovial thickness,levels of IL-1β,MCP-1 and IL-9 in joint fluid and serum,and expression of p-NF-κB p65/NF-κB p65 and NLRP3 proteins in joint tissue were higher(P<0.05);there was no significant change in all indexes of rats in glucosamine group(P>0.05).Conclusion:Paeoniflorin can play an anti-inflammatory role by down-regulating the NF-κB/NLRP3 signal pathway,thus alleviating joint injury and joint pain symptoms of KOA rats.

Objective To observe the effects of Yifei Jianpi Prescription on airway mucus hypersecretion and protein expressions of EGFR/PKC/NF-κB pathway in lipopolysaccharide(LPS)-induced acute lung injury(ALI)model rats;To explore its mechanism in the treatment of ALI.Methods Ten of 60 SPF SD rats were randomly selected as blank group,and the other rats were intratracheal instilled with LPS to establish ALI model.The model rats were randomly divided into model group,dexamethasone group and Yifei Jianpi Prescription high-,medium-and low-dosage groups,with 8 rats in each group.Each treatment group was given corresponding drug solution by gavage,and the blank group and model group were given equal volume of normal saline by gavage,once a day for 14 days.The pulmonary functions of rats were measured[peak expiratory flow(PEF),tidal volume(TV),expiratory volume(EV),50%expiratory flow rate(EF50)],HE staining was used to observe the morphology of lung tissue,AB-PAS staining was used to evaluate the proliferation and mucus secretion of goblet cells,the expressions of epidermal growth factor receptor(EGFR),protein kinase C(PKC),nuclear factor-κB(NF-κB)p65 and MUC5AC in lung tissue were detected by immunofluorescence staining,the mRNA expressions of EGFR and MUC5AC in lung tissue were detected by fluorescent quantitative PCR,and the content of MUC5AC in lung tissue was detected by ELISA.Results Compared with the blank group,PEF,TV,EV and EF50 of the model group rats significantly decreased(P<0.01);the bronchial wall was significantly thickened,the lumen narrowed,pulmonary interstitial edema and hyperemia,the thickness of alveolar wall increased,accompanied by a large number of inflammatory cells infiltration,and the lung tissue injury score increased significantly(P<0.01);goblet cells proliferated significantly,mucus secretion increased significantly(P<0.01);the protein expressions of EGFR,PKC,NF-κB p65,MUC5AC and mRNA expressions of EGFR and MUC5AC in lung tissue increased significantly(P<0.01),and the content of MUC5AC in lung tissue increased significantly(P<0.01).Compared with the model group,PEF,TV,EV and EF50 in dexamethasone group and Yifei Jianpi Prescription each dosage groups increased in varying degrees;the pathological injury of lung tissue was alleviated to varying degrees,the score of lung tissue injury was reduced;the proliferation of goblet cells was reduced,and the secretion of mucus was reduced,the expressions of EGFR,PKC,NF-κB p65,MUC5AC protein and EGFR,MUC5AC mRNA in lung tissue decreased,and the content of MUC5AC in lung tissue decreased.There was statistical significance in dexamethasone group and Yifei Jianpi Prescription high-and medium-dosage groups(P<0.01).Conclusion Yifei Jianpi Prescription can inhibit the hypersecretion of airway mucus and the high expression of EGFR/PKC/NF-κB pathway protein in rats with ALI induced by LPS.

Objective To observe the synergistic effect of Shengxian Huaxian Prescription and its disassembled prescription on pulmonary fibrosis;To explore whether its mechanism is related to regulating the STAT6/PPAR-y pathway to promote polarization of M2 type macrophages towards M1 type.Methods Ten SD rats were randomly selected from 70 rats as blank group,and the remaining rats were re-established pulmonary fibrosis model by intratracheal infusion of bleomycin.After modeling,the rats were divided into model group,positive group,Shengxian Huaxian Prescription group,Shengxian group,Tongluo group and Bushen group,with 10 rats in each group.Shengxian Huaxian Prescription group,Shengxian group,Tongluo group and Bushen group were given 12.60,7.65,3.60 and 2.25 g/kg of corresponding TCM solution,respectively;the positive group was given 0.12 g/kg of pirfenidone suspension;the blank group and the model group were given equal volume of normal saline,once a day,for consecutive 28 days.The lung function of rats was detected,the contents of IL-6 and TGF-β1 in serum were detected by ELISA,the pathological changes in lung tissue were observed by Masson staining,the expression of CD68,iNOS and CD206,Arg-1 in lung tissue were detected by immunofluorescence,the expression of SOCS1,SOCS3,STAT6,p-STAT6 and PPAR-γ in lung tissue were detected by Western blot.Results Compared with the blank group,the PEF,PIF and EF50 in model group rats significantly decreased,and the contents of serum IL-6 and TGF-β1 significantly increased,Masson staining showed a large amount of collagen fiber deposition,Ashcroft score significantly increased,CD206,Arg-1,STAT6,p-STAT6,PPAR-y protein expression significantly increased(P＜0.01),the expressions of SOCS1 and SOCS3 protein significantly decreased(P＜0.01),while the expression of CD68 and iNOS were not significantly changed(P＞0.05).Compared with the model group,the PEF,PIF and EF50 in all administration groups significantly increased,and the contents of serum IL-6 and TGF-β1 significantly decreased,collagen fiber deposition in lung tissue were decreased to varying degree,Ashcroft score significantly decreased,the expression of CD206,Arg-1,STAT6,p-STAT6 and PPAR-γ protein significantly decreased(P＜0.01),the expressions of CD68,iNOS,SOCS1 and SOCS3 protein significantly increased(P＜0.05).The above indicators showed the most significant changes in Shengxian Huaxian Prescription group,followed by Shengxian group(P＜0.01,P＜0.05).Conclusion Both Shengxian Huaxian Prescription and its disassembled prescription have anti pulmonary fibrosis effects,and their mechanism may related to regulating the STAT6/PPAR-y pathway and promoting polarization of M2 type macrophages towards M1 type.Shengxian Huaxian Prescription group has the best effect,while Shengxian group play an important role in the prescription,and the compatibility between each group has a synergistic effect.

Pulmonary fibrosis is a respiratory system disorder characterized by damage to alveolar epithelial cells,pathological proliferation and transformation of fibroblasts,excessive deposition of extracellular matrix,leading to structural damage and loss of function in lung tissues,with a high mortality rate and limited effective treatment methods.This article was based on the TCM understanding of"lung connecting to large intestine",namely the theory of"lung and the large intestine being interior-exterior related",and set the modern medical understanding of"lung connecting to large intestine",namely the theory of"gut-lung axis"as the key.Combining the TCM pathogenesis of pulmonary fibrosis and the related mechanisms of"gut-lung axis"in pulmonary fibrosis,it preliminarily expounded the connotation of TCM regulating the"gut-lung axis"to treat pulmonary fibrosis,aiming to provide new ideas for clinical treatment of pulmonary fibrosis through the"gut-lung axis".