Author + information
- Yi Zhang1,2,
- Chun Li1,
- Qian Zhang3,
- Yan Wu4,
- Qiyan Wang3,
- Hui Meng1,2,
- Jun Li1,
- Yong Wang3 and
- Pengfei Tu1
- 1Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China
- 2School of Chinese materia medica, Beijing University of Chinese Medicine, Beijing, 100102, China
- 3School of Life Sciences, Beijing University of Chinese Medicine, Beijing, China
- 4Institute of Beijing Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China
Baoyuan decoction(BYD), one of the classic traditional Chinese medicine formula, has been validated to have definite protective effect on anti-myocardial apoptosis. However, the mechanisms of its therapeutic effects are yet to be clarified. In this study, we aim to demonstrate that BYD has cardioprotective effect by regulating the expression of Cryab(Crystallin Alpha B) to inhibit myocardial apoptosis both in vivo and in vitro.
Sprague-Dawley (SD) rats were randomly divided into 4 groups: sham group, model group, BYD treatment group and Ginaton(positive drug) treatment group. 7 days after the surgery of left anterior descending (LAD) ligation, heart function was evaluated by 2D echocardiography. The levels of CK-MB and LDH in serum were detected. The expressions of ATF-6, IRE-1, MEKK6, P38 MAPK, MAPKAP-K2, Cryab, Caspase 3, Caspase 9, Bax, Bcl-2 and Cytochrome C were measured by western-blot. The in vitro model of H9C2 cells injured by conditional media of LPS-stimulated macrophages was established to investigate the specific anti-apoptosis mechanisms of BYD. MTT was used to evaluate the cell viability, while Hochest for apoptosis rate. The apoptotic related proteins mentioned above were also detected in different dosage of BYD groups together with positive drug group treated by SB203580, the P38 MAPK inhibitor.
The levels of EF and FS were significantly decreased in model group compared to sham group, suggesting that heart failure model was successfully induced. In BYD treated group, the levels of EF and FS were significantly up-regulated compared with model group. In addition, BYD could decrease cardiac injuring indicators including CK-MB and LDH remarkably compared with model group. Moreover, the expressions of Caspase 3, Caspase 9, Bax and Cytochrome C were down-regulated while the expressions of Bcl-2 and Cryab were up-regulated by the BYD, suggesting that BYD can protect cardiac by apoptosis. The expressions of MEKK6, P38, MAPKAP-K2, ATF6 and IRE-1, the upstream proteins of Cryab, in BYD group were up-regulated to activate Cryab. The results in vitro demonstrated that BYD had significant effect on protecting cell viability and inhibiting the expressions of pro-apoptosis proteins. SB203580 treatment down-regulated the expressions of P38 and Cryab, furthermore, increased the expressions of those pro-apoptosis proteins showed above, suggesting that probably BYD exerts cardioprotective effect through p38/MAPK induced Cryab pathway.
BYD exerts the cardioprotective effect by up-regulating the expression of Cryab to anti-apoptosis either in vivo or in vitro. The underlying mechanism is potentially related to the Cryab pathway. These findings validate a new cardioprotective mechanism and establish foundation for BYD in the clinical application for heart failure.