Study On The Mechanism Of Xiaoxianxiong Jiawei Decoction In The Treatment Of Coronary Heart Disease (Phlegm Heat Syndrome) Based On Differential Expression Of LncRNA TUG1

Objective:Coronary atherosclerotic heart disease(CHD)is a heart disease caused by coronary artery atherosclerosis,which leads to stenosis or occlusion of the vascular cavity,resulting in myocardial ischemia,hypoxia or necrosis.A report compiled by the National Center for cardiovascular disease,which mainly reflects the prevalence and prevention of cardiovascular diseases in China,the analysis of China cardiovascular disease report 2018 indicates that:at present,300 million people in China have been diagnosed with cardiovascular disease,of which the number of patients with coronary atherosclerotic heart disease is more than 11 million,ranking second in cardiovascular disease,and the mortality rate is in cardiovascular disease The top of the list is a huge burden on society and families.Therefore,safe and effective prevention and treatment work is of great significance to reduce the mortality of coronary heart disease.Endothelial dysfunction plays an important role in the occurrence and development of coronary heart disease,which runs through the whole process of the initiation,development and clinical events of coronary atherosclerosis.As a treasure of Chinese nation,traditional Chinese medicine has a unique advantage in protecting vascular endothelial and improving vascular endothelial function and preventing coronary heart disease after time precipitation and repeated verification.In recent years,with the continuous innovation and progress of high-throughput gene sequencing technology and gene transcriptomics,nc RNA has opened the mystery in various difficult diseases.The research on nc RNA in cardiovascular diseases is not rare,which provides important ideas for the prevention and treatment of cardiovascular diseases and creates a foundation for gene therapy of diseases.Taurine up regulated 1(TUG1)was first expressed in the cultured retinal cells of newborn mice in vitro,and later proved to be highly expressed in the injured endothelial cells.It can participate in the pathogenesis of cardiovascular disease by mediating the injury of vascular endothelial cells.P38 mitogen activated protein kinases(p38MAPK)signaling pathway plays an important role in endothelial cell injury.It can induce transcription of various related genes through phosphorylated transcription factors,regulate the expression of active substances,and participate in the oxidative stress injury and apoptosis of endothelial cells.Based on the differential expression of LncRNATUG1 and p38 MAPK signaling pathway in the vascular endothelial injury of coronary heart disease,this study will take the protection of vascular endothelial cells as the breakthrough point to further the mechanism of xiaojiexiong Jiawei Decoction in the treatment of coronary heart disease,in order to provide scientific basis for the clinical application of xiaojixiong Jiawei decoction.Method:1.Through theoretical research to explore the name of coronary heart disease,etiology and pathogenesis,the historical origin and progress of traditional Chinese medicine treatment,reveal the TCM dialectical thinking of Xiaoxianxiong Decoction in the treatment of coronary heart disease(phlegm heat syndrome).Based on the unique advantages of traditional Chinese medicine in improving vascular endothelial function in the treatment of coronary heart disease,the potential mechanism of Xiaoxianxiong Decoction in the treatment of coronary heart disease(phlegm heat syndrome)was explored from the level of gene and molecular biology,so as to lay a solid theoretical foundation for further research.2.Clinical observation of Xiaoxianxiong Jiawei Decoction in the treatment of coronary heart disease(phlegm heat syndrome)and the influence of vascular endothelial function.Methods: 56 patients with coronary heart disease(phlegm heat syndrome)were randomly divided into Xiaoxianxiong Jiawei Decoction group and control group.Xiaoxianxiong Jiawei Decoction group was given Xiaoxianxiong Jiawei Decoction on the basis of conventional western medicine treatment in the control group.The course of treatment was 30 days.(1)clinical indicators: clinical symptoms of traditional Chinese medicine,attack times,duration and attack degree of angina pectoris;(2)laboratory indicators: ECG examination records The changes of S-T segment and T wave during the attack of angina pectoris;the secretion levels of nitric oxide(NO)and endothelin(ET)related indexes of vascular endothelium;(3)the evaluation indexes of adverse drug reactions,such as routine blood test,routine urine test,liver and kidney function,etc.3.Study on the relationship between LncRNA TUG1 and vascular endothelial injury in patients with coronary heart disease(phlegm heat syndrome).Randomly selected 15 cases of coronary heart disease(phlegm heat syndrome)patients,another 15 cases of healthy control group.Real time PCR was used to detect LncRNA TUG1 in peripheral blood mononuclear cells.The number of circulating endothelial cells was measured by immunological method.The level of nitric oxide secretion was measured by nitrate reductase method.The level of endothelin secretion was measured by radioimmunoassay.Pearson correlation analysis was used to analyze the correlation between LncRNA TUG1 and circulating endothelial cell count,nitric oxide and endothelin.4.Based on the study of serum pharmacology and time effect relationship of traditional Chinese medicine,explore the mechanism of Xiaoxianxiong Decoction in regulating differential expression of LncRNA TUG1 and p38 MAPK signaling pathway to protect vascular endothelial cells.The serum of Xiaoxianxiong Jiawei Decoction and blank control serum were prepared by intragastric administration of SPF grade SD rats;the endothelial injury model of human umbilical vein endothelial cells(HUVEC)induced by TNF-α was prepared,and the optimal concentration and action time of serum containing Xiaoxianxiong Jiawei Decoction were screened by MTT method.The experiment was divided into model group,blank serum group and Xiaoxianxiong Jiawei decoction containing serum group.The medium,blank group serum and the best Xiaoxianxiong Jiawei decoction containing serum were used for intervention.RT-PCR was used to detect the expression of LncRNA TUG1 in TNF-α induced injured HUVEC,and Western blot was used to detect the expression levels of p38 MAPK signaling pathway related proteins p38 and P-p38 in TNF-αinduced injured HUVEC.Pearson correlation analysis was used to analyze the correlation between differential expression of LncRNA TUG1 and p38、P-p38.Results:1.xiaoxianxiong Decoction originated from the treatise on febrile diseases.Later doctors have been expanding their meaning on the basis of the original text,and clearly proposed that xiaoxianxiong decoction can improve the treatment of coronary heart disease by vascular endothelial from multiple targets,multiple ways and levels under the condition of examining the pathogenesis of phlegm heat junction.2.the clinical effect of xiaoxianxiong Jiawei Decoction on coronary heart disease(phlegm heat syndrome)and its effect on vascular endothelial function.The difference between xiaoxianxiongjiawei Decoction group and the control group was statistically significant(P < 0.05),the total effective rate was92.86% and 96.43% respectively,the total effective rate of ECG was92.86%,which was better than that of the control group(P > 0.05).After treatment,the NO secretion level was up regulated(P < 0.05)and et secretion level decreased(P < 0.05).There was no clinical significance in the two groups,such as blood routine,urine routine,liver and kidney function before and after treatment.One case of stomach pain(3.57%)and 2 cases of abdominal distention(7.14%)appeared in xiaojiexiong Jiawei Decoction group.In the control group,2 cases were dizzy and headache(7.14%),2 cases had abdominal distension(7.14%).3.the relationship between LncRNA TUG1 and vascular endothelial injury in patients with coronary heart disease(phlegm heat syndrome).The relative expression of LncRNA TUG1 in peripheral blood of patients with coronary heart disease(phlegm heat syndrome)increased;The difference between the two indexes was statistically significant(P < 0.05),which were the increase of the number of endothelial cells,the increase of ET secretion and the decrease of NO secretion.Pearson correlation analysis showed that LncRNA TUG1 in peripheral blood of patients with coronary heart disease(phlegm heat syndrome)was positively correlated with the number of circulating endothelial cells(r=0.56,P=0.03),and negatively correlated with the level of NO secretion(r=-0.58,P=0.02),and also positively correlated with ET secretion level(r=0.62,P=0.01).4.the mechanism of regulating the differential expression of LncRNA TUG1 and p38 MAPK signaling pathway in xiaoxianxiong Jiawei Decoction protects vascular endothelial cells.The time-dependent study found that the serum of xiaoxianxiong jiawei Decoction improved TNF in a time dose-dependent manner-α The ability of inducing the value-added of HUVEC cells was induced.Finally,20%xiaoxianxiong Jiawei decoction was used to intervene TNF-α HUVEC cells were injured for 24 hours.The results of real time PCR showed that the average expression level of LncRNA TUG1 was 1.00 in the model group,0.98 in the blank serum group,0.96 in the 20%xiaoxianxiong jiawei Decoction group,and the difference was statistically significant(P < 0.05).The Western blot results showed that the expression of p38 and P-p38 in 20% xiaoxianxiong jiawei decoction was significantly lower than that in the model group and blank serum group(P < 0.05).Pearson correlation analysis showed that the expression of LncRNA TUG1 was positively correlated with the expression of p38 and p-p38,respectively(r=0.997,P=0.048),(r=0.990,P=0.017).Conclusion:1.xiaoxianxiong Jiawei decoction combined with western medicine can effectively improve the clinical symptoms and vascular endothelial function of coronary heart disease(phlegm heat syndrome).2.there are differences in the expression of LncRNA TUG1 in peripheral blood of patients with coronary heart disease(phlegm heat syndrome),and there is a certain correlation between the differential expression of LncRNA TUG1 and vascular endothelial injury.3.the serum of xiaoxianxiong jiawei decoction can reduce the expression of p38 MAPK signal pathway related protein by regulating the differential expression of LncRNA TUG1,and reduce endothelial cell damage.4.xiaoxianxiong Jiawei decoction can improve the treatment of coronary heart disease(phlegm heat syndrome)by vascular endothelial treatment from gene level and molecular biology level on the premise of examining the pathogenesis.