Objective:To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula(HTQS for-mula),for the health management and treatment of non-alcoholic fatty liver disease(NAFLD).Methods:A rat model of NAFLD was ...Objective:To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula(HTQS for-mula),for the health management and treatment of non-alcoholic fatty liver disease(NAFLD).Methods:A rat model of NAFLD was employed to examine the efficacy and safety of the HTQS formula.In vivo active components and potential mechanisms of the HTQS formula were identified using UPLC‒MS/MS combined with network pharmacology.The influence of the HTQS formula on the dominating proteins in PI3K/Akt pathway was validated in vivo using western blot.Finally,16S rRNA sequencing of the gut microbiome was conducted followed by targeted metabolomics detecting fecal short-chain fatty acids(SCFAs)and bile acids to determine the impact of the HTQS formula on gut microbiota.Results:The HTQS formula reduced weight gain and hepatic steatosis in NAFLD rats and decreased serum total cholesterol(TC),triglycerides,blood glucose,and insulin resistance(IR)without causing liver or kidney injury.We detected 28 components using UPLC‒MS/MS and identified 439 shared targets be-tween NAFLD and the HTQS formula.Primarily,we focused on the PI3K/Akt signaling pathway based on protein‒protein interaction network analysis.We validated that the HTQS formula inhibited liver stea-tosis and inflammation by increasing the phosphorylation levels of PI3K,AKT,P27,GSK3b in the PI3K/Akt signaling pathway.16S rRNA sequencing revealed that the HTQS formula reduced the abundance of the genus Family_XIII_AD3011_group,which was positively correlated with IR and taurodeoxycholic acid.In addition,Lachnospiraceae_UCG_010 inversely correlated with TC and five bile acids,which could be essential to the therapeutic effect of the HTQS formula against NAFLD.Conclusions:The HTQS formula proved to be an effective pharmacotherapy for NAFLD without causing liver or kidney injury.Multiple potent components of the HTQS formula could alleviate liver steatosis and lipid metabolism disorder by modulating the PI3K/Akt signaling pathway and restoring gut microbiota composition.展开更多
Objective This study aimed to analyze the mechanism of action of the Pingyang Jiangya Formula(平阳降压方,PYJYF)in treating hypertension,based on network pharmacology,and to verify the subsequent predictions through an...Objective This study aimed to analyze the mechanism of action of the Pingyang Jiangya Formula(平阳降压方,PYJYF)in treating hypertension,based on network pharmacology,and to verify the subsequent predictions through animal experiments.Methods The active components and related target genes of PYJYF were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM),Encyclopedia of Traditional Chinese Medicine(ETCM),and Drug Bank databases and available literature.The hypertension target genes were screened based on Therapeutic Target Database(TTD),GeneCards,Online Mendelian Inheritance in Man(OMIM),UniProt,and relevant literature.The component-disease-target network intersection target genes were inputted into the STRING database,and the key target genes were selected according to the degree algorithm.Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed to explore the multitarget mechanism of action and molecular regulatory network of PYJYF in the treatment of hypertension.To verify this prediction,we used PYJYF to intervene in spontaneously hypertensive rats(SHRs)and Wistar–Kyoto rats(WKY)as normal control,and the noninvasive tail artery manometry method was used to measure systolic blood pressure(SBP)in the rat tail before PYJYF intervention.After drug intervention,the SBP of each group rats were measured and compared every week.Enzyme-linked immunosorbent assay(ELISA)was used to test plasma renin,angiotensin II(Ang II),and aldosterone(Ald)levels,and hematoxylin-eosin(HE)staining was used to observe pathological damage to the renal vessels in each group of rats.Western blot and reverse transcription real-time quantitative PCR(RT-PCR)were used to detect the protein and mRNA expression levels of PI3 K,AKT1,BAX,and Bcl-2,respectively.Results A total of 4123 hypertension targets were obtained from related databases.From the TCMSP and chemical databases,78 active components of PYJYF and the corresponding 401 drug targets were retrieved.Data analysis revealed that 208 drug targets directly interacted with the hypertension targets in PYJYF.The 10 targets most closely related to hypertension target proteins in PYJYF were directly retrieved from relevant databases.GO analysis revealed that 10 direct target proteins were involved in all aspects of the antihypertensive effects of PYJYF,as well as molecular biological processes,such as the regulation of blood pressure,renin-angiotensin-aldosterone system(RAAS),angiotensin-mediated ligand reactions,and biological stimulation of cardiomyocyte apoptosis.KEGG pathway enrichment analysis revealed that PYJYF directly affected 20 signaling pathways associated with hypertension.In animal experiments,PYJYF reduced the protein and m RNA levels of PI3 K,Akt,and Bax and upregulated the expression of the protein and m RNA levels of Bcl-2,reduced plasma renin,Ang II,and Ald levels,improved the hyperactivity of RAAS,and significantly reduced SBP in SHRs.Conclusion PYJYF is effective for hypertension therapy that acts through multiple compounds and targets.The possible underlying molecular mechanism includes regulating the PI3 K/Akt signaling pathway to suppress RAAS,increasing the ratio of Bcl-2/Bax proteins,and inhibiting apoptosis,thereby mediating the repair of renal and renal vascular damage caused by hypertension.These findings warrant further research for use in clinical settings.展开更多
基金supported by the General Program of National Natural Science Foundation of China(82374308)National Key Research and Development Program(NKRDP)(2022YFC2010104)Henan Province Special Projects of Traditional Chinese Medicine Science Research(2024ZY2067),and National Talent Precision Cultivation Plan of the Beijing University of Chinese Medicine.
文摘Objective:To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula(HTQS for-mula),for the health management and treatment of non-alcoholic fatty liver disease(NAFLD).Methods:A rat model of NAFLD was employed to examine the efficacy and safety of the HTQS formula.In vivo active components and potential mechanisms of the HTQS formula were identified using UPLC‒MS/MS combined with network pharmacology.The influence of the HTQS formula on the dominating proteins in PI3K/Akt pathway was validated in vivo using western blot.Finally,16S rRNA sequencing of the gut microbiome was conducted followed by targeted metabolomics detecting fecal short-chain fatty acids(SCFAs)and bile acids to determine the impact of the HTQS formula on gut microbiota.Results:The HTQS formula reduced weight gain and hepatic steatosis in NAFLD rats and decreased serum total cholesterol(TC),triglycerides,blood glucose,and insulin resistance(IR)without causing liver or kidney injury.We detected 28 components using UPLC‒MS/MS and identified 439 shared targets be-tween NAFLD and the HTQS formula.Primarily,we focused on the PI3K/Akt signaling pathway based on protein‒protein interaction network analysis.We validated that the HTQS formula inhibited liver stea-tosis and inflammation by increasing the phosphorylation levels of PI3K,AKT,P27,GSK3b in the PI3K/Akt signaling pathway.16S rRNA sequencing revealed that the HTQS formula reduced the abundance of the genus Family_XIII_AD3011_group,which was positively correlated with IR and taurodeoxycholic acid.In addition,Lachnospiraceae_UCG_010 inversely correlated with TC and five bile acids,which could be essential to the therapeutic effect of the HTQS formula against NAFLD.Conclusions:The HTQS formula proved to be an effective pharmacotherapy for NAFLD without causing liver or kidney injury.Multiple potent components of the HTQS formula could alleviate liver steatosis and lipid metabolism disorder by modulating the PI3K/Akt signaling pathway and restoring gut microbiota composition.
基金funding support from the National Natural Science Foundation of China(No.81874464)National Major New Drug Development Project(No.2019ZX09301-103)Provincial Department of Graduate Research Innovation Project of Hunan(No.CX20190565)。
文摘Objective This study aimed to analyze the mechanism of action of the Pingyang Jiangya Formula(平阳降压方,PYJYF)in treating hypertension,based on network pharmacology,and to verify the subsequent predictions through animal experiments.Methods The active components and related target genes of PYJYF were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM),Encyclopedia of Traditional Chinese Medicine(ETCM),and Drug Bank databases and available literature.The hypertension target genes were screened based on Therapeutic Target Database(TTD),GeneCards,Online Mendelian Inheritance in Man(OMIM),UniProt,and relevant literature.The component-disease-target network intersection target genes were inputted into the STRING database,and the key target genes were selected according to the degree algorithm.Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed to explore the multitarget mechanism of action and molecular regulatory network of PYJYF in the treatment of hypertension.To verify this prediction,we used PYJYF to intervene in spontaneously hypertensive rats(SHRs)and Wistar–Kyoto rats(WKY)as normal control,and the noninvasive tail artery manometry method was used to measure systolic blood pressure(SBP)in the rat tail before PYJYF intervention.After drug intervention,the SBP of each group rats were measured and compared every week.Enzyme-linked immunosorbent assay(ELISA)was used to test plasma renin,angiotensin II(Ang II),and aldosterone(Ald)levels,and hematoxylin-eosin(HE)staining was used to observe pathological damage to the renal vessels in each group of rats.Western blot and reverse transcription real-time quantitative PCR(RT-PCR)were used to detect the protein and mRNA expression levels of PI3 K,AKT1,BAX,and Bcl-2,respectively.Results A total of 4123 hypertension targets were obtained from related databases.From the TCMSP and chemical databases,78 active components of PYJYF and the corresponding 401 drug targets were retrieved.Data analysis revealed that 208 drug targets directly interacted with the hypertension targets in PYJYF.The 10 targets most closely related to hypertension target proteins in PYJYF were directly retrieved from relevant databases.GO analysis revealed that 10 direct target proteins were involved in all aspects of the antihypertensive effects of PYJYF,as well as molecular biological processes,such as the regulation of blood pressure,renin-angiotensin-aldosterone system(RAAS),angiotensin-mediated ligand reactions,and biological stimulation of cardiomyocyte apoptosis.KEGG pathway enrichment analysis revealed that PYJYF directly affected 20 signaling pathways associated with hypertension.In animal experiments,PYJYF reduced the protein and m RNA levels of PI3 K,Akt,and Bax and upregulated the expression of the protein and m RNA levels of Bcl-2,reduced plasma renin,Ang II,and Ald levels,improved the hyperactivity of RAAS,and significantly reduced SBP in SHRs.Conclusion PYJYF is effective for hypertension therapy that acts through multiple compounds and targets.The possible underlying molecular mechanism includes regulating the PI3 K/Akt signaling pathway to suppress RAAS,increasing the ratio of Bcl-2/Bax proteins,and inhibiting apoptosis,thereby mediating the repair of renal and renal vascular damage caused by hypertension.These findings warrant further research for use in clinical settings.
