Li Na, Yang Yue-Jin, Cui He-He, Zhang Qian, Jin Chen, Qian Hai-Yan, Dong Qiu-Ting, Zhang Hao
State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Disease, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
J Cardiovasc Pharmacol. 2014 Mar;63(3):265-73. doi: 10.1097/FJC.0000000000000044.
Tongxinluo (TXL), a traditional Chinese medicine, is widely used to treat cardiovascular diseases in China. Our previous study has demonstrated the pro-survival role of TXL on mesenchymal stem cells (MSCs) in vivo. But whether TXL could decrease apoptosis of MSCs in vitro, and the underlying mechanism are still unknown. Moreover, AMPK/eNOS pathway is crucial in regulating cell apoptosis. Therefore, we designed the study to investigate whether TXL could decrease MSCs apoptosis under hypoxia and serum deprivation (H/SD) conditions and to determine the role of AMPK/eNOS pathway. To test the hypothesis, MSCs were treated with TXL (50-400 μg/mL) under H/SD for 6 hours. For inhibitor studies, the cells were preincubated with AMPK inhibitor compound C. Results indicated that TXL decreased MSCs apoptosis concentration-dependently evidenced by reduced Annexin V+/PI- cells and increased red/green ratio of JC-1. Further, TXL enhanced the phosphorylation of AMPK and eNOS. Whereas, treatment with compound C decreased the phosphorylation of AMPK and eNOS and was accompanied by attenuated anti-apoptotic effect of TXL. In conclusion, TXL protected MSCs against H/SD-induced injury at least in part through the AMPK/eNOS pathway, which provides a novel explanation for the multi-effect of TXL on cardiovascular system.
通心络（TXL）是一种中药，在中国被广泛用于治疗心血管疾病。我们之前的研究已经证明了通心络在体内对间充质干细胞（MSCs）具有促生存作用。但是通心络是否能在体外减少间充质干细胞的凋亡以及潜在机制仍不清楚。此外，AMPK/eNOS通路在调节细胞凋亡中起关键作用。因此，我们设计了本研究，以探讨通心络在缺氧和血清剥夺（H/SD）条件下是否能减少间充质干细胞的凋亡，并确定AMPK/eNOS通路的作用。为了验证这一假设，将间充质干细胞在H/SD条件下用通心络（50 - 400μg/mL）处理6小时。对于抑制剂研究，细胞先用AMPK抑制剂化合物C进行预孵育。结果表明，通心络以浓度依赖性方式减少间充质干细胞凋亡，这通过Annexin V+/PI-细胞减少和JC-1红绿比值增加得以证明。此外，通心络增强了AMPK和eNOS的磷酸化。然而，用化合物C处理降低了AMPK和eNOS的磷酸化，并伴随着通心络抗凋亡作用的减弱。总之，通心络至少部分通过AMPK/eNOS通路保护间充质干细胞免受H/SD诱导的损伤，这为通心络对心血管系统的多重作用提供了新的解释。
