Objective To investigate the protective effects of Jiawei Danshen Decoction(加味丹参饮,JWDSD)on myocardial ischemia-reperfusion injury(MIRI)via the regulation of serum Hydrogen sulfide(H2S)and cardiac Beclin1,light Ch...Objective To investigate the protective effects of Jiawei Danshen Decoction(加味丹参饮,JWDSD)on myocardial ischemia-reperfusion injury(MIRI)via the regulation of serum Hydrogen sulfide(H2S)and cardiac Beclin1,light Chain 3 A/B(LC3 A/B),p62,and autophagy protein5(ATG5).Methods Seventy specific pathogen free(SPF)Sprague-Dawley(SD)rats were randomly assigned to seven groups(n=10 in each group),including normal control,sham operation,MIRI model(model),ischemic preconditioning,Na HS,JWDSD,and JWDSD+CSE inhibitor(JWDSD+PPG)groups,and orally administered the indicated drugs for 14 d.Two hours after the last administration,the left anterior decreased branch of the coronary artery of each rat in model,Na HS,JWDSD,and JWDSD+PPG groups was ligated for 30 min and subsequently reperfused for 90 min to establish the MIRI model,and the rats in the sham operation group were only exposed to the thorax after surgery without coronary ligation.Blood samples were collected to detect H2S levels using an enzyme-linked immunosorbent assay(ELISA).Heart tissues were harvested for histopathological and immunohistochemical examination and quantitative reverse transcription polymerase chain reaction analysis of Beclin1 and ATG5 m RNA expression and Western blot analysis of Beclin1,LC3 A/B,and p62 protein expression.Results(1)The serum H2S content in model group rats was significantly reduced(P<0.01),JWDSD significantly increased the serum H2S content of model group rats(P<0.01),and the CSE inhibitor(PPG)significantly reduced H2S levels in the JWDSD group rats(P<0.01).(2)Compared with the normal control group,the myocardial tissue necrosis and cell destruction occurred in the MIRI model group,and JWDSD could alleviate the myocardial tissue necrosis of model rats,but the ameliorative effect of JWDSD could be reversed by PPG.(3)Beclin1,LC3 A/B,and p62 expression levels in the heart tissues of the model group were significantly increased(P<0.001),whereas decreased by JWDSD(P<0.05,P<0.01,and P<0.001,respectively),and the inhibitory effects of JWDSD on Beclin1,LC3 A/B,and p62 expression were partially reversed by PPG(P<0.01,P<0.05,and P<0.01,respectively).(4)The expression levels of autophagy-related genes Beclin1 and ATG5 were significantly increased in the model group(P<0.001).JWDSD clearly downregulated the expression levels of Beclin1 and ATG5(P<0.05 and P<0.001,respectively),which were reversed by PPG(P<0.001).Conclusion Our experimental data show that JWDSD can exhibit an anti-MIRI role by increasing endogenous H2S generation,and downregulating the expression of Beclin1,LC3 A/B,p62 and ATG5,which are related to inhibiting autophagy signaling.展开更多
Objective This study aimed to compare the mechanisms and efficacies of five formulas that improve blood circulation and remove blood stasis.Methods(1)A network pharmacology method was used to determine the targets of ...Objective This study aimed to compare the mechanisms and efficacies of five formulas that improve blood circulation and remove blood stasis.Methods(1)A network pharmacology method was used to determine the targets of five formulas that promote circulation and remove blood stasis.Compounds of the five formulas,namely Danshen Yin(丹参饮,DSY),Huoluo Xiaoling Dan(活络效灵丹,HLXLD),Shixiao San(失笑散,SXS),Taohong Siwu Tang(桃红四物汤,THSWT),and Xuefu Zhuyu Tang(血府逐瘀汤,XFZYT),were retrieved from the Traditonal Chinese Medicine System Pharmacology Database(TCMSP),the Shanghai Institute of Organic Chemistry of CAS,and the TCM Integrated Database.Drug target network was constructed by searching the Swiss Target Prediction database and the STITCH database.The target network of stasis was extracted from the PharmGKB database,the Online Mendelian Inheritance in Man(OMIM)database,the Genetic Association Database(GAD),and the Therapeutic Target Database(TTD).Candidate targets were determined using protein-protein interaction(PPI)network extension and topology selection.Thereafter,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was used to determine the differentiation of the mechanism of the five formulas.(2)Animal experiments were conducted to explore the efficacies of the five formulas in treating blood stasis.Seventy New Zealand rabbits were exposed to high-fat feeding+epinephrine injection to construct a blood stasis syndrome model.The rabbits were evenly divided into control,model,DSY,HLXLD,SXS,THSWT,and XFZYT groups.The latter five groups were orally administered the corresponding formulas[DSY:3.92 g/(kg·d),XFZYT:7.10 g/(kg·d),SXS:1.12 g/(kg·d),HLXLD:5.60 g/(kg·d),THSWT:4.48 g/(kg·d)].Serum lipid and blood rheology were analyzed,and pathology slices were observed.Results(1)A total of 269,358,288,370,and 376 candidate targets of DSY,HLXLD,SXS,THSWT,and XFZYT were obtained among which were 232 shared candidate targets.Fluid shear stress and atherosclerosis were the biological processes common to the five formulas.HLXLD,SXS,DSY,and THSWT regulated lipolysis in adipocytes,and XFZYT,HLXLD,SXS,and THSWT regulated the AGE-RAGE signaling pathway and complement and coagulation cascades.HLXLD,SXS,and XFZYT regulated the HIF-1 signaling pathway,DSY regulated the cGMP-PKG signaling pathway,HLXLD reduced platelet activation,SXS regulated the calcium signaling pathway,and XFZYT regulated the PPAR signaling pathway.(2)In the animal experiments,the values of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL),high density lipoprotein cholesterol(HDL),TC/HDL,and TG/HDL in each group decreased,among which the ones seen in XFZYT,HLXLD,and SXS groups were statistically significant(P<0.05).XFZYT presented the best effect,followed by HLXLD and SXS.XFZYT and HLXLD decreased apolipoprotein B100(apoB100)and increased apolipoprotein A1/apoB100(P<0.05).XFZYT decreased all the values of hematocrit(HCT),plasma viscosity,whole blood viscosity(WBV);HLXLD and SXS affected HCT;and DSY and THSWT regulated WBV(P<0.05).All the five formulas decreased the values of optical density and area of plaque,among which XFZYT and HLXLD showed statistical significance(P<0.05).Conclusion Adjusting fluid shear stress and alleviating the injury of endothelial cells might be the common mechanisms by which the five formulas promote blood circulation and remove blood stasis.Different formulas also have unique targets,which may provide guidance for clinical drug selection.By regulating different indices,the five formulas can regulate blood lipid and hemorheology,improve the state of blood stasis,and decrease the degree of aortic plaque in the blood stasis model rabbits.XFZYT and HLXLD had higher efficacies than DSY,THSWT,and SXS.展开更多
基金funding support from the National Natural Science Foundation of China(No.81704065)Hunan Provincial Natural Science Foundation(No.2019JJ40225)+1 种基金the Scientific Research Project of Education Department of Hunan Province(No.19B415,No.19C1393 and No.20C1392)Hunan Provincial Scientific Research Project of Chinese Medicine(No.2020015)。
文摘Objective To investigate the protective effects of Jiawei Danshen Decoction(加味丹参饮,JWDSD)on myocardial ischemia-reperfusion injury(MIRI)via the regulation of serum Hydrogen sulfide(H2S)and cardiac Beclin1,light Chain 3 A/B(LC3 A/B),p62,and autophagy protein5(ATG5).Methods Seventy specific pathogen free(SPF)Sprague-Dawley(SD)rats were randomly assigned to seven groups(n=10 in each group),including normal control,sham operation,MIRI model(model),ischemic preconditioning,Na HS,JWDSD,and JWDSD+CSE inhibitor(JWDSD+PPG)groups,and orally administered the indicated drugs for 14 d.Two hours after the last administration,the left anterior decreased branch of the coronary artery of each rat in model,Na HS,JWDSD,and JWDSD+PPG groups was ligated for 30 min and subsequently reperfused for 90 min to establish the MIRI model,and the rats in the sham operation group were only exposed to the thorax after surgery without coronary ligation.Blood samples were collected to detect H2S levels using an enzyme-linked immunosorbent assay(ELISA).Heart tissues were harvested for histopathological and immunohistochemical examination and quantitative reverse transcription polymerase chain reaction analysis of Beclin1 and ATG5 m RNA expression and Western blot analysis of Beclin1,LC3 A/B,and p62 protein expression.Results(1)The serum H2S content in model group rats was significantly reduced(P<0.01),JWDSD significantly increased the serum H2S content of model group rats(P<0.01),and the CSE inhibitor(PPG)significantly reduced H2S levels in the JWDSD group rats(P<0.01).(2)Compared with the normal control group,the myocardial tissue necrosis and cell destruction occurred in the MIRI model group,and JWDSD could alleviate the myocardial tissue necrosis of model rats,but the ameliorative effect of JWDSD could be reversed by PPG.(3)Beclin1,LC3 A/B,and p62 expression levels in the heart tissues of the model group were significantly increased(P<0.001),whereas decreased by JWDSD(P<0.05,P<0.01,and P<0.001,respectively),and the inhibitory effects of JWDSD on Beclin1,LC3 A/B,and p62 expression were partially reversed by PPG(P<0.01,P<0.05,and P<0.01,respectively).(4)The expression levels of autophagy-related genes Beclin1 and ATG5 were significantly increased in the model group(P<0.001).JWDSD clearly downregulated the expression levels of Beclin1 and ATG5(P<0.05 and P<0.001,respectively),which were reversed by PPG(P<0.001).Conclusion Our experimental data show that JWDSD can exhibit an anti-MIRI role by increasing endogenous H2S generation,and downregulating the expression of Beclin1,LC3 A/B,p62 and ATG5,which are related to inhibiting autophagy signaling.
基金We thank for the funding support from the Special Project of Central Government Guiding Local Science and Technology Development(No.2019XF5062)the 211 Project of Chinese Medicine Diagnostics(No.80019992)the Open Fund of National Key Discipline of Chinese Medicine Diagnostics of Hunan University of Chinese Medicine(No.ZZKF201501 and No.2015ZYZD01).
文摘Objective This study aimed to compare the mechanisms and efficacies of five formulas that improve blood circulation and remove blood stasis.Methods(1)A network pharmacology method was used to determine the targets of five formulas that promote circulation and remove blood stasis.Compounds of the five formulas,namely Danshen Yin(丹参饮,DSY),Huoluo Xiaoling Dan(活络效灵丹,HLXLD),Shixiao San(失笑散,SXS),Taohong Siwu Tang(桃红四物汤,THSWT),and Xuefu Zhuyu Tang(血府逐瘀汤,XFZYT),were retrieved from the Traditonal Chinese Medicine System Pharmacology Database(TCMSP),the Shanghai Institute of Organic Chemistry of CAS,and the TCM Integrated Database.Drug target network was constructed by searching the Swiss Target Prediction database and the STITCH database.The target network of stasis was extracted from the PharmGKB database,the Online Mendelian Inheritance in Man(OMIM)database,the Genetic Association Database(GAD),and the Therapeutic Target Database(TTD).Candidate targets were determined using protein-protein interaction(PPI)network extension and topology selection.Thereafter,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was used to determine the differentiation of the mechanism of the five formulas.(2)Animal experiments were conducted to explore the efficacies of the five formulas in treating blood stasis.Seventy New Zealand rabbits were exposed to high-fat feeding+epinephrine injection to construct a blood stasis syndrome model.The rabbits were evenly divided into control,model,DSY,HLXLD,SXS,THSWT,and XFZYT groups.The latter five groups were orally administered the corresponding formulas[DSY:3.92 g/(kg·d),XFZYT:7.10 g/(kg·d),SXS:1.12 g/(kg·d),HLXLD:5.60 g/(kg·d),THSWT:4.48 g/(kg·d)].Serum lipid and blood rheology were analyzed,and pathology slices were observed.Results(1)A total of 269,358,288,370,and 376 candidate targets of DSY,HLXLD,SXS,THSWT,and XFZYT were obtained among which were 232 shared candidate targets.Fluid shear stress and atherosclerosis were the biological processes common to the five formulas.HLXLD,SXS,DSY,and THSWT regulated lipolysis in adipocytes,and XFZYT,HLXLD,SXS,and THSWT regulated the AGE-RAGE signaling pathway and complement and coagulation cascades.HLXLD,SXS,and XFZYT regulated the HIF-1 signaling pathway,DSY regulated the cGMP-PKG signaling pathway,HLXLD reduced platelet activation,SXS regulated the calcium signaling pathway,and XFZYT regulated the PPAR signaling pathway.(2)In the animal experiments,the values of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL),high density lipoprotein cholesterol(HDL),TC/HDL,and TG/HDL in each group decreased,among which the ones seen in XFZYT,HLXLD,and SXS groups were statistically significant(P<0.05).XFZYT presented the best effect,followed by HLXLD and SXS.XFZYT and HLXLD decreased apolipoprotein B100(apoB100)and increased apolipoprotein A1/apoB100(P<0.05).XFZYT decreased all the values of hematocrit(HCT),plasma viscosity,whole blood viscosity(WBV);HLXLD and SXS affected HCT;and DSY and THSWT regulated WBV(P<0.05).All the five formulas decreased the values of optical density and area of plaque,among which XFZYT and HLXLD showed statistical significance(P<0.05).Conclusion Adjusting fluid shear stress and alleviating the injury of endothelial cells might be the common mechanisms by which the five formulas promote blood circulation and remove blood stasis.Different formulas also have unique targets,which may provide guidance for clinical drug selection.By regulating different indices,the five formulas can regulate blood lipid and hemorheology,improve the state of blood stasis,and decrease the degree of aortic plaque in the blood stasis model rabbits.XFZYT and HLXLD had higher efficacies than DSY,THSWT,and SXS.
