Effect Of Danggui Buxue Decoction Medicated Serum On The Apoptosis Of Angiotensin â…¡-induced Cardiomyocyte Hypertrophy

Objective: To explore the effect of Danggui Buxue Decoction on chronic heart failureby observing the effect of Danggui Buxue Decoction medicated serum on the apoptosis ofhypertrophic cardiomyocytes induced by angiotensinⅡ.Methods: The models of the hypertrophic cardiomyocytes induced by angiotensinⅡwereconfirmed by observing the content of the total cellular protein. There were four groups in thisexperiment: control group, AngⅡgroup (10-7mol/L), AngⅡ+valsartan group (10-6mol/L) andAngⅡ+medicated serum group (8.4g/kg). MTT method was used to observe the effect ofeach treatment measures on cardiomyocytes activity at0h,12h,24h,48h,72h and96hrespectively, and determine optimum time of activties. At this time point screening theoptimal concentration of medicated serum of Danggui Buxue Decoction. Hoechst33342staining was used to detect cardiomyocytes apoptosis rate. Cardiomyocytes DNA damage wasdetected by Single-cell gel electrophoresis. Mitochondrial injuries of the myocardium wereobserved with the electron microscope. Western Blotting was used to detect the change of Bax,Bcl-2and p53proteins.Results:1. Determination of protein content: Compared with the control group, the totalprotein of AngⅡ group increased greatly, which has significant difference (P<0.05).2. The different time point of cardiomyocytes activity: In0h, the cell activity among eachgroup was no statistical significance (P>0.05). In12h and24h, the cell activity of groupsincreased, and the level in AngⅡgroup was higher than the others (P<0.05). In48h,72h and96h, the cell activity of all groups decreased. Meanwhile the cell activity in AngⅡ group waslower than those in control group and AngⅡ+valsartan group after72h (P<0.05). At the sixtime point, there was no statistics significance among control group and AngⅡ+valsartangroup (P<0.05).3. The best concentration of Danggui Buxue Decoction containing serum: In48h, the cellactivity of medicated serum40%group and medicated serum20%group were much lowerthan that in control group and AngⅡgroup (P<0.05), the cell activity of medicated serum10%group was much lower than that of AngⅡgroup (P<0.05), and at the same time thecontrol group changed little (P<0.05), the cell activity of medicated serum5%group was much higher than that of control group(P<0.05), and at the same time the AngⅡ groupchanged little(P<0.05).4. The cardiomyocytes apoptosis rate: Compared with the control group, cardiomyocytesapoptosis rate in AngⅡgroup significantly increased (P<0.05). In AngⅡ+valsartan group andAngⅡ+medicated serum group, cardiomyocytes apoptosis rate was significantly lower(P<0.05).5. Single cell gel electrophoresis test results: In AngⅡgroup, the cardiomyocytes valuesof H-DNA%significantly lower, and the values of T-DNA%, TL and TM were significantlyhigher than that of the control group(P<0.05). In Ang II+medicated serum group andAngⅡ+valsartan group, the cardiomyocytes values of H-DNA%significantly higher, and thevalues of T-DNA%, TL and TM were significantly lower than that of the AngⅡ group(P<0.05).6. The electron microscope results: The cardiomyocytes mitochondria’s membrane wascomplete and clear in the control group. The membrane showed significant vacuolization andblurred or fractured or deleted crest fusion in AngⅡgroup. In AngⅡ+valsartan group andAngⅡ+medicated serum group, cardiomyocytes mitochondrial ultrastructure showed mildedema, and at the same time the control group changed little.7. Western Blotting results: Compared with the control group, p53and Bax proteinsexpression in AngⅡgroup significantly increased (P<0.05), but Bcl-2protein expressionsignificantly decreased (P<0.05), the ratio of Bcl-2/Bax significantly decreased (P<0.05). InAng Ⅱ+valsartan group and Ang Ⅱ+medicated serum group, p53and Bax Proteinsexpression was significantly lower (P<0.05), but Bcl-2proteins expression was significantlyhigher (P<0.05), and the ratio of Bcl-2/Bax was significantly higher (P<0.05).Conclusions: Danggui Buxue Decoction medicated serum could inhibit the apoptosis ofAng II-induced cardiomyocytes, the mechanism may be related to reduce DNA damage,protect of mitochondrial and regulate Bcl-2, Bax and p53protein expression of hypertrophycardiomyocytes.