Effect Of Danggui Buxue Decoction On Ang? Induced Hypertrophic Cardiomyocytes By TGF-?1/Smad2 And PI3K/Akt Signaling Pathway

Objective:To extract the serum containing Danggui Buxue Decoction and to interfere with the induction of angiotensin II?Ang II?induced hypertrophic cardiomyocytes in vitro,and to explore the effect and mechanism of Chinese medicine Danggui Buxue Decoction on chronic heart failure.Methods:First,the?-actin antibody immunohistochemical method was used to identify cells in accordance with myocardial cells.Secondly,AngII induced cardiomyocytes to establish a model of hypertrophic cardiomyocytes.The expression of ANF mRNA in the cells was measured by BCA method and RT-PCR to verify the model.Microscopic observation of cell morphology of blank group of cardiomyocytes,AngII model group,10%DangGui BuXue Decoction containing serum group and LY294002 blocker group.Determination of ANFmRNA Expression in H9C2 Hypertrophic Myocardial Cells by RT-PCR at Different Concentrations of Serum Containing 20%,15%,10%,5%.Nitric Oxygenase Assay was used to determine the concentration of NO in the culture medium of blank group,AngII model group,10%DangGui BuXue Decoction containing serum group and LY294002 blocker group.Western Blotting was used to measure the expression and activation of TGF?1,Smad2,P-Akt,Akt,P-eNOS,eNOS and other proteins in TGF?1/Smad2 and PI3K/Akt pathways.Result:1.Identification of cell results:Observed by microscope,the cytoplasm contained a lot of brown and fine granular substances,which was positive expression,which was consistent with the characteristics of myocardial cells.2.Modeling results:It was found that the level of ANF mRNA and protein in the AngII model group was higher than that in the normal group?P<0.05?,indicating that the AngII-induced cardiomyocyte hypertrophy model was established successfully.3.Microscopic observation of myocardial cell morphology results:Adherent growth of myocardial cells,its morphology was myocyte-like,while the density and morphology of hypertrophic cardiomyocytes increased compared to normal myocardial cells,floating cells increased.4.Determinationof ANFmRNA expressionincardiomyocytesatdifferent concentrations of drug-containing serum:The expression of ANF mRNA in AngII model group was higher than that in blank group?P<0.05?.There was no significant difference in the expression of ANF between 5%,20%drug-containing serum group and model group?P>0.05?.The expression levels of ANF in the 10%and 15%drug-containing serum groups were lower than those in the model group?P<0.05?.5.Detection of myocardial cell culture solution NO concentration results:There was no significant difference between the groups?P>0.05?.6.The expression of relevant proteins in the myocardial TGF-?1/Smad2 and PI3K/Akt signaling pathways was determined:The expression of TGF-?1 and Smad2 in 10^-6and10^-7mol/L AngII model group was higher than that in blank group?P<0.05?.Compared with the model group,the TGF-?1 and Smad2 levels in the 10%drug-containing serum group decreased?P<0.05?.The protein expression of P-Akt,Akt and eNOS in 10^-7mol/LAngII model group was lower than that in blank group?P<0.05?.The expression of P-Akt,Akt and eNOS protein in the 10%drug-containing serum group was higher than that in the model group?P<0.05?.The effect of abolishing the 10%drug-containing serum group in the LY294002blocker group was lower than that of the 10%drug-containing serum group?P<0.05?,and the P-Akt,Akt,and eNOS protein expression was decreased?P<0.05?.There was no significant difference in the expression of P-eNOS protein among the groups?P>0.05?.Conclusion:The serum containing Danggui Buxue decoction has a protective effect on Ang?-induced hypertrophic cardiomyocytes,which may be related to the regulation of PI3K/Akt signaling pathway and TGF-?1/Smad2 signaling pathway in cardiomyocytes.