[1]梁关凤,李素娟,邱晓霞,等.激活Sonic hedgehog通路改善缺血缺氧心肌细胞DNA损伤[J].中国药理学通报,2018,(09):1235-1242.[doi:10.3969/j.issn.1001-1978.2018.09.011]
 LIANG Guan-feng,LI Su-juan,QIU Xiao-xia,et al.Activation of Sonic hedgehog signaling in ventricular cardiomyocytes exerts DNA protection against hypoxia[J].Chinese Pharmacological Bulletin,2018,(09):1235-1242.[doi:10.3969/j.issn.1001-1978.2018.09.011]
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激活Sonic hedgehog通路改善缺血缺氧心肌细胞DNA损伤()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2018年09期
页码:
1235-1242
栏目:
论著
出版日期:
2018-08-26

文章信息/Info

Title:
Activation of Sonic hedgehog signaling in ventricular cardiomyocytes exerts DNA protection against hypoxia
文章编号:
1001-1978(2018)09-1235-08
作者:
梁关凤1李素娟2邱晓霞1张贵平1罗健东1袁文常3侯 宁1
1.广州医科大学药学院药理学教研室,广东 广州 511436; 2.广东省第二人民医院药学部,广东 广州 510000; 3.广州医科大学附属第五医院检验科,广东 广州 510700
Author(s):
LIANG Guan-feng1 LI Su-juan2 QIU Xiao-xia1 ZHANG Gui-ping1 LUO Jian-dong1 YUAN Wen-chang3 HOU Ning1
1.Dept of Pharmacology, Guangzhou Medical University, Guangzhou 511436, China; 2.Dept of Pharmacy, Guangdong Second Provincial General Hospital, Guangzhou 510000, China; 3.Dept of Clinical Laboratory,the Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou 510700,China
关键词:
原代乳鼠心肌细胞 Sonic hedgehog通路 缺氧 DNA损伤 细胞凋亡 GANT61 SAG1.3
Keywords:
primary neonatal rat cardiomyocytes Sonic hedgehog pathway hypoxia DNA damage cell apoptosis GANT61 SAG1.3
分类号:
R-332; R322.11; R329.25; R342.3; R542.22; R845.22; R977.6
DOI:
10.3969/j.issn.1001-1978.2018.09.011
文献标志码:
A
摘要:
目的 探究Sonic hedgehog通路在糖氧剥夺(oxygen and glucose deprivation, OGD)诱导的乳鼠心肌细胞DNA损伤和细胞凋亡中的作用及机制。方法 原代培养乳鼠心肌细胞,建立OGD的缺血缺氧模型,分别给予Sonic hedgehog通路激动剂SAG1.3(0.002 μmol·L-1 )、抑制剂GANT61(0.005 μmol·L-1 )处理6、12 h,MTT法检测细胞生存率; Western blot 检测DNA损伤标志物γH2AX、DNA损伤反应蛋白ATM、ATR,以及细胞凋亡相关蛋白p53、凋亡标志物cleaved-caspase-3、Bcl-2、Bax表达; 免疫荧光检测γH2AX表达。结果 与正常组相比,OGD组心肌细胞γH2AX、p-ATM表达呈时间依赖性增加, OGD 6 h达到高峰,OGD 12 h后表达逐渐下降; p-p53、cleaved-caspase-3表达增加,Bcl-2/Bax比例下降; 与OGD 6 h、OGD 12 h组相比,给予SAG1.3组OGD心肌细胞γH2AX、p-ATM表达明显下降,心肌细胞存活率上升,p-p53、cleaved-caspase-3表达下降, Bcl-2/Bax比例上升; GANT61组OGD心肌细胞较OGD组γH2AX、p-ATM表达明显上升,细胞存活率下降,p-p53、cleaved-caspase-3表达上升, Bcl-2/Bax比例下降。结论 激活Sonic hedgehog通路可以通过抑制ATM磷酸化,减少γH2AX表达,改善缺血缺氧诱导的心肌细胞DNA损伤,抑制细胞凋亡。
Abstract:
Aim To investigate the protective roles of sonic hedgehog(Shh)signaling pathway in hypoxia-induced DNA damage with the neonatal rat cardiomyocytes. Methods The hypoxia model on neonatal cardiomyocytes was established with one to two days old Sprague Dawley rats by deprivation of oxygen and glucose(OGD). After pretreated with Shh pathway agonist SAG1.3 or antagonist GANT61, the survival rates of cardiomyocytes were assayed by MTT after OGD 6 hours or 12 hours. The protein levels of Shh pathway, phosphorylated histone H2AX at serine 139(γH2AX), phosphorylated ATM(p-ATM), phosphorylated p53(p-p53), cleaved-caspase-3, Bcl-2 and Bax were detected by Western blot. The γH2AX foci was detected by immunofluorescence. Results Compared to control group, the protein expression of γH2AX, p-ATM, cleaved-caspase-3, p-p53 in OGD cardiomyocytes significantly increased, and Bcl-2/Bax ratio proportionally decreased. Particularly, the expression of γH2AX, p-ATM was highest at OGD 6 h, and then gradually declined after OGD 12 h. After SAG1.3 pretreatment, the expression of γH2AX, p-ATM, cleaved-caspase-3 and p-p53 dramatically decreased and the Bcl2/Bax ratio increased in OGD 6 h or OGD 12 h cardiomyocytes. On the contrary, in GANT61 pretreatment group, the expression of γH2AX, p-ATM, cleaved-caspase-3 and p-p53 significantly increased and the Bcl-2/Bax ratio decreased compared to the OGD 6 h or OGD 12 h cardiomyocytes. Conclusion The activation of Shh pathway protects cardiomyocytes against hypoxia-induced apoptosis through inhibition of DNA damage.

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备注/Memo

备注/Memo:
收稿日期:2018-04-20,修回日期:2018-06-10
基金项目:国家自然科学基金资助项目(No 81573433,81773720); 广东省高等学校优秀青年教师培养计划(No YQ2015135); 广东省普通高校特色创新项目(Q17024053); 广州市教育科研计划项目(No 1201610064)
作者简介:梁关凤(1991-),女,硕士生,研究方向:心血管药理学,E-mail: 763957221@qq.com;
侯 宁(1981-),女,博士,教授,硕士生导师,研究方向:心血管药理学,通讯作者,E-mail:houning-2003@163.com
更新日期/Last Update: 2018-08-26