| Purpose:Atrial fibrillation can induce complications such as cardiac insufficiency and cerebral embolism.It has a high disability rate and mortality rate.It is a difficult clinical cardiovascular disease.Exploring its pathological mechanism and seeking clinical effective treatment methods that can benefit patients for a long time have increasingly become a research hotspot in the medical field.This study designed two parts of clinical observation and animal experiment around the traditional Chinese medicine compound for removing phlegm and removing blood stasis to observe the clinical effectiveness of the traditional Chinese medicine compound for removing phlegm and removing blood stasis and explore its possible mechanism,so as to provide a reliable scientific basis for its clinical application.Material and method:1.Observation on the clinical efficacy of traditional Chinese medicine compound for removing phlegm and removing blood stasis in the treatment of paroxysmal atrial fibrillation(phlegm turbidity and blood stasis type): 110 patients with paroxysmal atrial fibrillation of phlegm turbidity and blood stasis type were randomly divided into control group and treatment group,with 55 cases in each group.The patients in the control group took amiodarone hydrochloride tablets orally,and the treatment group took traditional Chinese medicine compound granules for removing phlegm and removing blood stasis orally for 12 weeks as a course of treatment.Those with other basic diseases continued to take conventional western medicine orally.The curative effect of TCM syndrome,score of main TCM symptoms,duration of atrial fibrillation,attack frequency and recovery maintenance rate of three months before and after treatment were compared between the two groups.2.Effects of Chinese herbal compound for removing phlegm and removing blood stasis on oxidative stress and sympathetic nerve function in atrial fibrillation model rats: 90 male SD rats were randomly divided into two groups: blank group(n=15)and model group(n=75).The rat model of atrial fibrillation was prepared by injecting Ach(66ug/ml)-Ca Cl2(10mg/ml)mixture through the tail vein of rats.On the 7th day of modeling,all rats were weighed,10%chloral hydrate was anesthetized,connected to a Power Lab 15 T multi-conductivity physiological recorder,and the electrocardiogram of lead II was measured.Irregular f waves and absolute unequal R-R intervals are a successful model for atrial fibrillation.The 75 successfully modeled rats were randomly divided into model group,Chinese medicine high,medium and low dose groups and verapamil group,with 15 rats in each group.On the 2nd day after the successful model establishment,the groups were given Chinese herbal compound 41.25,20.63,and 10.31 g/(kg·d)by gavage,respectively,and the verapamil group was given 25 mg of verapamil solution./(kg·d)was administered by intragastric administration for 2 consecutive weeks,and the tail vein injection was continued during the period,which was performed 1 hour after intragastric administration.After 2 weeks,the electrocardiogram of lead II of the rats was measured again,and the duration of atrial fibrillation was calculated;HE staining was used to observe the pathological morphology of the atrial myocardium of the rats;transmission electron microscopy was to observe the ultrastructure of the atrial myocytes in each group;The content of ROS,SOD,GSH-Px and NPY in mouse serum.3.Based on CAV1.2 / CAM / CaMKⅡpathway to explore the effect of traditional Chinese medicine compound for removing phlegm and removing blood stasis on calcium regulation in atrial fibrillation model rats: the experimental steps are the same as above.RT-PCR was used to detect cav1 in atrial muscle tissue CAV1.2、Ca M、Ca MKⅡ、FKBP12.Relative expression of 6 m RNA;Cav1 was measured by Western blot CAV1.2、Ca M、Ca MKⅡ、Ry R2、P-Ry R2、FKBP12.6 protein expression level: cav1 in atrial muscle tissue was detected by immunohistochemistry CAV1.2、Ca M、Ca MKⅡ、Ry R2、P-Ry R2、FKBP12.6 protein expression.Graphpad Prism version 7.0 was used for statistical analysis and data analysis charts,and the measurement data were expressed as x±S.The ANOVA variance test was used for comparison between groups.When P<0.05,the difference was considered to be statistically significant,and P<0.01 was considered to be significantly different.Results:1.Observation of clinical efficacy of Chinese herbal compound for removing phlegm and removing blood stasis in the treatment of paroxysmal atrial fibrillation(phlegm turbidity and blood stasis type):1.1 Comparison of the efficacy of traditional Chinese medicine syndrome: the total effective rate of the treatment group was 86%(43/50),and the total effective rate of the control group was 46%(23/50).P < 0.05).1.2 Comparison of the main TCM symptom scores: The main TCM symptom scores of the two groups of patients after treatment were lower than those before treatment,and the TCM symptoms in the treatment group were improved more significantly than those in the control group(P<0.05).1.3 Comparison of the frequency of atrial fibrillation(times/week): the frequency of atrial fibrillation in the two groups after treatment was significantly lower than that before treatment(P<0.01),and the duration of the treatment group was more significantly lower than that in the control group(P<0.05).1.4 Comparison of the duration of atrial fibrillation(h): the duration of atrial fibrillation in the two groups after treatment was significantly lower than that before treatment(P < 0.01),and the frequency of attacks in the treatment group was more significantly lower than that in the control group(P < 0.05).1.5 Comparison of recovery and maintenance rates for three months of treatment: The three-month recovery and maintenance rate of patients in the treatment group was 80%,and that in the control group was 38%,and the treatment group was better than the control group(P<0.01).2.The effect of Chinese herbal compound for removing phlegm and removing blood stasis on oxidative stress and sympathetic nerve function in rats with atrial fibrillation:2.1 The general state and survival number of rats in each group: Except for the blank group,all rats have different degrees of weight loss,mental fatigue,lack of luster,and different lodging.After the tail vein injection,the rats in the model group had increased breathing and heart rate,congested eyeballs and even oozing blood,and recovered within 10-15 minutes,showing weakness,drowsiness,lying down,loss of appetite,and weight loss;the rats in the high and medium dose groups of traditional Chinese medicine and the verapamil group Compared with the model group,the oozing blood in the eyeball was reduced,the activity was relatively high,and there was no obvious loss of appetite.After the experiment,a total of 74 rats remained,10 died due to intestinal obstruction after autopsy,2 died due to inhalation into the lungs by gavage,and 4 died due to excessive tail vein injection.2.2 Comparison of the duration of ECG atrial fibrillation in each group: compared with the blank group,the rats in the model group had typical atrial fibrillation(P<0.01);Compared with the verapamil group,the duration of atrial fibrillation in the middle-dose Chinese medicine group had no significant difference(P>0.05).2.3 Pathological changes of atrial myocardium of rats in each group: The atrial myocardium of rats in the blank group was evenly stained,the cells were normal in shape,closely arranged,and striated neatly.The myocardial cells in the model group were swollen,the size of the nucleus was different,the fibrous tissue was disordered and broken,filled with collagen fibers,the gap was widened,the arrangement was loose,and the eosinophilic granules increased.Compared with the model group,the myocardial cell swelling was improved,the intercellular space was narrowed,and the myocardial fibers were arranged relatively neatly in the high and medium dose groups of traditional Chinese medicine and the verapamil group.2.4 The ultrastructural changes of atrial myocytes in each group: the sarcomere of the atrial myocytes of the rats in the blank group were complete and clear,the "Z" band was neatly arranged and coherent,the I-band was clearly visible,the myofilaments were neatly and tightly arranged,the mitochondrial structure was normal,and the mitochondrial cristae were The structure is complete.The sarcomeres of the atrial myocytes of the rats in the model group were broken,the "Z" band was discontinuous,the arrangement was disordered,the "I" band was indistinct,the myofilaments of the cardiomyocytes were dissolved and broken,voids appeared,mitochondria were swollen,increased,and even ruptured,and the cristae was broken.Dissolved,aggregated together,and vacuolated.The sarcomere of the atrial myocytes in each dose group of traditional Chinese medicine and verapamil group were slightly broken,and the arrangement was relatively neat.2.5 Changes of serum ROS,SOD and GSH-Px contents in each group: Compared with the blank group,the serum ROS contents of the model group were increased,while the GSH-Px and SOD contents were significantly decreased(P<0.01);compared with the model group,the content of serum ROS decreased and the content of SOD increased in the high and medium dose groups of traditional Chinese medicine and verapamil group(P<0.05);the content of serum GSH-Px in each dose group of traditional Chinese medicine and verapamil group was significantly increased(P<0.01).);compared with the verapamil group,there was no significant difference in the contents of ROS,GSH-Px and SOD in the middle-dose Chinese medicine group(P>0.05).2.6 Changes of serum NPY content of rats in each group: Compared with the blank group,the serum NPY content of rats in the model group was significantly increased(P<0.01);Compared with the verapamil group,there was no significant difference in the content of NPY in the middle-dose group of traditional Chinese medicine(P>0.05).3.Based on the CAV1.2/CaM/CaMKⅡ pathway to explore the effect of the traditional Chinese medicine compound for removing phlegm and removing blood stasis on calcium regulation in atrial fibrillation model rats:3.1 Comparison of the relative expression levels of CAV1.2,Ca M,Ca MKⅡ,FKBP12.6m RNA in the atrial muscle tissue of the rats in each group: Compared with the blank group,the relative expression levels of CAV1.2 and FKBP12.6 m RNA in the atrial muscle tissue of the rats in the model group decreased,while The relative expressions of Ca M and Ca MKⅡm RNA increased(P<0.05);compared with the model group,the relative expressions of CAV1.2 and FKBP12.6 m RNA in each dose group of traditional Chinese medicine and verapamil group were increased(P<0.05).The relative expression of m RNA decreased;compared with the verapamil group,the relative expression of CAV1.2 m RNA in the middle-dose Chinese medicine group was higher than that in the verapamil group(P<0.05);There was no difference between the relative expression and the verapamil group(P>0.05).3.2 Comparison of CAV1.2,CaM,CaMKⅡ,RyR2,P-RyR2,FKBP12.6 protein expression in atrial muscle tissue of rats in each group: Compared with blank group,the protein expression of CAV1.2 and FKBP12.6 in atrial muscle tissue of model group rats decreased(P<0.01),while the protein expressions of Ca M,Ca MKⅡ,and P-Ry R2 increased(P<0.01),and there was no difference in the expression of Ry R2(P>0.05).The protein expressions of CAV1.2and FKBP12.6 in the Pami group were increased(P<0.01),while the expressions of Ca M,Ca MKⅡ,and P-Ry R2 were decreased(P<0.01),and there was no difference in the expression of Ry R2(P>0.05).Compared with the verapamil group,the protein expression of CAV1.2 in the middle-dose Chinese medicine group was not different from that in the verapamil group(P>0.05);the protein expression of Ca M in the middle-dose Chinese medicine group was higher than that in the verapamil group(P<0.05).The protein expressions of Ca MKⅡ and P-Ry R2 in the middle-dose Chinese medicine group were lower than those in the verapamil group(P<0.05);the protein expression of FKBP12.6 in the middle-dose Chinese medicine group was higher than that in the verapamil group(P<0.01).Conclusion:1.The effective rate of TCM syndromes in the treatment of atrial fibrillation with the Chinese medicine compound for removing phlegm and removing blood stasis reaches 86%,reducing the main TCM symptom scores of patients with atrial fibrillation,reducing the frequency of atrial fibrillation,shortening the duration of atrial fibrillation,and maintaining the recovery for three months.rate of 80%.2.The traditional Chinese medicine compound for removing phlegm and removing blood stasis can shorten the duration of ECG atrial fibrillation in atrial fibrillation model rats,inhibit the changes of pathological morphology of atrial muscle,and reduce the damage of mitochondria and sarcomeres in the ultrastructure of atrial muscle cells.3.The traditional Chinese medicine compound for removing phlegm and removing blood stasis can reduce the expression levels of serum ROS and NPY in atrial fibrillation model rats,and increase the expression levels of serum SOD and GSH-Px in rats,suggesting that the Chinese medicine compound for removing phlegm and removing blood stasis has a protective effect on atrial fibrillation rats.May be related to inhibition of oxidative stress and sympathetic activation.4.The traditional Chinese medicine compound for removing phlegm and removing blood stasis can up-regulate the protein and gene expression of the key target protein CAV1.2 in the CAV1.2/CAM/CAMKⅡ signaling pathway,and down-regulate the protein and gene expression of CAM and CAMKⅡ and their downstream The expression of protein P-RYR2;regulates the balance between RYR2 and P-RYR2,and up-regulates the protein and gene expression of RYR2-stabilizing protein FKBP12.6,thereby inhibiting the disturbance of calcium regulation in atrial myocytes. |
PDF Full Text Request