DJ-1蛋白调控乳鼠心肌细胞内质网应激反应的研究

Modulation of Endoplasmic Reticulum Stress Mediated by DJ-1 in Neonatal Cardiomyocytes

【目的】探讨DJ-1蛋白对心肌细胞内质网应激的调控作用。【方法】在原代培养心肌细胞中采用苯肾上腺素(PE)、毒胡萝卜素(TG)诱导心肌细胞内质网应激。通过腺病毒转染过表达DJ-1蛋白,应用荧光定量PCR及Western blotting分别检测在PE及TG作用下,心肌细胞内质网应激标记物GRP78和calreticulin的m RNA及蛋白表达改变,明确DJ-1对心肌细胞内质网应激的作用。【结果】PE、TG均可引起GRP78及calreticulin的m RNA及蛋白表达升高,提示二者均可导致心肌细胞的内质网应激反应。应用腺病毒转染在心肌细胞中过表达DJ-1蛋白,而过表达DJ-1显著下调内质网应激标记物GRP78及calreticulin的m RNA及蛋白表达,提示DJ-1可抑制PE及TG导致的心肌细胞的内质网应激反应。【结论】DJ-1可抑制心肌细胞中PE及TG诱导的内质网应激。

【Objective】 To study how DJ-1 modulates endoplasmic reticulum stress（ERS） in neonatal cardiomyocytes. 【Method】In neonatal cardiomyocytes, thapsigargin（TG） and phenylephrine（PE） were used to induce ERS. Overexpression of DJ-1 protein was mediated by adenovirus transfection. The m RNA and protein levels of ERS markers including GRP78 and calreticulinin cardiomyocytes under PE or TG stress were detected using Q-PCR and Western blotting. 【Results】 TG and PE significantly induced the ERS markers including the m RNA and protein levels of GRP78 and calreticulin, indicating that TG and PE are the inducer of cardiac ERS. The overexpression of DJ-1 was achieved by the utilization of adenovirus. We discovered that overexpression of DJ-1 markedly reduced the m RNA and protein levels of GRP78 and calreticulin, indicating that DJ-1 is a negative modulator of ERS induced by TG and PE.【Conclusion】 DJ-1 inhibits ERS induced by TG and PE in neonatal cardiomyocytes.