College of Veterinary Medicine, Nanjing Agricultural University, 1 Weigang, Nanjing, 210095, China.
Mol Cell Biochem. 2018 Dec;449(1-2):195-206. doi: 10.1007/s11010-018-3356-2. Epub 2018 Apr 18.
In this report, we investigate the protective mechanism of co-enzyme Q10 on chicken primary myocardial cells during heat stress. Morphological observations indicate that addition of co-enzyme Q10 protects myocardial cells from heat stress, reduces the damage of mitochondria and nucleus, and decreases the mean number of vacuolated mitochondria. We have previously shown that co-enzyme Q10 can protect myocardial cells by upregulating the expression of Hsp70. Therefore, signaling pathways involved in this process were explored. No changes of total MAPK protein (P38MAPK, JNK, ERK) expression in the experimental groups were detected, with the exception of total JNK1. Co-enzyme Q10 failed to increase the expression of JNK1 compared to the HS group which was treated with heat stress only. Addition of Q10 upregulated the expression of p-P38MAPK, p-JNK, and p-ERK1. Inhibitors of P38MAPK and JNK, SB203580 and SP600125, respectively, weakened the upregulation of Hsp70 by co-enzyme Q10, indicating that MAPK pathways participate in the Hsp70 upregulation by co-enzyme Q10. Co-enzyme Q10 upregulates the expression of p-MEK3/6 and p-MEK4, but not p-MEK7 during heat stress. Expression of p-PKCα and p-PKCβ1 was also elevated following the addition of co-enzyme Q10 during heat stress, and addition of PKC inhibitors decreased the expression of Hsp70 induced by co-enzyme Q10. This confirms that PKC is also associated with the upregulation of Hsp70. In HS+Q10 group, addition of SP600125 or SB203580 could increase cell apoptosis under heat stress. Our results suggest that co-enzyme Q10 upregulates the expression of Hsp70 during heat stress to protect chicken primary myocardial cells via the PKC-MEK3/4/6-P38MAPK/JNK pathways.
在本报告中,我们研究了辅酶 Q10 在热应激条件下对鸡原代心肌细胞的保护机制。形态学观察表明,添加辅酶 Q10 可保护心肌细胞免受热应激损伤,减少线粒体和细胞核的损伤,并降低空泡化线粒体的平均数量。我们之前已经表明,辅酶 Q10 通过上调 Hsp70 的表达来保护心肌细胞。因此,我们探索了参与这一过程的信号通路。实验组中总 MAPK 蛋白(P38MAPK、JNK、ERK)的表达没有变化,除了总 JNK1。与仅接受热应激处理的 HS 组相比,辅酶 Q10 未能增加 JNK1 的表达。添加 Q10 可上调 p-P38MAPK、p-JNK 和 p-ERK1 的表达。P38MAPK 和 JNK 的抑制剂 SB203580 和 SP600125 分别削弱了辅酶 Q10 对 Hsp70 的上调作用,表明 MAPK 通路参与了辅酶 Q10 对 Hsp70 的上调作用。辅酶 Q10 在热应激条件下可上调 p-MEK3/6 和 p-MEK4 的表达,但不能上调 p-MEK7。添加辅酶 Q10 后,p-PKCα 和 p-PKCβ1 的表达也升高,添加 PKC 抑制剂可降低辅酶 Q10 诱导的 Hsp70 表达。这证实了 PKC 也与 Hsp70 的上调有关。在 HS+Q10 组中,添加 SP600125 或 SB203580 可在热应激下增加细胞凋亡。我们的结果表明,辅酶 Q10 在热应激条件下通过 PKC-MEK3/4/6-P38MAPK/JNK 通路上调 Hsp70 的表达,从而保护鸡原代心肌细胞。