目的:观察新活络效灵丹对ApoE基因敲除小鼠AS早期斑块的病理形态学改变及转化生长因子-β(TGF-β)表达的影响,探讨新活络效灵丹抗AS早期斑块的可能机制。方法:采用高脂饲料喂养ApoE基因敲除小鼠建立AS模型,将45只小鼠随机分为模型组、...目的:观察新活络效灵丹对ApoE基因敲除小鼠AS早期斑块的病理形态学改变及转化生长因子-β(TGF-β)表达的影响,探讨新活络效灵丹抗AS早期斑块的可能机制。方法:采用高脂饲料喂养ApoE基因敲除小鼠建立AS模型,将45只小鼠随机分为模型组、西药组、中药组,另将C57BL/6J小鼠10只设为空白对照组。在造模的同时即给予药物干预,空白对照组不予处理,模型组予以生理盐水0.3 m L/d,中药组给予新活络效灵丹9.67 g/(kg·d),西药组给予阿托伐他汀钙片3.33 mg/(kg·d),均每日灌胃1次,连续给药13周。取主动脉根部,HE染色进行病理观察,采用RT-PCR技术检测各组TGF-βmRNA的表达水平。结果:模型组主动脉内形成明显的AS斑块,并出现脂质核心,泡沫细胞增多,大量的巨噬细胞的浸润;西药组管壁可见AS斑块,斑块面积明显小于模型组;中药组斑块面积显著减少,部分内膜较为光滑。模型组小鼠主动脉组织中TGF-βmRNA表达水平明显低于空白对照组(P<0.01);西药组、中药组小鼠主动脉组织中TGF-βmRNA表达水平明显高于模型对照组(P<0.05或P<0.01),而中药组小鼠主动脉组织中TGF-βmRNA表达水平高于西药组,但2组比较差异无统计学意义(P>0.05)。结论:新活络效灵丹在一定程度上可通过上调保护性因子TGF-β以抑制免疫炎性反应,发挥抗动脉粥样硬化的作用,可能是其防治AS早期斑块的形成机制之一。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
文摘目的:观察新活络效灵丹对ApoE基因敲除小鼠AS早期斑块的病理形态学改变及转化生长因子-β(TGF-β)表达的影响,探讨新活络效灵丹抗AS早期斑块的可能机制。方法:采用高脂饲料喂养ApoE基因敲除小鼠建立AS模型,将45只小鼠随机分为模型组、西药组、中药组,另将C57BL/6J小鼠10只设为空白对照组。在造模的同时即给予药物干预,空白对照组不予处理,模型组予以生理盐水0.3 m L/d,中药组给予新活络效灵丹9.67 g/(kg·d),西药组给予阿托伐他汀钙片3.33 mg/(kg·d),均每日灌胃1次,连续给药13周。取主动脉根部,HE染色进行病理观察,采用RT-PCR技术检测各组TGF-βmRNA的表达水平。结果:模型组主动脉内形成明显的AS斑块,并出现脂质核心,泡沫细胞增多,大量的巨噬细胞的浸润;西药组管壁可见AS斑块,斑块面积明显小于模型组;中药组斑块面积显著减少,部分内膜较为光滑。模型组小鼠主动脉组织中TGF-βmRNA表达水平明显低于空白对照组(P<0.01);西药组、中药组小鼠主动脉组织中TGF-βmRNA表达水平明显高于模型对照组(P<0.05或P<0.01),而中药组小鼠主动脉组织中TGF-βmRNA表达水平高于西药组,但2组比较差异无统计学意义(P>0.05)。结论:新活络效灵丹在一定程度上可通过上调保护性因子TGF-β以抑制免疫炎性反应,发挥抗动脉粥样硬化的作用,可能是其防治AS早期斑块的形成机制之一。
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.
