GDF11 通过下调 CCL11 减轻血管紧张素 II 诱导的肥厚性心肌病和 ANP、BNP 及β-MHC 的表达。
GDF11 Attenuated ANG II-Induced Hypertrophic Cardiomyopathy and Expression of ANP, BNP and Beta-MHC Through Down- Regulating CCL11 in Mice.
机构信息
Department of Cardiology, Gong Li Hospital, 219 Miao Pu Road, Pu Dong New Area, Shanghai 200135, China.
出版信息
Curr Mol Med. 2018;18(10):661-671. doi: 10.2174/1566524019666190204112753.
BACKGROUND
Growth differentiation factor 11 (GDF11) decreases with age, and increased C-C motif chemokine 11 (CCL11) is involved in aging. However, the effects of GDF11 on Angiotensin II (ANG II)-induced hypertrophic cardiomyopathy and expression of markers for volume overload and hypertrophy such as ANP, BNP and beta-MHC, as well as the relationship between GDF11 and CCL11 in hypertrophic cardiomyopathy are unclear. Therefore, the current study aimed to examine the effects of GDF11 on ANG II-induced hypertrophic cardiomyopathy and expression of ANP, BNP and beta-MHC in mice, and explore possible molecular mechanisms.
METHODS
Vectors were constructed and viruses were packaged. Mouse cardiomyocytes were treated with ANG II for 24 h. Meanwhile, mouse cardiomyocytes were divided into 4 groups: (1) control; (2) ANG II; (3) ANG II+GDF11; and (4) ANG II+CCL11. Furthermore, mouse cardiomyocytes were treated with GDF11 and CCL11 proteins for 48 h, respectively. The thickness of IVS and LVPS during systole and diastole were measured by cardiac ultrasound in the mouse model of hypertrophic cardiomyopathy. The relative expression of ANP, BNP, beta-MHC, CCL11 and GDF11 in cardiomyocytes or heart tissue of mice was detected by qPCR or Western blot. 3'- UTR luciferase reporter assay was utilized to examine the relationship between GDF11 and the expression of CCL11.
RESULTS
The expression of ANP, BNP, and beta-MHC in mouse cardiomyocytes was significantly increased after the cells were treated with 800 nM ANG II, which was utilized in the following cell experiments. After ANG II treatment, 0.2 ng/ml GDF11 group displayed the highest inhibition of expression of ANP, BNP and beta-MHC in mouse cardiomyocytes, whereas 50 ng/ml CCL11 group displayed the highest stimulation of the expression. GDF11 at 10 ng/ml significantly decreased the expression of CCL11 in mouse cardiomyocytes as compared to the control group. Mice treated with ANG II had increased thickness of IVS and LVPS during both systole and diastole, which was significantly attenuated by GDF11 overexpression. GDF11 overexpression attenuated the increase in expression of ANP, BNP and beta-MHC in the mice model of hypertrophic cardiomyopathy. The relative serum concentration of GDF11 was markedly decreased, and CCL11 was dramatically increased in mice with hypertrophic cardiomyopathy. GDF11 overexpression restored the serum concentration of GDF11 and CCL11 in the mice model of hypertrophic cardiomyopathy. In addition, GDF11 interference group had markedly increased expression of CCL11, whereas GDF11 overexpression group had significantly decreased expression of CCL11 in luciferase reporter assay.
CONCLUSIONS
GDF11 attenuated ANG II-induced hypertrophic cardiomyopathy and expression of ANP, BNP and beta-MHC through down-regulating CCL11 in mice.
背景
生长分化因子 11（GDF11）随年龄增长而减少，C 型趋化因子配体 11（CCL11）的增加与衰老有关。然而，GDF11 对血管紧张素 II（ANG II）诱导的肥厚性心肌病以及 ANP、BNP 和β-MHC 等容量超负荷和肥大标志物的表达的影响，以及 GDF11 和 CCL11 在肥厚性心肌病中的关系尚不清楚。因此，本研究旨在探讨 GDF11 对 ANG II 诱导的肥厚性心肌病和小鼠 ANP、BNP 和β-MHC 表达的影响，并探讨可能的分子机制。
方法
构建载体并包装病毒。用 ANG II 处理小鼠心肌细胞 24 小时。同时，将小鼠心肌细胞分为 4 组：（1）对照组；（2）ANG II 组；（3）ANG II+GDF11 组；（4）ANG II+CCL11 组。此外，分别用 GDF11 和 CCL11 蛋白处理小鼠心肌细胞 48 小时。通过心脏超声测量肥厚性心肌病小鼠模型中 IVS 和 LVPS 在收缩期和舒张期的厚度。用 qPCR 或 Western blot 检测心肌细胞或小鼠心脏组织中 ANP、BNP、β-MHC、CCL11 和 GDF11 的相对表达。3'-UTR 荧光素酶报告基因检测用于研究 GDF11 与 CCL11 表达的关系。
结果
用 800 nM ANG II 处理细胞后，小鼠心肌细胞中 ANP、BNP 和β-MHC 的表达显著增加，随后进行了细胞实验。ANG II 处理后，0.2ng/ml GDF11 组小鼠心肌细胞中 ANP、BNP 和β-MHC 的表达抑制最明显，而 50ng/ml CCL11 组刺激表达最明显。与对照组相比，10ng/ml 的 GDF11 显著降低了小鼠心肌细胞中 CCL11 的表达。用 ANG II 处理的小鼠 IVS 和 LVPS 在收缩期和舒张期的厚度均增加，而 GDF11 过表达可显著减轻这种情况。GDF11 过表达可减轻肥厚性心肌病小鼠模型中 ANP、BNP 和β-MHC 的表达增加。肥厚性心肌病小鼠血清中 GDF11 的相对浓度明显降低，CCL11 显著增加。GDF11 过表达可恢复肥厚性心肌病小鼠模型中 GDF11 和 CCL11 的血清浓度。此外，在荧光素酶报告基因检测中，GDF11 干扰组 CCL11 的表达明显增加，而 GDF11 过表达组 CCL11 的表达明显减少。
结论
GDF11 通过下调 CCL11 减轻了 ANG II 诱导的肥厚性心肌病和 ANP、BNP 和β-MHC 在小鼠中的表达。
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