The Study About Wumen Antihypertensive Herbal Paste And Its Intervention Of Autophagy To Affect Myocardial Hypertrophy In Hypertension

Objective:Sort out the prescriptions of Doctor Zhao,a prestigious TCM doctor in Wumen by using the Traditional Chinese Medicine Inheritance Support System(TCMISS),summarize the treatment principles,the rules of prescriptions and the characteristics of the medicine used in the treatment of hypertension with herbal paste,so as to better inherit the characteristics of the Wumen herbal paste and guide the clinical practice.Observe the effect of the herbal paste on AngⅡ-induced cardiomyocyte hypertrophy and autophagy at the level of cellular,molecular and protein,then study and explore the effect and mechanism of herbal paste in regulating autophagy to inhibite hypertensive myocardial hypertrophy.Methods:1.Collect and sort out the prescriptions of hypertension patients who treated by Doctor Zhao in the ointment outpatient clinic from January 1,2018 to December 31,2020.Uniformly standardize the name,nature,taste and meridian tropism of the traditional Chinese medicine,then input data into the TCMISS(V2.5)to analyze the frequency and characteristics,summary the data based on association rule analysis and complex system entropy cluster analysis.According to the results,we obtain the clinical experience of Doctor Zhao’s herbal pastes.2.The hypertensive myocardial hypertrophy model was established by AngⅡ-induced H9C2 cardiomyocyte hypertrophy.After treatment with the medicated serum,the cells were randomly divided into six groups:control group(CON),AngII group(ANG Ⅱ),low-herbal paste group(ANG Ⅱ-L),medium-herbal paste group(ANG Ⅱ-M),high-herbal paste group(ANG II-H),irbesartan-contained serum group(ANG Ⅱ-I).Using CCK-8 detected the activity of cardiomyocytes after drug-contained serum treatment.Using Phalloidin staining labeling detected the changes in the surface area of cardiomyocytes in each group.Using real-time PCR detected the expression of ANP、BNP、β-MHC.Using Western Blot detected the relative expression of autophagy protein Beclin1、LC3、p62 and mTOR.Results:1.Clinical data analysis:After analyzing 172 prescriptions by using the TCMISS,the results showed that the drugs contained in the prescription were mostly sweet,warm in nature,and return to the liver and kidney meridian.Based on the analysis of medication mode and association rules,21 drug association rules were obtained.Among them,the drug pairs with higher correlation were gastrodiae-fructus comi,fructus corni-alisma orientalis,fructus corni-amomum,notoginseng-fructus corni,etc.Through complex system entropy cluster analysis,the results show that there were 26 pairs of core drug combinations and 13 new prescriptions.2.Cell experiment results:Compared to control group,the surface area of myocardial cells in the AngⅡ group was significantly increased(P<0.01).Compared to AngⅡgroup,the herbal paste contained serum could inhibit the increase in cell area(P<0.05),and high-herbal paste group was more obvious(P<0.01).Compared to control group,AngⅡ could cause myocardial hypertrophy,increase cell protein content,and up-regulate the expression of target genes ANP、BNP、β-MHC(P<0.05).Compared to AngⅡ group,the herbal paste contained serum could inhibit the increased expression of each gene(P<0.05),and high-herbal paste group was more obvious(P<0.01).Compared to control group,AngⅡ significantly promoted the expression of Beclin1 and LC3-Ⅱ,and inhibited the expression of p62 and mTOR.Compared to AngⅡ group,the herbal paste contained serum could decrease the expression of Beclinl and LC3-Ⅱ and promote the expression of p62 and mTOR(P<0.01),and there were no statistical difference between high-herbal paste group and irbesartan-contained serum group(P>0.05).Conclusion:1.Based on data mining,the results show that the medicine in Doctor Zhao’s herbal pastes are mostly sweet,warm in nature,and return to the liver and kidney meridian.The clinical diagnosis and treatment are based on the principle of "supplement and treatment coexist",which is adept in nourishing the kidney,calming liver,dispersing turbidity and removing blood stasis.The prescription idea advocates "balance is the key",focusing on the balance of yin and yang,harmony of qi and blood,and care of the heart and spleen.Medication advocates "primary and secondary use,dynamic and static combination".It is consistent with the current Wumen ointment formula for the treatment of hypertension,and has its own characteristics.2.According to the experimental results:AngⅡ can induce cardiomyocyte hypertrophy and activate autophagy.After the treatment of herbal paste,the myocardial hypertrophy induced by AngII can be inhibited effectively.The herbal paste can inhibit autophagy of cardiomyocyte hypertrophy by down-regulating the expression of Beclin1 and LC3-Ⅱ,up-regulating the expression of p62 and mTOR.It is speculated that the mechanism of herbal paste in inhibiting hypertensive myocardial hypertrophy may be related to the regulation of autophagy.