Department of Cardiology, Second Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Department of Cardiology, Rongcheng People's Hospital, Rongcheng, Shandong, China.
J Cell Biochem. 2020 Nov;121(11):4337-4346. doi: 10.1002/jcb.29630. Epub 2020 Jan 31.
Diabetic cardiomyopathy can cause cardiac dysfunction and eventually lead to heart failure and sudden death. Long noncoding RNA (lncRNA) Gas5 has been reported to play a function in cardiomyocyte. Here we studied the function of Gas5 on newborn mouse cardiomyocyte (NMC) apoptosis to detect its molecular mechanism. High-glucose treatment was implemented to induce the apoptosis of NMC in this study. And terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, JC-1 assay, and flow cytometry analysis were conducted to know about the apoptosis of NMC when Gas5 and Tcf3 were silenced. Meanwhile, RNA pull-down assay and luciferase reporter assay were conducted to verify the binding of RNAs. Finally, rescue assay was implemented to evaluate the influence on apoptosis situation affected by competing endogenous RNA pathways. Tcf3 was found to bind to the Gas5 promoter to activate the expression of Gas5. Meanwhile, Gas5 and Tcf3 were both found to promote the apoptosis of NMC. Also, mmu-miR-320-3p could bind to Gas5 and Tcf3. Moreover, the Gas5/miR-320-3p/Tcf3 pathway was found to modulate the apoptosis of NMC. In conclusion, Tcf3-activated lncRNA Gas5 regulates NMC apoptosis in diabetic cardiomyopathy.
糖尿病性心肌病可导致心脏功能障碍，最终导致心力衰竭和猝死。长链非编码 RNA (lncRNA) Gas5 已被报道在心肌细胞中发挥作用。在这里，我们研究了 Gas5 对新生小鼠心肌细胞 (NMC) 凋亡的作用，以检测其分子机制。本研究采用高糖处理诱导 NMC 凋亡。并进行末端脱氧核苷酸转移酶 dUTP 缺口末端标记测定、JC-1 测定和流式细胞术分析，以了解 Gas5 和 Tcf3 沉默时 NMC 的凋亡情况。同时，进行 RNA 下拉测定和荧光素酶报告测定以验证 RNA 的结合。最后，进行挽救测定以评估竞争内源性 RNA 通路对凋亡情况的影响。发现 Tcf3 与 Gas5 启动子结合以激活 Gas5 的表达。同时，发现 Gas5 和 Tcf3 均促进 NMC 的凋亡。此外，mmu-miR-320-3p 可以与 Gas5 和 Tcf3 结合。此外，还发现 Gas5/miR-320-3p/Tcf3 通路调节 NMC 的凋亡。总之，Tcf3 激活的 lncRNA Gas5 调节糖尿病性心肌病中的 NMC 凋亡。
