Hypoglycemia induced changes in cell death and cell proliferation in the organogenesis stage embryonic mouse heart.
Hypoglycemia induced changes in cell death and cell proliferation in the organogenesis stage embryonic mouse heart.
复制标题
低血糖引起器官发生阶段胚胎小鼠心脏细胞死亡和细胞增殖的变化。
DOI：
10.1002/bdra.20000
复制
发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Smoak,IdaW
中科院分区：
文献类型：
--
作者：
Ghatnekar,GautamS;Barnes,JillA;Dow,JanetL;Smoak,IdaW
糖尿病治疗的副作用之一是低血糖，它会导致心脏发育异常。胚胎心脏细胞对致畸剂诱导的细胞凋亡有很大的抵抗力。方法通过将胚胎日龄（E）9.5（plug = E0.5）的小鼠胚胎在体内或体外暴露于低血糖（30-50 mg/dl葡萄糖）24 h，检测低血糖对胚胎心脏细胞细胞死亡和细胞增殖的影响。在E10.5时通过以下方式评价细胞死亡：1）在切片胚胎中进行两次TUNEL测定以证明DNA片段化; 2）在用Lysotracker染色的整个胚胎中进行共聚焦显微镜检查; 3）在分散的心脏细胞中进行流式细胞术，对TUNEL和肌球蛋白重链（MHC）染色以定量和表征细胞类型易感性;和4）对切片胚胎进行免疫组织化学（MHC）和Western分析，以评估caspase-3活性亚基和p53的潜在参与。通过增殖细胞核抗原（PCNA）的免疫组化和Western分析评价对细胞增殖的影响。E9.5体内低血糖暴露降低了E18.5检查的胚胎的活力。E10.5检查的心脏显示TUNEL和Lysotracker染色增加。在暴露于低血糖的胚胎心脏中，流式细胞术显示TUNEL阳性细胞和TUNEL和MHC双标记细胞增加。caspase-3活性亚基和p53蛋白表达增加，PCNA显着降低，在心脏的胚胎暴露于lowglycempty. CONCLUSIONSSHypoprostimulated胚胎活力，诱导显着的细胞死亡，并减少细胞增殖在E9.5小鼠心脏，这些过程可能涉及活性caspase-3和p53。出生缺陷研究（A部分），2004年。© 2004 Wiley利斯公司
BACKGROUNDHypoglycemia is a side effect of diabetes therapy and causes abnormal heart development. Embryonic heart cells are largely resistant to teratogen‐induced apoptosis.METHODSHypoglycemia was tested for effects on cell death and cell proliferation in embryonic heart cells by exposing mouse embryos on embryonic day (E) 9.5 (plug = E0.5) to hypoglycemia (30–50 mg/dl glucose) in vivo or in vitro for 24 hr. Long‐term effects of in vivo exposure on conceptus viability were evaluated at E18.5. Cell death was evaluated on E10.5 by: 1) two TUNEL assays in sectioned embryos to demonstrate DNA fragmentation; 2) confocal microscopy in whole embryos stained with Lysotracker; 3) flow cytometry in dispersed heart cells stained for TUNEL and myosin heavy chain (MHC) to quantify and characterize cell type susceptibility; and 4) immunohistochemistry (IHC) and Western analysis in sectioned embryos to evaluate potential involvement of caspase‐3 active subunit and p53. Effects on cell proliferation were evaluated by IHC and Western analysis of proliferating cell nuclear antigen (PCNA).RESULTSIn vivo hypoglycemic exposure on E9.5 reduced viability in conceptuses examined on E18.5. Hearts examined on E10.5 demonstrated increased TUNEL and Lysotracker staining. In hearts of embryos exposed to hypoglycemia, flow cytometry demonstrated increased TUNEL‐positive cells and cells dual‐labeled for TUNEL and MHC. Protein expression of caspase‐3 active subunit and p53 was increased and PCNA was markedly reduced in hearts of embryos exposed to hypoglycemia.CONCLUSIONSHypoglycemia reduces embryonic viability, induces significant cell death, and reduces cell proliferation in the E9.5 mouse heart, and these processes may involve active caspase‐3 and p53. Birth Defects Research (Part A), 2004. © 2004 Wiley‐Liss, Inc.