| Objective: To clarify the clinical efficacy of Mingjun Powder in the treatment of coronary heart disease with stable angina pectoris(yin cold stagnation syndrome),through network pharmacology prediction and animal experiment verification,to preliminarily explore the mechanism of Mingjun Powder in anti atherosclerosis,so as to provide an objective experimental basis for the application of Mingjun Powder in the clinical treatment of coronary heart disease.Methods:1.Clinical study: 72 patients were collected from January 2022 to December 2022 who attended the outpatient clinic and treatment area of the Department of Cardiology of Changchun Hospital of Traditional Chinese Medicine,with the case type of stable angina pectoris and the Chinese medical evidence of yin-cold stagnation,and they were randomly divided into 36 cases each in the test group and the control group,of which 2 cases were shed in the control group,2 cases were shed in the test group and 1 case was excluded,and finally the total number of patients actually included in the statistical analysis was 67 cases were actually included in the statistical analysis.Patients in both groups were treated with basic Western medicine according to the guideline standards,and the test group was given Mingjun San in combination,and the course of treatment was 4 weeks in both groups.The changes of relevant indexes before and after treatment were evaluated,including angina pectoris efficacy,Seattle Angina Assessment Scale,ECG efficacy,nitroglycerin discontinuation rate,TCM evidence score,blood lipid level and inflammation index hs-CRP level.2.Network pharmacology: screen the active ingredients and action targets of Mingjun Powder through TCMSP database,obtain atherosclerosis targets using Gene Cards,OMIM and Dis GENEN databases,screen intersection targets of Mingjun Powder and atherosclerosis using Uniprot database,construct PPI network using STRING database and Cytoscape software,and conduct enrichment analysis of GO function and KEGG pathway with DAVID database.3.Experimental study: 70 SPF male SD rats were randomly selected as the blank group and given conventional chow,while the remaining 55 rats were given high-fat chow combined with vitamin D3 as the modeling group to establish the AS model,and the modeling period was 9 weeks.The typical pathological changes of AS were observed to ensure the successful establishment of the model.The rats with successful modeling were randomly divided into 5 groups: model group,western medicine group,Mingjun san low-dose group,Mingjun san medium-dose group and Mingjun san high-dose group,10 rats in each group.The blank group and model group were given equal volume of saline gavage,the western medicine group was given atorvastatin calcium tablets solution gavage,and the Mingjun san low,medium and high-dose groups were given different concentrations of Mingjun san solution gavage respectively.The serum levels of TC,TG,HDL-C and LDL-C were measured.The morphological changes of rat thoracic aorta were observed by HE staining;serum NO level was detected by nitrate reduction method;serum TNF-α,IL-6,IL-1β and ET-1 expression levels were detected by ELISA;RT-PCR detected m RNA expression levels of aortic TLR4,MyD88,and NF-κB;aortic TLR4,MyD88 and NF-κB protein expression levels were detected by Western Blot.Results:1.Clinical study:(1)Effect of angina pectoris: after treatment,the total effective rate of angina pectoris in the test group(87.88%)was higher than that in the control group(76.47%),and the difference between the groups was statistically significant(P<0.05);After treatment,the score of angina pectoris symptoms in both groups decreased(P<0.05 or 0.01),and the test group was better than the control group(P<0.05 or 0.01).(2)SAQ score: After treatment,SAQ scores of patients in both groups increased(P<0.05 or 0.01),and the improvement of SAQ scores in the test group was better than that in the control group(P<0.05 or 0.01).(3)ECG efficacy: After treatment,the total effective rate of ECG in the test group(51.52%)was higher than that in the control group(32.35%),but there was no statistically significant difference between the two groups(P>0.05).(4)Nitroglycerin withdrawal rate: after treatment,the Nitroglycerin withdrawal rate in the test group(81.82%)was higher than that in the control group(61.76%),and the difference between the two groups was statistically significant(P<0.05).(5)The curative effect of TCM syndrome: the total effective rate of TCM syndrome in the experimental group(90.91%)was higher than that in the control group(70.59%),the difference was statistically significant(P<0.05);After treatment,the TCM symptom score of both groups decreased(P<0.05 or 0.01);Compared with the control group,the test group was superior to the control group in improving chest pain,chest tightness,shortness of breath,and chilliness(P<0.05 or 0.01);The improvement of the total score of TCM symptoms in the test group was better than that in the control group(P<0.01).(6)Blood lipid level: After treatment,the levels of TG,TC and LDL-C in the test group decreased,while the levels of HDL-C increased(P<0.05 or 0.01);The levels of TC and LDL-C in the control group decreased after treatment(P<0.05);Compared between groups,the levels of TC and LDL-C in the test group were lower than those in the control group,while the levels of HDL-C were higher than those in the control group(P<0.05 or 0.01).(7)After treatment,the level of inflammation index hs-CRP in both groups decreased(P<0.01),and the level of hs-CRP in the test group was lower than that in the control group(P<0.01).2.Network pharmacology: There are 124 effective active ingredients and 221 action targets in Mingjun Powder,and the key ingredients mainly include quercetin β-Sitosterol,kaempferol,stigmasterol,isorhamnetin,etc;There are 186 intersection targets of Baoyuan Tang and AS,and its core targets include AKT1,JUN,TNF,TP53,STAT3,IL6,etc;The GO function enrichment analysis results suggest that it mainly involves 405 biological processes,50 cell components and 78 molecular functions.It mainly participates in biological processes such as positive regulation,signal transduction,apoptosis and inflammatory reaction through cell components such as cytoplasm,nucleus and cytoplasm,and exerts molecular functions such as protein binding,identical protein binding and enzyme binding;The results of KEGG pathway enrichment suggest that Mingjunsan plays an anti-AS role mainly through multiple signal pathways such as lipid and AS,AGE-RAGE pathway,MAPK pathway,PI3K/AKT pathway,Toll-like receptor pathway and so on.3.Experimental study:(1)The HE staining results showed that compared with the blank group,the thickness of the aortic vascular wall of the model group rats was uneven,the structure was disordered,the intima was significantly thickened,the endothelial cells were shed,the endothelium was incomplete,a large number of foam cells were formed,inflammatory cells were infiltrated,the smooth muscle cells of the middle membrane were proliferated and arranged disorderly,the elastic fiber layer was broken,forming typical atherosclerotic plaques,and the lumen was narrowed.The pathological changes of AS in each intervention group were improved to varying degrees compared with the model group.The number of foam cells under the intima was reduced,the degree of inflammatory cell infiltration was reduced,and smooth muscle cells were slightly proliferated and arranged regularly.(2)Blood lipid level: compared with the model group,the serum TC,TG,LDL-C levels in the western medicine group and the middle and high dose groups of Mingjun powder were significantly lower,while the HDL-C levels were significantly higher(P<0.05 or 0.01).(3)Inflammatory factor level: compared with the model group,serum IL-6 and IL-1β 、TNF-α of rats in the western medicine group and Mingjun powder groups were significantly lower(P<0.01).(4)Endothelial factor level: compared with the model group,the serum ET-1 level of rats in the western medicine group and Mingjunsan groups were significantly decreased,and the NO level was significantly increased(P<0.01).(5)RT-PCR results showed that compared with the blank group,the m RNA expression level of TLR4,MyD88,NF-κB in the aortic tissue of rats in the model group was significantly increased(P<0.01);Compared with the model group,The m RNA expression level of TLR4,MyD88,NF-κB in the aorta tissue of rats in the western medicine group and Mingjun powder groups decreased(P<0.01).(6)Western-blot results showed that compared with the blank group,The expression protein level of TLR4,MyD88,NF-κB in the aortic tissue of rats in the model group increased significantly(P<0.01);Compared with the model group,the expression protein level of TLR4,MyD88,NF-κB in the aorta tissue of rats in the western medicine group and Mingjun powder groups decreased(P<0.05 or 0.01).Conclusion:1.Clinical study: Mingjun San is effective in the treatment of stable angina pectoris with yin-cold stagnation evidence,and can effectively improve the symptoms of angina pectoris,improve the survival quality of patients,improve the efficacy of electrocardiogram,increase the rate of stopping and reducing nitroglycerin,improve the efficacy of Chinese medicine evidence,reduce the level of blood lipids,reduce the level of inflammation index hs-CRP,and there are no obvious toxic side effects and adverse reactions during the study,and the safety is high.2.Network pharmacology: Mingjunsan contains 124 active ingredients and 221 related targets in total.Its active ingredients may play a role in the process of positive regulation,signal transduction,apoptosis,inflammatory reaction and other processes by acting on multiple signal pathways,which preliminarily reveals the potential mechanism of anti-AS action of Mingjunsan with multiple components,multiple targets and multiple pathways.3.Experimental study: Mingjunshan can effectively improve the serum lipid level in rats,reduce the levels of serum inflammatory factors TNF-α,IL-1β and IL-6 in rats,balance the serum NO/ET-1 levels,and inhibit the expression levels of TLR4,MyD88,NF-κB m RNA and their proteins in the aortic tissue of AS rats,thereby exerting anti-AS effects,and the mechanism may be related to its inhibition of the inflammatory response mediated by the TLR4/MyD88/NF-κB signaling pathway. |
PDF Full Text Request