ObjectiveTo investigate the anti-inflammatory effect and mechanism of Danggui Shaoyaosan （DSS） in ameliorating vascular dementia （VaD）. MethodsSeventy 8-week-old C57BL/6J male mice were randomized into 7 groups： control， model， sham， positive control （donepezil， 10 mg·kg^-1）， and low-， medium-， and high-dose （12， 24， 36 g·kg^-1， respectively） DSS groups （n=10）. After drug administration， behavioral tests and cerebral blood flow detection were carried out. Enzyme-linked immunosorbent assay was used to assess the levels of interferon （IFN）-α， tumor necrosis factor （TNF）-α， interleukin （IL）-6， and CXC motif chemokine ligand 10 （CXCL10） in the brain tissue. Hematoxylin-eosin staining was employed to observe the changes in hippocampal morphology. Transcriptomics was used to predict the potential signaling mechanisms of DSS in ameliorating neuroinflammation， and Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were conducted to verify the expression changes of related genes. ResultsCompared with the control group and the sham group， the model group showed deceases in recognition index in the new object recognition test， movement distance and time in the target quadrant， and number of crossings in the Morris water maze test （P<0.01）. In addition， the model group showed slow cerebral blood flow and down-regulated protein levels of postsynaptic density protein 95 （PSD95） and occludin （P<0.05， P<0.01）， and there was no significant difference between the control group and the sham group. Compared with the model group， the DSS at each dose increased the new object recognition index， the distance and time in the target quadrant， the number of crossings， and cerebral blood flow （P<0.05， P<0.01）. In terms of brain tissue injury-related protein expression and inflammatory factor content， the medium-dose DSS group had the best effect， with higher protein levels of PSD95 and occludin （P<0.05） and lower levels of IFN-α， TNF-α， and IL-6 （P<0.01） than the model group. The hippocampus of model mice showed pathological manifestations such as cell loss and disarrangement， which were alleviated after administration of DSS at each dose. Transcriptomic results indicated that interferon regulatory factor 7 （IRF7）， signal transducer and activator of transcription 3 （STAT3）， and bone marrow stromal cell antigen 2 （BST2） were differentially expressed genes （down-regulated） in control group and medium-dose DSS group compared with the model group. The KEGG pathway enrichment analysis showed that RIG-Ⅰ-like receptor signaling pathway， Toll-like receptor signaling pathway， cytosolic DNA-sensing pathway， and downstream stimulator of interferon genes （STING） were both upstream signaling pathways affecting IFN expression. Real-time PCR results indicated that the mRNA levels of cyclic good manufacturing practice（GMP）-adenosine monophosphate（AMP） synthase （cGAS）， STING， IRF7， STAT3， and BST2 in the model group were higher than those in the sham group （P<0.05， P<0.01）. The mRNA levels of all the genes in the medium-dose DSS group were down-regulated compared with those in the model group after administration （P<0.05， P<0.01）. Western blot results indicated that the relative expression levels of cGAS， STING， p-IRF7， p-STAT3， and BST2 in the model group were higher than those in the sham group （P<0.05， P<0.01）. The protein levels of cGAS， STING， p-IRF7， p-STAT3， and BST2 in the medium-dose DSS group were decreased compared with those in the model group （P<0.05， P<0.01）. ConclusionDSS ameliorates cognitive impairment in the VaD model mice by inhibiting the cGAS-STING/IRF7/STAT3-mediated neuroinflammation.

ObjectiveTo investigate the anti-inflammatory effect and mechanism of Danggui Shaoyaosan （DSS） in ameliorating vascular dementia （VaD）. MethodsSeventy 8-week-old C57BL/6J male mice were randomized into 7 groups： control， model， sham， positive control （donepezil， 10 mg·kg^-1）， and low-， medium-， and high-dose （12， 24， 36 g·kg^-1， respectively） DSS groups （n=10）. After drug administration， behavioral tests and cerebral blood flow detection were carried out. Enzyme-linked immunosorbent assay was used to assess the levels of interferon （IFN）-α， tumor necrosis factor （TNF）-α， interleukin （IL）-6， and CXC motif chemokine ligand 10 （CXCL10） in the brain tissue. Hematoxylin-eosin staining was employed to observe the changes in hippocampal morphology. Transcriptomics was used to predict the potential signaling mechanisms of DSS in ameliorating neuroinflammation， and Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were conducted to verify the expression changes of related genes. ResultsCompared with the control group and the sham group， the model group showed deceases in recognition index in the new object recognition test， movement distance and time in the target quadrant， and number of crossings in the Morris water maze test （P<0.01）. In addition， the model group showed slow cerebral blood flow and down-regulated protein levels of postsynaptic density protein 95 （PSD95） and occludin （P<0.05， P<0.01）， and there was no significant difference between the control group and the sham group. Compared with the model group， the DSS at each dose increased the new object recognition index， the distance and time in the target quadrant， the number of crossings， and cerebral blood flow （P<0.05， P<0.01）. In terms of brain tissue injury-related protein expression and inflammatory factor content， the medium-dose DSS group had the best effect， with higher protein levels of PSD95 and occludin （P<0.05） and lower levels of IFN-α， TNF-α， and IL-6 （P<0.01） than the model group. The hippocampus of model mice showed pathological manifestations such as cell loss and disarrangement， which were alleviated after administration of DSS at each dose. Transcriptomic results indicated that interferon regulatory factor 7 （IRF7）， signal transducer and activator of transcription 3 （STAT3）， and bone marrow stromal cell antigen 2 （BST2） were differentially expressed genes （down-regulated） in control group and medium-dose DSS group compared with the model group. The KEGG pathway enrichment analysis showed that RIG-Ⅰ-like receptor signaling pathway， Toll-like receptor signaling pathway， cytosolic DNA-sensing pathway， and downstream stimulator of interferon genes （STING） were both upstream signaling pathways affecting IFN expression. Real-time PCR results indicated that the mRNA levels of cyclic good manufacturing practice（GMP）-adenosine monophosphate（AMP） synthase （cGAS）， STING， IRF7， STAT3， and BST2 in the model group were higher than those in the sham group （P<0.05， P<0.01）. The mRNA levels of all the genes in the medium-dose DSS group were down-regulated compared with those in the model group after administration （P<0.05， P<0.01）. Western blot results indicated that the relative expression levels of cGAS， STING， p-IRF7， p-STAT3， and BST2 in the model group were higher than those in the sham group （P<0.05， P<0.01）. The protein levels of cGAS， STING， p-IRF7， p-STAT3， and BST2 in the medium-dose DSS group were decreased compared with those in the model group （P<0.05， P<0.01）. ConclusionDSS ameliorates cognitive impairment in the VaD model mice by inhibiting the cGAS-STING/IRF7/STAT3-mediated neuroinflammation.

Objective To explore the mechanism of Huoxin pill(HXP)in the prevention and treatment of heart failure(HF)based on transcriptomics and network pharmacology.Methods The mice were randomly divided into the normal control group,model control group,positive control group treated with sacubitril/valsartan(60 mg·kg-1),low-dose group treated with HXP(31.2 mg·kg-1),and high-dose group treated with HXP(62.4 mg·kg-1).The model control group and each drug treat-ment group were subcutaneously injected with an equal volume of ISO(5 mg·kg-1)for modeling,while the normal control group was given an equal volume of sterile saline.Six hours later,each drug administration group was gavaged with the corresponding drug for intervention,and the normal control and model control groups were gavaged with an equal volume of sterile water.The modeling and drug administration were continued for 21 days.The cardiac function parameters of the mice were measured using color Doppler ultrasound imaging;ELISA was used to detect the levels of mouse serum cAMP,NT-proBNP,and BNP;HE staining and Masson's trichrome staining were used to evaluate the pathological morphology of cardiac tissue,and the CVF was calculated.Network pharmacology combined with transcriptomics was used to predict potential targets and signaling pathways of HXP in the prevention and treatment of HF,and molecular biology methods were used for validation.Results Compared with the normal control group,the model control group showed an increase in LVESd and LVEDd(P＜0.01),and a decrease in LVEF and LVFS(P＜0.01);BNP,NT-proBNP,and cAMP levels were increased(P＜0.01);myocardial collagen fibers increased and CVF in-creased(P＜0.01).Compared with the model control group,the HXP low-dose group,HXP high-dose group,and positive control groups showed a decrease in LVESd and LVEDd(P＜0.01),and an increase in LVEF and LVFS(P＜0.01);serum levels of BNP,NT-proBNP,and cAMP decreased(P＜0.05);the degree of myocardial fibrosis decreased and CVF decreased(P＜0.01).Network pharmacology combined with transcriptomics predicted 10 key targets for HXP in the prevention and treatment of HF:CACNA1H,SCN10A,FGF12,PVALB,ACAN,LGALS3,SERPINE1,MMP3,GSTM1,VDR.Western blot results showed that the protein activation levels of PKA and CREB in myocardial tissue were increased in the model control group compared with the nor-mal control group(P＜0.01).Compared with the model control group,HXP low-dose group、HXP high-dose group,and positive control groups showed a decrease in the protein activation levels of PKA and CREB in myocardial tissue(P＜0.05).Conclusion HXP has an improvement effect on ISO-induced HF in mice,which may involve numerous targets and the cAMP/PKA signaling pathway.

Objective To explore the mechanism of Huoxin pill(HXP)in the prevention and treatment of heart failure(HF)based on transcriptomics and network pharmacology.Methods The mice were randomly divided into the normal control group,model control group,positive control group treated with sacubitril/valsartan(60 mg·kg-1),low-dose group treated with HXP(31.2 mg·kg-1),and high-dose group treated with HXP(62.4 mg·kg-1).The model control group and each drug treat-ment group were subcutaneously injected with an equal volume of ISO(5 mg·kg-1)for modeling,while the normal control group was given an equal volume of sterile saline.Six hours later,each drug administration group was gavaged with the corresponding drug for intervention,and the normal control and model control groups were gavaged with an equal volume of sterile water.The modeling and drug administration were continued for 21 days.The cardiac function parameters of the mice were measured using color Doppler ultrasound imaging;ELISA was used to detect the levels of mouse serum cAMP,NT-proBNP,and BNP;HE staining and Masson's trichrome staining were used to evaluate the pathological morphology of cardiac tissue,and the CVF was calculated.Network pharmacology combined with transcriptomics was used to predict potential targets and signaling pathways of HXP in the prevention and treatment of HF,and molecular biology methods were used for validation.Results Compared with the normal control group,the model control group showed an increase in LVESd and LVEDd(P＜0.01),and a decrease in LVEF and LVFS(P＜0.01);BNP,NT-proBNP,and cAMP levels were increased(P＜0.01);myocardial collagen fibers increased and CVF in-creased(P＜0.01).Compared with the model control group,the HXP low-dose group,HXP high-dose group,and positive control groups showed a decrease in LVESd and LVEDd(P＜0.01),and an increase in LVEF and LVFS(P＜0.01);serum levels of BNP,NT-proBNP,and cAMP decreased(P＜0.05);the degree of myocardial fibrosis decreased and CVF decreased(P＜0.01).Network pharmacology combined with transcriptomics predicted 10 key targets for HXP in the prevention and treatment of HF:CACNA1H,SCN10A,FGF12,PVALB,ACAN,LGALS3,SERPINE1,MMP3,GSTM1,VDR.Western blot results showed that the protein activation levels of PKA and CREB in myocardial tissue were increased in the model control group compared with the nor-mal control group(P＜0.01).Compared with the model control group,HXP low-dose group、HXP high-dose group,and positive control groups showed a decrease in the protein activation levels of PKA and CREB in myocardial tissue(P＜0.05).Conclusion HXP has an improvement effect on ISO-induced HF in mice,which may involve numerous targets and the cAMP/PKA signaling pathway.