Tang Shu, Wen Qiang, Liu Peiqing, Zhu Zhenfeng, Li Na, Zhang Xiaojian, Kan Quancheng
Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University Zhengzhou 450052, Henan, China.
Department of Clinical Pharmacology, Basic Medical College of Zhengzhou University Zhengzhou 450002, Henan, China.
Int J Clin Exp Med. 2015 Aug 15;8(8):12617-23. eCollection 2015.
This study aims to explore the effects of the traditional Chinese medicine monomer cryptotanshinone (CTS) on the expression levels of inflammatory factors in myocardial cells caused by Ang II and its mechanism.
The neonatal rat myocardial cells were cultured in vitro in this study. Their purities were identified by immunocytochemical method. The cellular viability in different groups was determined by MTT assay. The levels of TNF-α and IL-6 in the supernatant of cell culture were detected with ELISA method. The levels of intracellular reactive oxygen species (ROS) were detected by Dihydrogen ethidium (DHE) staining method. The location changes of NF-κB in cells were detected by immunofluorescence method.
The purity of primary cultured neonatal rat myocardial cells was over 95%, CTS had no obvious effect on the viability of cells while it inhibited the increased levels of TNF-α, IL-6 and ROS caused by Ang II with dose dependent. NF-κB mainly distributed in the cytoplasmic region in normal cells, it translocated to the nucleus after Ang II stimulation while CTS inhibited the translocation.
CTS could inhibit the inflammatory factors such as TNF-α and IL-6 in myocardial cells induced by Ang II with dose dependent, its mechanism may be related with that CTS could decrease the levels of ROS in myocardial cells and inhibit NF-κB translocation into the nucleus.
本研究旨在探讨中药单体隐丹参酮(CTS)对血管紧张素II(Ang II)所致心肌细胞炎症因子表达水平的影响及其机制。
本研究体外培养新生大鼠心肌细胞,采用免疫细胞化学方法鉴定其纯度,采用MTT法检测不同组细胞活力,采用ELISA法检测细胞培养上清液中TNF-α和IL-6水平,采用二氢乙锭(DHE)染色法检测细胞内活性氧(ROS)水平,采用免疫荧光法检测细胞中NF-κB的定位变化。
原代培养的新生大鼠心肌细胞纯度超过95%,CTS对细胞活力无明显影响,但能剂量依赖性抑制Ang II所致的TNF-α、IL-6和ROS水平升高。正常细胞中NF-κB主要分布于细胞质区域,Ang II刺激后其转位至细胞核,而CTS抑制了这种转位。
CTS能剂量依赖性抑制Ang II诱导的心肌细胞中TNF-α和IL-6等炎症因子,其机制可能与CTS降低心肌细胞中ROS水平及抑制NF-κB转位入核有关。