Department of Genetics, Center for Genetics, National Research Institute for Family Planning, Health Department, 12, Dahuisi Road, Haidian, Beijing, 100081, China.
Graduate School, Peking Union Medical College, Beijing, China.
Mol Med. 2021 Feb 10;27(1):13. doi: 10.1186/s10020-021-00267-6.
Gestational diabetes mellitus is a risk factor for congenital heart defects. The article aimed to investigate the expression and roles of MST1, YAP1, Last1/2 and Survivin in modulating HG-induced cardiomyocyte apoptosis and maternal diabetes-induced heart abnormality.
Diabetes mellitus was induced in rats using streptozotocin. The protein expression and phosphorylation analysis in fetal heart tissue was assessed by western blot and immunohistochemical staining. Hoechst 33342 staining assay was performed to explore H9C2 apoptosis. The gene and protein expression in H9C2 cells was assessed by quantitative PCR and western blot. Knockdown of gene expression was assessed by RNA interference.
Our results revealed that increased MST1 protein levels in the heart tissues of the offspring of diabetic rats in vivo and in H9C2 cardiomyocytes under HG treatment in vitro, respectively. Knockdown and overexpression experiments showed that MST1 played a key role in mediating HG-induced apoptosis in cardiomyocytes. Downregulation of YAP1 was associated with HG-induced, MST1-mediated cardiomyocyte apoptosis. Further study showed that MST1 downregulated the protein level of YAP1 through mediation of YAP1 phosphorylation on Ser127 and Ser397; this process also required LATS1/2 participation. MST1 overexpression increased the phosphorylation levels of LATS1/2, which were also shown to be increased in the heart tissues of diabetic offspring. We also found that YAP1 mediated the expression of Survivin during HG-induced apoptosis, and the Survivin-inhibitor YM155 partially inhibited the role of YAP1 in suppressing apoptosis induced by HG in cardiomyocytes.
These findings reveal a regulatory mechanism of MST1/YAP1/Survivin signaling in modulating cardiomyocyte apoptosis in vitro and maternal diabetes-induced congenital heart defects in vivo.
妊娠期糖尿病是先天性心脏缺陷的一个危险因素。本文旨在研究 MST1、YAP1、Last1/2 和 Survivin 在调节高糖诱导的心肌细胞凋亡和母体糖尿病诱导的心脏异常中的表达和作用。
采用链脲佐菌素诱导大鼠糖尿病。通过 Western blot 和免疫组化染色评估胎儿心脏组织中的蛋白表达和磷酸化分析。采用 Hoechst 33342 染色法检测 H9C2 细胞凋亡。通过定量 PCR 和 Western blot 评估 H9C2 细胞中的基因和蛋白表达。采用 RNA 干扰评估基因表达的敲低。
我们的结果表明，糖尿病大鼠子代心脏组织中 MST1 蛋白水平增加，体外高糖处理的 H9C2 心肌细胞中也是如此。敲低和过表达实验表明，MST1 在介导高糖诱导的心肌细胞凋亡中起关键作用。YAP1 的下调与高糖诱导的 MST1 介导的心肌细胞凋亡有关。进一步的研究表明，MST1 通过介导 YAP1 在 Ser127 和 Ser397 上的磷酸化来下调 YAP1 蛋白水平；这个过程还需要 LATS1/2 的参与。MST1 的过表达增加了 LATS1/2 的磷酸化水平，在糖尿病子代的心脏组织中也观察到了这种增加。我们还发现，YAP1 介导 Survivin 在高糖诱导的凋亡中的表达， Survivin 抑制剂 YM155 部分抑制了 YAP1 在抑制心肌细胞中高糖诱导的凋亡中的作用。
这些发现揭示了 MST1/YAP1/Survivin 信号在体外调节心肌细胞凋亡和体内母体糖尿病诱导的先天性心脏缺陷中的调节机制。
