| Objective The animal myocardial infarction model was used to verify the curative effect of fistular onion stalk extract on prevention and treatment of ventricular arrhythmias after myocardial infarction.The effect of drugs on myocardial-related ion channels was verified by isolating cardiomyocytes,and the mechanism of Fistular onion stalk extract was initially elucidated.RNA and combined with bioinformatics prediction of target protein,further clarify the onion extract treatment of myocardial infarction after ventricular arrhythmia micro RNA regulatory mechanisms.Method In this study,SD male rats weighing 250-300 g and 3-4 months of age were randomly divided into a preventive group and a treatment group.The prevention group(AL)was divided into: sham operation group,blank model group,light blue group,light blue extract group and lidocaine group,15 in each group;treatment components were acute group(1 week)and chronic group(4 weeks),Each group was divided into sham operation group,blank model group,light blue group,light blue extract group and lidocaine group,15 in each group.Intraperitoneal injection of anesthesia,upper ventilator,thoracotomy,exposure of the heart,silk thread around the left anterior descending branch ligation,until the myocardium in the range of significant swelling,darkening,no contraction movement,ECG monitoring ST segment elevation 0.5 m V,and Within 30 minutes,the myocardial infarction model was successfully prepared.In the prevention group,the body surface electrocardiogram was connected after anesthesia.After the model was successfully established,the stimulating electrodes were set up,and ventricular arrhythmia was induced by programmed electrical stimulation.After the administration,the rats were anesthetized and connected to the surface electrocardiogram and opened the chest.,Stimulate the electrodes,and perform electrical stimulation to induce ventricular arrhythmias.In the prevention group,the myocardial infarction model was established after continuous administration for 1 week.After the establishment of the myocardial infarction model in the treatment group,the administration was performed by intragastric administration from the second day after the operation.The acute group received continuous administration for 1 week,and the chronic group received continuous administration for 4 week.At the end of treatment,arrhythmias were induced in vivo and the types of ventricular arrhythmias that occurred were recorded.Arrhythmia scores were compared between groups and efficacy evaluations were performed.Micro RNA microarray was used to compare the differential expression of mi Rs in myocardial cells of 20 SD rats in the prevention and treatment groups,and the original expression profile was restored after drug treatment.List reversible differences in mi Rs expression profiles.The corresponding primers were designed and QRT-PCR was used to further verify the above reversibly differentially expressed mi Rs.Target Scan and Pic Tar tools were used to predict mi Rs target protein.According to protein function,S.oleifera extract-mi RNA-m RNA protein pathway was established.The sequence specificity of mi Rs and target protein was verified in HEK293 cells by luciferase assay,and the corresponding mi Rs were up-regulated and/or down-regulated.,Detect the correlation of protein expression in cardiomyocytes.Result 1.In the prophylactic administration experiment,compared with the blank model group,there was no significant difference in the occurrence of VT and VF between the VP,VT and lidocaine groups in the light blue group(P>0.05).The rest of the treatment groups could be reduced.The number of arrhythmias occurred in rats after myocardial infarction(P<0.01),and the effect of the onion white extract group was the most significant(P<0.01).Compared with the blank model group,there was no significant difference in the occurrence time,duration and occurrence rate of arrhythmia in the light blue group(P>0.05).The light blueberry extract group and lidocaine group could prolong the arrhythmia occurrence time and reduce the duration.The time and incidence(P<0.01)was the most significant in the lidocaine group(P<0.01).Compared with the blank model group,there was no significant difference in the ST elevation and the Mest score between the light blue group and the white light group(P>0.05).Both the light blue and lidocaine groups could reduce the ST segment elevation and the Mest score.<0.05),the light blue extract group had the most significant effect(P<0.01).Compared with the blank model group,there was no significant difference in the number of occurrences of VT and VF in the acute whitening group and the lidocaine group(P>0.05).The rest of the treatment groups could reduce the arrhythmia in the rat after myocardial infarction.The number of occurrence cases(P<0.01)was the most significant(P<0.01).Compared with the blank model group,there was no significant difference in the occurrence time,duration and occurrence rate of arrhythmia in the light blue group(P>0.05).The light blueberry extract group and lidocaine group could prolong the time of arrhythmia and reduce the duration of arrhythmia.The time and incidence(P<0.01)were the most significant(P<0.01).Compared with the blank model group,there was no significant difference in the occurrence time,duration and occurrence rate of arrhythmia in the light blue group(P>0.05).The light blueberry extract group and lidocaine group could prolong the time of arrhythmia and reduce the duration of arrhythmia.Time and incidence (P<0.01).In the chronic administration experiment,compared with the blank model group,there was no significant difference in the number of VP and VF occurrences in the light white group and the lidocaine group(P>0.05).The rest of the treatment groups could reduce the arrhythmia in the rat after myocardial infarction.The number of occurrence cases(P<0.01)was the most significant in the onion white extract group(P<0.01).Compared with the blank model group,there was no significant difference in the occurrence time,duration and occurrence rate of arrhythmia in the light blue group(P>0.05).The light blueberry extract group and lidocaine group could prolong the time of arrhythmia and reduce the duration of arrhythmia.The time and incidence(P<0.01)were the most significant(P<0.01).Compared with the blank model group,there was no significant difference in the occurrence time,duration and occurrence rate of arrhythmia in the light blue group(P>0.05).The light blueberry extract group and lidocaine group could prolong the time of arrhythmia and reduce the duration of arrhythmia.Time and incidence(P<0.01).2.Using mi RNA microarrays,we listed mi RNAs expression profiles in rat cardiomyocytes after treatment of myocardial infarcts with 10 onion white extracts and non-irrigated white extracts in control rats.The scatter plot can be used for the visual evaluation of different expression of mi RNAs in the Fistular onion stalk extract treatment group and the S.oleifera extract treatment group.The X-axis and Y-axis values in the graph represent the normalized average(log2 scale)of the two sets of sample signals,and the green line is the fold change line.The green line at the center of the graph represents a fold change of 1 fold,indicating no difference between the two groups of samples.The mi RNAs located above the green line above and below the green line at the bottom indicated that the fold change between the two groups of samples was greater than 1.5 fold.Scatter plots are used to screen out general data with a fold change of 1.5 or more.The volcano plots can be used to show the relationship between fold change and statistical significance,and it is more intuitive to show a significant difference in the expression of mi RNAs between the Fistular onion stalk extract treatment group and the S.oleifera extract treatment group.The two vertical green lines indicate up-or down-regulation of 1.5-fold,the horizontal green line indicates a P-value of 0.05,the red marker indicates a difference of 1.5-fold or more,and the P-value is less than 0.05 with statistically significant mi RNAs.mi RNAs were used to screen 67 remarkably differentially expressed mi RNAs in reversibly regenerated myocardium of rats treated with Fistular onion stalk extract(FC>1.5,P<0.05).Among them,16 mi RNAs were up-regulated and 51 mi RNAs were down-regulated.The most obvious mi R-21 was up-regulated and the most obvious mi R-15b-5p was down-regulated as a subsequent validation subject.In the q RT-PCR validation of 15 groups of samples,mi R-15b-5p expression was significantly reduced(p<0.05)in the myocardium of the S.oleifera extract group compared to the control group without the S.oleracea extract(p<0.05).The expression of 21 was significantly increased(p<0.05).The expression levels of mi R-15b-5p and mi R-21 were consistent with their expression patterns in mi RNAs.3.Based on the Target Scan database(version 7.0),the target genes of WN3 A and mi R-21,which are mi R-15b-5p related to arrhythmia,were identified(Fig.3.1).In HEK293 cells,mi R-15b-5p and mi R-21 were overexpressed,respectively,and their inhibition of the reporter gene containing the 3’UTR of the target gene was examined.Among them,mi R-15b-5p significantly inhibited the expression of the reporter gene containing the 3′UTR of wild-type(WT)WNT3A gene(P<0.01),while the expression of the 3′UTR of the mutant(Mut)WNT3A gene showed no obvious expression.Differences;Similarly,mi R-21 significantly inhibited the expression of the reporter gene containing the 3′ UTR of the wild-type(WT)TGFβR2 gene(P<0.01),whereas the reporter gene expression of the 3′ UTR of the mutant(Mut)TGFβR2 gene was absent.Significant differences.After knockdown of mi R-15b-5p and overexpression of mi R-21 in rat myocardial cells after myocardial infarction,compared with the control group,mi R-15b-5p knockdown target protein WNT3 A expression levels increased,mi R-21 over The expression level of the target protein TGFβR2 decreased in the expression group.Conclusion 1.Xin Wen Tong Yang traditional Chinese medicine onion extract can improve the incidence of ventricular arrhythmias after myocardial infarction by affecting the type,occurrence and duration of ventricular arrhythmias,and the degree of ST elevation in rat models of myocardial infarction.Development occurs.2.The mechanism of micro RNA inhibition of ventricular arrhythmias after acute myocardial infarction by onion white extract is related to the regulation of expression of mi R-15b-5p and mi R-21,which may affect Wnt/β-catenin and TGF-β/Smad signaling.The transduction pathways are relevant and their specific regulatory mechanisms need to be further studied. |
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