文章摘要
去甲氧基姜黄素对毒胡萝卜素诱导乳鼠心肌细胞内质网应激的影响
Influence of Demethoxycurcumin on Endoplasmic Reticulum Stress in Neonatal Rat Myocardial Cells Induced by Thapsigargin
投稿时间:2020-06-13  录用日期:2020-06-18
DOI:
中文关键词: 去甲氧基姜黄素  毒胡萝卜素内酯  心肌细胞  内质网应激
英文关键词: Demethoxycurcumin  Thapsigargin  Myocardial Cells  endoplasmic reticulum stress
基金项目:黑龙江省自然科学基金面上项目:H2017059,内质网应激-自噬信号通路介导DMC保护心肌缺血再灌注损伤的机制研究。
作者单位E-mail
崔健昆 黑龙江中医药大学 904224307@qq.com 
耿乃志 黑龙江中医药大学  
孟凡吉 哈尔滨医科大学  
施丽春 黑龙江中医药大学  
田耕 黑龙江中医药大学  
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中文摘要:
      目的 探讨去甲氧基姜黄素(Demethoxycurcumin,DMC)对毒胡萝卜素内酯(Thapsigargin,TG))诱导的乳鼠心肌细胞内质网应激( Endoplasmic Reticulum Stress,ERS) 的作用和机制,及其对相关分子葡萄糖调节蛋白78( glucose regulated protein of 78kD,GRP78) 和C/EBP环磷酸腺苷反应元件结合转录因子同源蛋白(CAAT/enhancer binding protein homologous protein,CHOP)的影响。方法 体外培养乳鼠心肌细胞,随机分为4 组: ①对照组细胞不做任何药物处理;②模型组加入1μmol/L TG处理24h,建立ERS模型;③DMC低浓度组同时加入1μmol/L TG和10μmol/L DMC处理24h; ④DMC高浓度组同时加入1μmol/L TG和30μmol/L DMC处理24h。采用CCK8法检测心肌细胞活性,流氏细胞法检测心肌细胞凋亡,检测细胞匀浆中乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)的含量,Western Blot法检测GRP78和CHOP蛋白表达。结果 与对照组相比,模型组细胞活性显著降低,细胞凋亡率明显增加,LDH及MDA 含量升高,而SOD活力则降低,GRP78和CHOP蛋白表达显著升高,P<0.05;与模型组比较,不同浓度DMC干预后,细胞活性升高,细胞凋亡率明显减少,LDH及MDA含量显著降低,SOD活力则显著升高,并且GRP78和CHOP蛋白表达降低,P<0.05。结论 去甲氧基姜黄素通过抑制GRP78和CHOP蛋白表达来减轻TG所致的内质网应激损伤,减少细胞凋亡,从而对心肌细胞起到保护作用。
英文摘要:
      Objective To explore the effect and mechanism of Demethoxycurcumin (DMC) on Endoplasmic Reticulum Stress (ERS) induced by Thapsigargin (TG) in neonatal rat cardiomyocytes, and its influence on related molecules glucose regulated protein of 78kD (GRP78) and C/EBP cyclic adenosine phosphate response element binding transcription factor homologous protein (CAAT/enhancer binding protein homologous protein, CHOP). Methods Cultured neonatal rat cardiomyocytes were randomly divided into 4 groups:①Control group cells were not treated with any drugs; ②Model group added 1μmol/L TG treatment for 24h to establish ERS model; ③DMC low concentration group added 1μmol/L TG and 10μmol/L DMC treatment for 24h at the same time; ④DMC high concentration group added 1μmol /L TG and 30μmol/L DMC simultaneously for 24h.The CCK8 method was used to detect the activity of cardiomyocytes, and the flow cytometry was used to detect cardiomyocyte apoptosis. The content of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD) in cell homogenate were tested. Western Blot Method was used to detect the expression of GRP78 and CHOP protein. Results Compared with the control group, the activity of the cells in the model group significantly decreased, the apoptosis rate significantly increased, the content of LDH and MDA increased, and the activity of SOD decreased, and the expression of GRP78 and CHOP protein significantly increased, P<0.05; compared with the model group, after intervention with different concentrations of DMC,the cell activity increased, the apoptosis rate decreased significantly, the content of LDH and MDA decreased significantly, the activity of SOD increased significantly, and the expression of GRP78 and CHOP protein decreased, P<0.05. Conclusion Demethoxycurcumin can alleviate the injury of TG-induced Endoplasmic Reticulum Stress and reduce apoptosis by inhibiting the expression of GRP78 and CHOP proteins, thereby protecting cardiomyocytes.
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