Objective

2

To explore the mechanism of

Shenfu Yixin Granules

（参附益心颗粒） in the treatment of chronic heart failure （CHF）.

Methods

2

The ionic components of

Shenfu Yixin Granules

were detected by ultra high performance liquid chromatography-quadruped-time-of-flight tandem mass spectrometry. H9c2 cardiomyocytes were divided into normal 1 group， model 1 group，

Shenfu Yixin Granules

low-dose group （SYL），

Shenfu Yixin Granules

high-dose group （SYH）， losartan group， and coenzyme Q10 group. Angiotensin Ⅱ（AngⅡ） was used to induce H9c2 cardiomyocytes injury model in all the groups except for the normal 1 group. Simultaneously， the cells in the SYL and SYH groups were cultured with the medium containing 0.25 mg/ml and 0.5 mg/ml of

Shenfu Yixin Granules

， respectively. The culture medium in the losartan group containd 1×10

-4

mol/L of losartan tablets， and that in the coenzyme Q10 group used 1×10

-4

mol/L of coenzyme Q10 tablets. Each group was cultured for 24 hours. The H9c2 cells were then divided into normal 2 group， model 2 group， transfection model group， transfection

Shenfu Yixin Granules

group （t-SY）， and transfection control group. Except for the normal 2 group and the transfection control group， the other groups used AngⅡto induce H9c2 myocardial cell injury model. Simultaneously， the transfection model group， the t-SY group， and the transfection control group were given lipofectmine 2000 transfection reagent for transfection. The cells in the t-SYgroup was continuously cultured with

Shenfu Yixin Granules

using the dose that had better results before transfection. After that， the cell activity was detected by CCK-8. Mito-tracker fluorescent probe was used to detect mitochondrial content in H9c2 cells. The lactate dehydrogenase （LDH） content， and mRNA expression of cardiomyocyte-related genes including peroxisome proliferator-activated receptor alpha （PPARα）， peroxisome proliferator-activated receptor β/δ （PPARβ/δ）， peroxisome proliferator-activated receptor gamma coactivator 1α （PGC-1α）， nuclear respiratory factor 1 （NRF-1）， nuclear respiratory factor 2 （NRF-2） and estrogen-related receptor （ERR） were detected.

Results

2

There were a total of 387 positive ion substances in

Shenfu Yixin Granules

， and the most abundant ones were Stachydrine， 5-hydroxymethylfurfural and retronecine； 308 qualitative negative ion substances were obtained， and the abundant ones were Danshensu， malic acid， and 4-hydroxybenzoic acid. The dose used in t-SY group was 0.5 mg/ml. Before gene silencing， compared to the normal 1 group， the cell activity and the mRNA expression of each related gene decreased， while the LDH content increased （

P

＜0.05）， and the mitochondrial content decreased in the model group 1. Compared to model 1 group， the cell activity increased and the LDH content decreased in the losartan group， the coenzyme Q10 group， SYH and SYL groups； the mRNA expression of PGC-1α， NRF-1， NRF-2， ERR in the losartan group， that of PPARα， PGC-1α， NRF-1 in the coenzyme Q10 group， that of NRF-1， NRF-2 in the SYL group， and that of PPARα， PGC-1α， NRF-1， NRF-2 and ERR in the SYH group significantly increased （

P

＜0.05）， and the mitochondrial content increased. Compared to SYL group， the cell activity increased while the LDH content decreased in the SYH group， the losartan group and the coenzyme Q10 group （

P

＜0.05）， and the mitochondrial content increased； the mRNA expression of PGC-1αand NRF-2 in the SYH group increased （

P

＜0.05）. After gene silencing， compared to normal 2 group， the cardiomyocyte activity and the mRNA expression of each related gene decreased， while the LDH content increased （

P

＜0.05） in the model 2 group and the transfection model group， and the mitochondrial content decreased. Compared to the transfection model group， there were no significant differences in cardiomyocyte activity， LDH content， and mRNA expression of related genes in the t-SY group （

P

＞0.05）， and the mitochondrial content was not significantly improved. Compared to the SYH group， the cardiomyocyte activity and the mRNA expressions of PPARα， PPARβ/δ， PGC-1α， NRF-1， NRF-2 and ERR were reduced， and the LDH content increased in the t-SY group （

P

＜0.05）.

Conclusion

2

Shenfu Yixin Granules

may improve cardiomyocyte damage in treatment of CHF by regulating PGC-1α signaling pathway related factors. And the active substances mainly include tanshinone， stachydrine， 5-hydroxymethylfurfural， and malic acid.