斑马鱼hdac10通过促进自噬提高低温胁迫下的细胞存活

Zebrafish hdac10 Improves Cell Survival Under Low Temperature Stress by Promoting Autophagy

组蛋白去乙酰化酶10(histone deacetylase 10,HDAC10)是一种重要的表观遗传酶,在介导同源重组、调控转录等多种生物过程中发挥作用,然而其在鱼类低温胁迫中的作用尚不清楚。本研究发现低温胁迫下人(Homo sapiens)HEK-293T细胞和斑马鱼(Danio rerio)ZF4细胞中hdac10的表达水平都显著提高。构建了斑马鱼hdac10过表达质粒并转染HEK-293T细胞,Western blot显示外源性斑马鱼hdac10在HEK-293T细胞中成功表达,同时发现hdac10过表达后自噬相关基因BECN1和LC3的mRNA和蛋白水平都显著上调,低温胁迫下过表达外源性hdac10的HEK-293T细胞存活率显著上升(P<0.05),表明低温胁迫下过表达hdac10可以保护HEK-293T细胞。使用表达hdac10-shRNA的慢病毒感染斑马鱼ZF4细胞,RT-qPCR显示ZF4细胞内源性hdac10被显著敲降,hdac10敲降的ZF4细胞中自噬相关基因becn1和lc3的mRNA和蛋白水平都显著下调,低温胁迫下hdac10敲降的ZF4细胞存活率显著下降(P<0.05),表明低温胁迫下hdac10可以保护ZF4细胞。总之,hdac10可以通过上调细胞自噬相关基因的表达来提高细胞在低温胁迫下的存活。本研究为研究鱼类在低温下的抗寒机制提供了科学依据。

HDAC10(histone deacetylase 10)is an important epigenetic enzyme that plays a role in multiple biological processes,such as homologous recombination and regulation of transcription.However,the role of HDAC10 in fish under low temperature stress still remains unclear.In this study,it was showed that the expression of hdac10 in both HEK-293 T cells and ZF4 cells significantly increased under low temperature stress.A zebrafish hdac10 overexpression plasmid was constructed and transfected into HEK-293 T cells.Western blot analysis showed that exogenous hdac10 was successfully expressed in HEK-293 T cells,and the mRNA and protein levels of autophagy related genes,BECN1 and LC3,were significantly upregulated in hdac10 overexpressing HEK-293 T cells,and the survival rate of HEK-293 T cells overexpressing exogenous hdac10 significantly increased under low temperature stress(P<0.05),indicating that zebrafish hdac10 overexpression could protect HEK-293 T cells under low temperature stress.Then ZF4 cells were infected with lentivirus expressing hdac10-shRNA,and RT-qPCR showed that endogenous hdac10 was significantly knocked down in ZF4 cells,the mRNA and protein levels of becn1 and lc3 in ZF4 cells were significantly downregulated by hdac10 knockdown,and the survival rate of ZF4 cells under low temperature stress significantly decreased after hdac10 knockdown(P<0.05),indicating that hdac10 could protect ZF4 cells under low temperature stress.In conclusion,zebrafish hdac10 upregulates the expression of autophagy related genes and improves cell survival under low temperature stress.This study provides a scientific basis for studying the cold resistance mechanism of fish under low temperature.