浙江农业学报 ›› 2019, Vol. 31 ›› Issue (9): 1446-1452.DOI: 10.3969/j.issn.1004-1524.2019.09.07

• 动物科学 • 上一篇    下一篇

DMSO与DMF对H9C2心肌细胞毒性及STAT3信号通路的影响

杨敏, 张凯, 李建喜, 王磊, 张康, 仇正英, 王学智*   

  1. 中国农业科学院 兰州畜牧与兽药研究所,甘肃省中兽药工程技术研究中心,甘肃 兰州 730050
  • 收稿日期:2019-02-14 出版日期:2019-09-25 发布日期:2019-10-11
  • 通讯作者: *王学智,E-mail:wangxz628@sina.com
  • 作者简介:杨敏(1993—),女,四川罗江人,硕士研究生,主要从事动物临床中兽医学研究。E-mail: yangmin1074335657@163.com
  • 基金资助:
    国家自然科学基金(31702288); 中央级公益性科研院所基本科研业务费专项资金(1610322016013)

Effects of DMSO and DMF on myocardial cytotoxicity and STAT3 signaling pathway in H9C2

YANG Min, ZHANG Kai, LI Jianxi, WANG Lei, ZHANG Kang, QIU Zhengying, WANG Xuezhi*   

  1. Lanzhou Institute of Animal Husbandry and Veterinary Drugs, Chinese Academy of Agricultural Sciences, Engineering & Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province, Lanzhou 730050, China
  • Received:2019-02-14 Online:2019-09-25 Published:2019-10-11

摘要: 为了筛选有机溶剂DMSO与DMF对H9C2心肌细胞毒性作用的最低浓度,探讨DMSO与DMF对心肌细胞信号转导与转录因子3(signal transducer and activator of transcription 3,STAT3)信号通路的影响,通过四氮唑盐比色分析法(MTT法)试验,检测DMSO和DMF在体积分数为0.1%、0.2%、0.3%、0.5%、0.7%、0.9%、1%、2%、3%时作用24、48、72 h心肌细胞的存活率变化;Western blot法检测STAT3和Bcl-2蛋白,对比DMSO和DMF分别在0%、0.2%、0.3%、0.5%体积分数下STAT3蛋白表达量的差异性。结果表明,同一DMSO和DMF体积分数条件下,随着时间增加,细胞存活率下降,在同一时间随着DMSO和DMF体积分数的增加,心肌细胞存活率减小,Bcl-2和STAT3蛋白含量减少,DMSO体积分数为0.3%和0.5%时,细胞存活率有统计学意义(P<0.05),STAT3和Bcl-2蛋白水平比对照组低(P<0.01),但DMSO体积分数为0.2%,细胞存活率与STAT3、Bcl-2表达对比对照组,均无统计学意义(P>0.05);当DMF体积分数为0.3%和0.5%时,细胞存活率有统计学意义(P<0.05),STAT3和Bcl-2蛋白水平比对照组低(P<0.01),当DMF体积分数为0.2%时,细胞存活率无统计学意义(P>0.05),STAT3蛋白表达对比对照组,具有统计学意义(P<0.01)。与对照组(不含DMSO和DMF)比较,DMSO和DMF对H9C2心肌细胞具有毒性作用,随浓度增加,细胞存活率降低,Bcl-2蛋白含量降低,可能与STAT3蛋白下调相关,MTT法检测结果与Western blot试验结果不完全一致,MTT法具有不稳定性。

关键词: H9C2心肌细胞, DMSO, DMF, MTT, STAT3

Abstract: In order to screen the lowest concentration of toxic effects of organic solvents DMSO and DMF on H9C2 cardiomyocytes, and to investigate the effects of DMSO and DMF on signal transduction and transcription factor 3 (signal transducer and activator of transcription 3, STAT3) signal pathway, the survival rates of cardiomyocytes exposed to DMSO and DMF at volume fraction of 0.1%, 0.2%, 0.3%, 0.5%, 0.7%, 0.9%, 1%, 2%, 3% for 24 h, 48 h and 72 h, respectively, were detected by the tetrazolium salt colorimetric assay (MTT method). And the survival rate of STAT3 and Bcl-2 protein was determined by Western blot method, and the expression of STAT3 protein was compared between DMSO and DMF at 0%, 0.2%, 0.3%, 0.5% volume fraction, respectively. At the same time, the survival rate of cardiomyocytes decreased with the increase of the volume percentage of DMSO and DMF, and the content of Bcl-2 and STAT3 protein decreased. When the volume fraction of DMSO was 0.3% and 0.5%, the cell survival rate was significantly lower than that of the control group (P<0.05). The protein levels of STAT3 and Bcl-2 were lower than those of the control group (P<0.05). The survival rate of cardiomyocytes decreased with the increase of the volume percentage of DMSO and DMF, and the content of Bcl-2 and STAT3 protein decreased at the same time. However, when the volume fraction of DMSO was 0.2%, there was no significant difference in the cell survival rate and the expression of STAT3 and Bcl-2 compared with the control group (P>0.05). When the volume fraction of DMF was 0.3% and 0.5%, the cell survival rate was statistically significant (P<0.05), and the protein levels of STAT3 and Bcl-2 were lower than those in the control group. When the volume fraction of DMF was 0.2%, the cell survival rate was not statistically significant (P>0.05). The expression of STAT3 protein was significantly higher than that in the control group (P<0.01). Compared with the control group (without DMSO and DMF), DMSO and DMF had toxic effects on H9C2 cardiomyocytes. With the increase of concentration, the cell survival rate decreased and the content of Bcl-2 protein decreased, which may be related to the down-regulation of STAT3 protein. The results of MTT assay were not completely consistent with those of Western blot test, and MTT assay was unstable.

Key words: H9C2 cardiomyocytes, DMSO, DMF, MTT, STAT3

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