Liu H, Yu C, Yu H, Zhong L, Wang Y, Liu J, Zhang S, Sun J, Duan L, Gong L, Yang J
Yuhuangding Hospital, No. 20 of Yuhuangding East Road, Yantai City, Shandong Province, China.
Second People's Hospital of Zhangdian District, Zibo, Shandong Province, China.
Herz. 2018 Jun;43(4):346-351. doi: 10.1007/s00059-017-4564-3. Epub 2017 Apr 25.
Cerebral dopamine neurotrophic factor (CDNF) has been studied in animal models of Parkinson's disease, where it was shown to repair and protect dopamine neurons. Alongside its neurotrophic activity, it can also localize in the endoplasmic reticulum (ER) acting as an ER stress response (ERSR) protein to maintain ER homeostasis. Since ER stress plays a major role in the development and progression of cardiovascular diseases, we investigated the role of CDNF in cardiomyocytes during ER stress.
Initially, the expression of CDNF was tested by treating H9c2 cells with various concentrations of tunicamycin (TM) and performing reverse-transcriptase polymerase chain reaction and Western blotting. To evaluate the overexpression of CDNF in cardiomyocytes, H9c2 cells were transfected with pcDNA-CDNF and analyzed by Western blotting and indirect immunofluorescence microscopy. The effects of CDNF on cardiomyocytes during ER stress were analyzed with CCK-8 method and TUNEL staining using cells transfected with pcDNA-CDNF or pcDNA3.1. The percentage of TUNEL-positive cells was quantified as the apoptotic level.
Our results showed that CDNF protein expression can be induced by activation of ER stress in cultured cardiomyocytes. Moreover, overexpression of CDNF improved cell viability and protected cardiomyocytes from apoptosis induced by ER stress.
The findings presented here contribute toward identifying the physiological functions of CDNF in cardiovascular diseases.
脑源性多巴胺神经营养因子(CDNF)已在帕金森病动物模型中得到研究,研究表明它能修复和保护多巴胺能神经元。除了其神经营养活性外,它还可定位于内质网(ER),作为一种内质网应激反应(ERSR)蛋白来维持内质网稳态。由于内质网应激在心血管疾病的发生和发展中起主要作用,我们研究了内质网应激期间CDNF在心肌细胞中的作用。
首先,通过用不同浓度的衣霉素(TM)处理H9c2细胞并进行逆转录聚合酶链反应和蛋白质印迹法来检测CDNF的表达。为了评估CDNF在心肌细胞中的过表达情况,用pcDNA-CDNF转染H9c2细胞,并通过蛋白质印迹法和间接免疫荧光显微镜进行分析。使用用pcDNA-CDNF或pcDNA3.1转染的细胞,通过CCK-8法和TUNEL染色分析内质网应激期间CDNF对心肌细胞的影响。将TUNEL阳性细胞的百分比定量为凋亡水平。
我们的结果表明,内质网应激激活可诱导培养的心肌细胞中CDNF蛋白表达。此外,CDNF的过表达提高了细胞活力,并保护心肌细胞免受内质网应激诱导的凋亡。
本文的研究结果有助于确定CDNF在心血管疾病中的生理功能。