Study On The Protective Effect And Mechanism Of Lishi Huoxue Recipe On Vascular Endothelial Injury Cell Model Induced By High Uric Acid

ObjectivesNowdays,hyperurice(HUA)is a common disease among the olds,the reason and mechanism of HUA are complicated,symptoms are various,and it's attack is not obvious.Clinical symptoms is nontypical.It can lead to all kinds of cardiovascular disease and chronic kidney disease.The trend of attack is getting younger in recent years.Up to now,etiological factor and Pathogenesis of HUA are complex.This project's starting point is uric acid,which effected the vascular endothelium function,and cause serious and extensive harm to body.To protect the endothelial function in the process of treatment as the main line.Used the theory of traditional Chinese medicine,and explore the root to disease.The treatment for HUA by adjusting the circulating blood,Yin-yang balance ofhumanbody,regarding the characteristics of primary asthenia-secondary stheniasyndrome,enhances resistibility of bodys.The effect and advantages of treatment of HUA by traditional traditional Chinese medicine has been increasingly accepted by inteRNAtional and domestic academics.Western medicine treating HUA apply symptomatic treatment.Lowering uric acid,eliminating the clinical symptoms.But the side effects cannot be ignored.The Chinese herbal compounds of LiShiHuoXue prescription was under the guidance of traditional Chinese medical theory,Combines with the clinical experience of doctor,LSHXF is very effective in clearing away heat and dampness and promoting circulation of blood and removing stasis,for the HUA damp-heat pouring downward and blood stasis.This project explore the efficacy and mechanisms of LSHXF,and provides the foundation for drug use rationaly,effectively and safty,which based on the documents and experimental research.MethodsThe experimental study of Lishi huoxue prescription containing serum on vascular endothelial cells by vascular high uric acid treated human umbilical,effects of high concentrations of uric acid on the activity of human umbilical vein endothelial cells,to verify the hyperuricemia cell model,on the basis of the establishment of the blank control group,model group,benzene bromine Malone group,four wonderful pill group and dampness blood low,middle and high dose group and 1 group,2 group and 9 group,respectively,from the cell activity,cell apoptosis,cell adhesion factor,cell contraction factor and inflammatory factor P38-NF-K based on B-MCP-1 path and to detect the corresponding index by CCK-8,AnnxinFITC/PI,ELISA,nitrate reductase method,RT-PCR and WB etc.Results1.the study of the influenceof lishi huoxue prescription on the activity and apoptosis of HUVEC1.1 the study of the influence ofdifferent content of uric acid on the activity of HUVEC1.1.1 different levels of uric acid in HUVEC cells after 24 hours,compared with the control group,when uric acid level is over than or equal to 20 mg/dl,OD value decreased obviously,with significant difference(P<0.01).1.1.2 different levels of uric acid in HUVEC cells after 48 hours,compared with the control group when the level of uric acid is over than 15 mg/dl,OD decreased obviously,with significant difference(P<0.05,P<0.01).1.1.3 different levels of uric acid in HUVEC cells after 72 hours,compared with the control group when uric acid level is over than or equal to 15 mg/dl,OD value dropped significantly,with significant difference(P<0.01).1.2 the study of lishi huoxue prescription effect on the activity of HUVEC treated by high uric acid1.2.1 when the drug-containing serum concentration of 5%,compared with the control group,two groups of high level uric acid in the model group,all cell activity was significantly inhibited(P<0.05);Compared with 20 ua model group,lishihuoxue prescription cell activity rises high dose group,with significant difference(P<0.05);Compared with 40 ua model group,positive medicine Benzbromarone and lishi huoxue prescription cell activity rises high dose group,with significant difference(P<0.05,P<0.01).1.2.2 when the drug-containing serum concentration of 10%,compared with the control group,two groups of high uric acid concentration in the model group,all cell activity was significantly inhibited(P<0.05,P<0.01).Compared with 20 ua model group,lishi huoxue prescription cell activity rises high dose group,with significant difference(P<0.05,P<0.01).Compared with 40 ua model group,lishi huoxue prescription cell activity rises high dose group,with significant difference(P<0.05).1.2.3 when the drug-containing serum concentration of 20%,compared with the control group,two groups of high uric acid concentration in the model group,all cell activity was significantly inhibited(P<0.05).Compared with 20 ua model group,positive medicine Benzbromarone combination lishi huoxue prescription cell activity rises high,medium and low dose group,with significant difference(P<0.05).Compared with 40 ua model group,positive medicine Benzbromarone and lishi huoxue prescription cell activity rises high dose group,with significant difference(P<0.05).1.2.4 The study of the lishi huoxue prescription on influence of HUVEC activity treat by high level uric acid 24 hCompared with the control group,the activity of cell in model group was obviously suppressed(P<0.05).Groups compared with model group,Benzbromarone,Simiaowan group,two level of lishi huoxue prescription groups,in the cell activity significantly raised(P<0.05,P<0.01).1.2.5 The study of the lishihuoxue prescription on influence of HUVEC activity treat by high level uric acid 48 h.Compared with the control group,the activity of cell in model group was obviously suppressed(P<0.05);Compared with model group,benzene bromide and cell activity in lishi huoxue prescription Malone group was obviously raised(P<0.05,P<0.01).1.2.6 The study of the lishi huoxue prescription on influence of HUVEC activity treat by high level uric acid 72 h.Compared with the control group,the activity of cell in model group was obviously suppressed(P<0.05);Groups compared with model group,Benzbromarone group,lishihuoxue prescription group cell activity lower(P<0.05,P<0.01).1.3 the effect of lishihuoxue prescription on the apoptosis of HUVEC treated by high level uric acid1.3.1 the effect of lishihuoxue prescription on the apoptosis of HUVEC treated by high level uric acid:the influence of apoptosis rate compared with the control group,model group significantly increased(P<0.001).Compared with model group,Benzbromarone,lishi huoxue prescription high dose group and the apoptosis rate was significantly lower in the 2 groups(P<0.001,P<0.01,P<0.05),Simiaowan group and party 1 has a downward trend but no statistical significance(P>0.05),lishi huoxue prescription compared two groups,the dose effect is better than that of high dose;Compared with the dose in lishi huoxue prescription group,there are differences between the Simiaowan group(P<0.05).The results are shown in table 61.3.2 the effect of lishihuoxue prescription on the apoptosis of HUVEC treated by high level uric acid:Compared with the control group,model group,apoptosis rate increase significantly(P<0.001).Compared with model group,the apoptosis rate of drug groups were obviously lower(P<0.001,P<0.01,P<0.05);Compared with the dose in lishi huoxue prescription group,the effect of high dose group on the apoptosis rate is more obvious.2.the study of the effects of lishihuoxue prescription on the level of cell adhesion molecules of HUVEC treated by high level uric acid2.1 the effect of lishi huoxue prescription on HUVEC treated by high level uric acid on the level of sICAM-1.Compared with the control group,after high uric acid stimulation model group on human umbilical vein endothelial cells sICAM-1 content in supeRNAtant increased dramatically,with significant difference(P<0.001).Compared with model group,each dose group of cells that the supeRNAtant fluid sICAM-1 content decreased significantly,with significant difference(P<0.001),of which the positive medicine to reduce the effect is most obvious,lishi huoxue prescription decreasing trend in the dose group of high dose group was obviously,both Decomposed Recipes group decreased obviously in Decomposed Recipes group.Compared with lishi huoxue prescription high dose group,Benzbromarone,Simiaowan group and down side two groups have significant difference(P<0.001).2.2 the study of lishihuoxue prescription on HUVEC treated by high uric acid on the level of ICAM-1 mRNA and VCAM-1 mRNA2.2.1 the change on the level of ICAM-1 mRNA:compared with the control group,model group ICAM-1 mRNA content increased obviously,with significant difference(P<0.001);Compared with model group,levels of ICAM-1 mRNA in dose groups are significantly decreased(P<0.001),and the lishihuoxue prescription group fell the most obvious among dose groups,the whole party two doses group were superior to other dosing,Decomposed Recipes 1 a split party 2 as well.Compared with lishi huoxue prescription high dose group,Benzbromarone group,Simiaowan group and Decomposed Recipes 1,there are differences between the two groups(P<0.001,P<0.05).2.2.2 the change on the level of VCAM-1 mRNA:compared with the control group,model group VCAM-1 mRNA content increased obviously,with significant difference(P<0.001).Compared with model group,VCAM-1 mRNA levels in dosing Groups were significantly decreased(P<0.001),of which the lishi huoxue prescription fell the most obvious high dose group,the whole party two lower VCAM-1 mRNA expression dose group were better than the other group,Decomposed Recipes 2 a split party 1 as well.Compared with lishi huoxue prescription high dose group,Benzbromarone group,Simiaowan group and Decomposed Recipes 1,there are differences between the two groups(P<0.001,P<0.05).2.2.3 the effect oflishi huoxue prescription on the expression of ICAM-1 and VCAM-lprotein of HUVEC treated by high level uric acid3.the influence of lishihuoxue prescription worked on HUVEC treated by high level uric acid on the level of NO,eNOS,ET-13.1 the effect of lishi huoxue prescription worked on HUVEC treated by high level uric acid on the level of NO.Lishi huoxue prescription worked on HUVEC treated by high uric acid secretion on the split party NO content:the influence of the NO content compared with the control group,model group was obviously decreased(P<0.05);Groups compared with model group,the drug has NO content rise in varying degrees,in which Benzbromarone group has a rising trend but NO statistical significance(P>0.05),more than all have significant difference to the drug(P<0.05,P<0.01,P<0.001),of which the lishihuoxue prescription raised the high dose group was obviously.Compared with high dose of lishi huoxue prescription group,Benzbromarone,Simiaowan group and the two groups had significant difference(P<0.01,P<0.001).3.2 the study of lishihuoxue prescription on HUVEC treated by high uric acid on the level of eNOS mRNA and the influence of ET-1mRNA3.2.1 the effects of eNOSmRNA content after high uric acid treatment,compared with the control group,model group eNOSmRNA content significantly decreased,with significant difference(P<0.05).Group compared with model group each drug eNOSmRNA content rising to some extent,in addition to the four wonderful pill group,more than to drug differences were significant(P<0.001),lishi huoxue party all party two dose group difference is not obvious;Compared with lishi huoxue prescription high dose group,Benzbromarone,Simiaowan group and Decomposed Recipes 1,two groups have significant difference(P<0.001).3.2.2 the effects of ET 1 mRNA content after high uric acid treatment,compared with the control group,model group ET-1 mRNA levels increased significantly,with significant difference(P<0.001);Compared with model group,the contents of ET 1 mRNA drugs to some extent,to reduce the differences were significant(P<0.001),of which the lishi huoxue prescription high dose group of lower trend is most obvious;Compared with lishi huoxue prescription high dose group,Benzbromarone,Simiaowan group and Decomposed Recipes 1,with significant difference(P<0.001).3.2.3 lishi huoxue prescription worked on HUVEC treated by high level uric acid on the Decomposed Recipes eNOS and ET-1 protein expression4.the study of lishihuoxue prescription on HUVEC treated by high uric acid on the level of P38 MAPK,NF-?B,COX-2,and MCP-1 gene expression4.1 impact on P38MAPKmRNA content:compared with the control group,model group P38MAPKmRNA content rised significantly(P<0.001);Compared with model group,each dose group P38MAPKmRNA content significantly lowered(P<0.001),of which the lishi huoxue Decomposed Recipes groups to reduce the trend is most obvious,Decomposed Recipes 1 group in the aspect of reducing P38MAPKmRNA content effect is superior to the down side two groups;Compared with lishi huoxue prescription high dose group,Benzbromarone,simiaowan group and the two Decomposed Recipes groups with significant difference(P<0.001,P<0.05).4.2 the impact on the content of NF-?BmRNA:compared with the control group,model group,NF-?B mRNA content increases significantly,the difference was statistically significant(P<0.001).Compared with model group,the content of NF-kBmRNA dosage group was obviously decreased(P<0.001),the lishi huoxue Decomposed Recipes group to reduce the trend is most obvious,Decomposed Recipes group 1 in reducing NF-kBmRNA content effect superior to Decomposed Recipes two groups;Compared with lishi huoxue prescription high dose group,Benzbromarone,Simiaowan group and Decomposed Recipes 1,2groups have significant difference(P<0.001,P<0.05).4.3 the impact on the MCP-1 mRNA content:compared with the control group,model group MCP-1 mRNA level rised significantly(P<0.001);Groups compared with model group,Benzbromarone levels significantly lower MCP-1,the difference was statistically significant(P<0.01),more than a given set of MCP-1 mRNA levels were significantly lower(P<0.001),of which the lishi huoxue party Decomposed Recipes to reduce the trend is most obvious,Decomposed Recipes group in the lower MCP-1 mRNA content effect superior to the Decomposed Recipes 2 group.Compared with lishi huoxue prescription high dose group,Benzbromarone,Simiaowan group and Decomposed Recipes 1,2 groups have significant difference(P<0.001).4.4 the influence on the level of COX-2 mRNA:compared with the control group,model group cox-2 mRNA levels rise significantly(P<0.001).Groups compared with model group,a drug cox-2 mRNA levels significantly lowered(P<0.001),among them the two dose lishihuoxue groups had the best trend,Decomposed Recipes 1 group in lower cox-2 mRNA content effect superior to Decomposed Recipes 2 group;Compared with lishi huoxue prescription high dose group,Benzbromarone,Simiaowan group and Decomposed Recipes 1,2 group have significant difference(P<0.001).ConclusionsTo sum up,this topic through literature review and experimental study,the combination of lishi huoxue prescription function attending doctor of traditional Chinese medicine,the related modern pharmacological experiments,reveals to clear heat,promoting blood circulation to remove blood stasis with wet method of traditional Chinese medicines to create lishi huoxue prescription treatment effectiveness and advantage of the HUAe,discussed mainly been used to eliminate the cause of Decomposed Recipes 1 and to promoting blood circulation to remove blood stasis protection mainly down the side of the body of the 2 in the treatment of the role and its contribution to the overall treatment effect,to elucidate the protective mechanism of animal models with HUA,vascular endothelial,to enhance the level of traditional Chinese medicine in the treatment of HUA theory and experimental foundation for clinical curative effect.