Based On ERK1/2 And TGF-β1/Smad3 Pathways, The Mechanism Of Shenlian Fumai Granules Intervention On Hypertensive Atrial Remodeling And Atrial Fibrillation Susceptibility Was Studied

Atrial fibrillation is a common clinical arrhythmia,which can cause a variety of embolic diseases including stroke,and induce and aggravate heart failure,and seriously threaten human health.At present,the western medicine treatment of atrial fibrillation is mainly based on internal medicine and radiofrequency ablation.Although the curative effect has been significantly improved,there are still certain limitations.Based on the overall concept and syndrome differentiation and treatment of traditional Chinese medicine,it has been found that TCM may be used as a clinical strategy for the prevention and treatment of atrial fibrillation.In clinical practice,it has been found that based on the method of Yiqi and Huoxue,the treatment of atrial fibrillation by adding Qingre and Huatan,Yangxin and Anshen has certain effects.However,due to the uneven sample size,method,and quality level of current clinical research,its effectiveness still lacks accurate judgments.The Meta-analysis,as a powerful means of finding the best evidence for evidence-based medicine,may provide ideas for solving this problem.Therefore,this study intends to adopt the method of Meta-analysis to evaluate the clinical efficacy and safety of Chinese medicines for Yiqi and Huoxue in the treatment of atrial fibrillation.Epidemiology shows that there is a close relationship between atrial fibrillation and hypertension.Hypertensive atrial remodeling is considered to be an important cause of atrial fibrillation.Among them,atrial remodeling with atrial fibrosis as the main feature is the key link.At present,there is still a lack of effective drugs for the remodeling of the atrial structure of hypertension for clinical choice.Research on Chinese medicine may be a breakthrough to solve this problem.The TCM syndromes of hypertension mainly include Ganyang Shangkang,Tanzhuo Zuzhi,qi deficiency and blood stasis,etc.Tachyarrhythmia is based on qi deficiency and blood stasis,and the phlegm-heat syndrome is more prominent,especially in the onset of tachyarrhythmia such as atrial fibrillation,phlegm-heat is particularly obvious.Shenlian Fumai Granule,as an agreed prescription for the treatment of arrhythmia in the cardiology department of our hospital,is based on Yiqi and Huoxue and focuses on Qingre and Huatan,the clinical treatment of atrial fibrillation is effective.Previous studies have found that Shenlian Fumai Granule have the effects of inhibiting inflammation,shortening the duration of action potentials,reducing tachyarrhythmia,and reduce the recurrence rate of atrial fibrillation.Related literature studies have also found that the main components of Shenlian Fumai Granule and active ingredients can inhibit atrial fibrosis and reduce the occurrence of atrial fibrillation by inhibiting the TGF-β1 pathway.In this study,by replicating the animal model of essential hypertension rats,using small animal echocardiography,electrocardiogram,in vivo atrial burst stimulation,electrophysiological examination,immunohistochemistry,and Western Blot methods,from the overall,tissue cell,molecular level conduct research to verify the hypothesis that "Shenlian Fumai Granule may inhibit hypertensive atrial remodeling and atrial fibrillation susceptibility by regulating ERK1/2 and TGF-β1/Smad3 pathways."Objective:1.To explore the clinical efficacy and safety of the treatment of atrial fibrillation with the method of Yiqi and Huoxue,and provide evidence-based medicine for the treatment of atrial fibrillation with traditional Chinese medicine.2.To explore the interventional effects and possible mechanisms of Shenlian Fumai Granule on the remodeling of the atrial of hypertension and the susceptibility to atrial fibrillation.Part Ⅰ Meta-analysis of Chinese medicine for Yiqi and Huoxue in treating atrial fibrillationMethods:Search the Chinese Journal Full-text Database(CNKI),China Biomedical Literature Database(CBM),VIP Chinese Journal Full-text Database(VIP),Wanfang Database through computer search;and foreign databases including PubMed,Web of Science,and Cochrane Library to collect public publications A randomized controlled clinical trial on the treatment of atrial fibrillation with the method of Yiqi and Huoxue combined with western medicine.Meta-analysis of the included literature was performed with RevMan5.3 software,and the results were presented in a forest plot.Sensitivity analysis was used to evaluate whether the results were stable,and the funnel plot was used to analyze whether the literature had publication bias.Result:1.Literature search screening results After literature search and supplementation,a total of 712 documents were retrieved,215 duplicate documents were excluded,the titles and abstracts were browsed,289 documents that did not meet the inclusion criteria were eliminated,and the literature was read through and 17 documents were included.2.Features of included literature2.1 General informationA total of 1639 patients with atrial fibrillation were included in 17 articles,including 828 in the treatment group and 811 in the control group.Two of the 17 articles did not specify the specific age and gender distribution of the treatment group and the control group,and the rest described the treatment in detail the number of cases,gender,age,disease course,intervention measures and course of treatment between the group and the control group.Three of the 17 documents did not clearly describe the course of the disease,and the rest described the course of the disease in detail.And 17 articles all describe the general data of the treatment group and the control group without statistical difference.2.2 Usage of Traditional Chinese MedicineThe literature included in this study are all based on the method of Yiqi and Huoxue,in which the more frequently used drugs are ginseng(10 times),astragalus(6 times),salvia(8 times),and notoginseng(7 times).2.3 Adverse reactions and follow-upAmong the 17 articles included,4 reported no adverse reactions,6 reported adverse reactions,including dizziness,nausea,and vomiting,bradycardia,etc.And 7 did not describe whether adverse reactions occurred.3.The overall quality of the included literature was average.After the modified jadad scoring scale,there were 4 articles with 2 points,6 articles with 3 points,6 articles with 4 points,and 1 article with 6 points.The 17 included papers all mention the word "random",13 of which use the correct random method,none of the 17 documents describe the hiding of the allocation plan,one of the documents uses the correct blinding method,and 2 of the documents report the risk of bias Low,the results of the 17 literature are complete and there is no other source of bias.4.Meta-analysis resultsMeta-analysis results showed that the Chinese medicine for Yiqi and Huoxue combined with conventional western medicine had clinical efficacy(RR=1.16,95%CI=[1.08,1.23],Z=4.33,P<0.00001),and the efficacy of TCM syndromes(RR=1.29,95%CI=[1.18,1.41],Z=4.33,P<0.00001),atrial fibrillation conversion rate(RR=1.23,95%CI=[1.04,1.46],Z=2.46,P=0.01),sinus rhythm Maintenance(RR=1.31,95%CI=[1.14,1.50],Z=3.80,P=0.0001),number of episodes of atrial fibrillation(SMD=-1.00,95%CI=[-1.73,-0.27],Z=2.67,P=0.008)and safety evaluation(RR=0.43,95%CI=[0.28,0.66],Z=3.83,P=0.0001)are better than the conventional western medicine group.Conclusion:Oral Chinese medicine combined with western medicine,mainly based on the method of Yiqi and Huoxue,has certain efficacy and safety in improving the clinical efficacy of atrial fibrillation,the efficacy of TCM syndromes,the conversion rate of atrial fibrillation,the maintenance of sinus rhythm and the number of atrial fibrillation episodes,and has good safety.However,due to the limited quality of the included literature,the results of this meta-analysis have yet to be confirmed by large-sample,high-quality clinical studies.Part Ⅱ Based on ERK1/2 and TGF-β1/Smad3 pathways to study the mechanism of Shenlian Fumai Granule on hypertensive atrial remodeling and atrial fibrillation susceptibilityMethods:1.Grouping and administrationSpontaneously hypertensive rats were randomly divided into SHR blank group(SHR group),SHR+Losartan potassium group(LOS group),SHR+Shenlian Fumai granule group(SLFM group),and normal blood pressure Kyoto West rats as a normal control group(WKY group),there are 4 groups in total.The corresponding drugs or saline were given by gavage for 4 weeks.2.Detection methodAfter 4 weeks of intervention,the rats in each group recorded general conditions such as body weight,blood pressure,heart rate,and death,small animal echocardiography was used to detect the structure and function of the left atrium and left ventricle,the electrocardiogram was used to detect the P wave time limit and PR interval,the multi-channel physiological recorder was used to detect the effective refractory period of the atria,and the atrial burst stimulation was used to detect the susceptibility to atrial fibrillation,the H&E staining,Masson staining and immunohistochemistry to detect atrial cardiomyocyte hypertrophy,atrial fibrosis,the ELISA,Rt-qPCR,and Western Blot technology were used to detect ERK1/2 and TGF-β1/Smad3 pathway-related gene and protein expression levels.Result:1.Model evaluation:(1)Compared with the WKY group,the SHR group was lighter in weight,and both systolic and diastolic blood pressure were significantly increased(P<0.05).(2)Compared with the WKY group,the left ventricular wall increased in the SHR group thickness,reduced left ventricular diameter,enlarged left atrium,decreased left atrial ejection fraction,increased standardized left atrial weight and standardized global weight(P<0.05).(3)Compared with the WKY group,the P wave duration in the SHR group,PR interval,and duration of atrial fibrillation induction increased(P<0.05).(4)Compared with the WKY group,the atrial myocardial cells in the SHR group were hypertrophy,collagen I and collagen III expression increased,and collagen fiber volume Score increased(P<0.05).(5)Compared with the WKY group,the expression of myocardial fibrosis-related factors in the SHR group was abnormal,manifested as the ERK1/2 pathway related factors Ang II,AT1R,and p-ERK1/2 significantly increased(P<0.05),while the TGF-β1/Smad3 pathway related factors Galectin-3,TGF-β1,p-Smad3,MMP-3 increased(P<0.05),and TIMP3 decreased(P<0.05).2.The effects of Shenlian Fumai Granule on the structure and function of the heart:Compared with the SHR group,the SLFM group had a significant reduction in the inner diameter of the left atrium and the maximum volume of the left atrium(P<0.05),and there was no significant difference in other indicators.Compared with the SHR group,the thickness of the left ventricular anterior wall and the inner diameter of the left atrium at the end of diastole were significantly reduced in the LOS group(P<0.05),and the maximum volume of the left atrium tended to shrink,but the difference was not statistically significant,and there was no significant difference in other indicators.3.The effects of Shenlian Fumai Granule on atrial electrophysiology and atrial fibrillation induced rate:Compared with the SHR group,the SLFM group has significantly reduced P wave duration and atrial fibrillation induction duration(P<0.05),and the induction rate of atrial fibrillation tends to decrease,but the difference is not statistically significant.There was no significant difference in other indicators.Compared with the SHR group,the P wave duration,atrial fibrillation induction duration,and PR interval were significantly reduced in the LOS group(P<0.05).Significance,there is no significant difference in other indicators.4.The effects of Shenlian Fumai Granule on atrial cardiomyocyte hypertrophy and atrial fibrosis:Compared with the SHR group,the SLFM group and the LOS group inhibited the atrial cardiomyocyte hypertrophy,the expression of collagen Ⅰ and collagen Ⅲ decreased,and the volume of collagen fibers the score is significantly reduced(P<0.05).5.The effects of Shenlian Fumai Granule on the expression of ERK1/2 and TGF-β1/Smad3 pathway-related factors in atria:Compared with the SHR group,the atrium AT1R and p-ERK1/2/t-ERK1/2 of the SLFM group and the LOS group were significantly reduced(P<0.05).The atrium Galectin-3,TGF-β1,p-Smad3,t-Smad3,MMP-3,MMP-3/TIMP-3 were significantly decreased in the SLFM group and the LOS group(P<0.05),TIMP-3 was significantly increased(P<0.05).Conclusion:1.Shenlian Fumai Granule can improve the atrial structure remodeling in spontaneously hypertensive rats,and have inhibitory effects on atrial electrical remodeling and atrial fibrillation susceptibility.2.The mechanism of Shenlian Fumai Granule in improving atrial remodeling in spontaneously hypertensive rats may be related to the regulation of ERX1/2 and TGF-β1/Smad3 signaling pathways.