Sumanasekera Wasana K, Tran David M, Sumanasekera Thimira U, Le Nolan, Dao Halle T, Rokosh Gregg D
Department of Pharmaceutical Sciences, Sullivan University College of Pharmacy, 2100 Gardiner lane, Louisville, KY, 40205, USA,
Cell Biol Toxicol. 2014 Apr;30(2):113-25. doi: 10.1007/s10565-014-9273-6. Epub 2014 Mar 16.
Cigarette smoking is a major risk factor for numerous diseases including cardiovascular diseases. Exposure to cigarette smoke (CS) leads to increased cardiovascular risk, myocardial injury, and mortality. Stem cell therapy is one of the promising therapeutic options available to treat myocardial injuries. Understanding the impact of cigarette smoke extract (CSE) on stem cell function would be valuable in determining the risk passed on during transplant. In this study, the impact of CSE on cardiac stem cell (CSC) functions was investigated using c-kit+ rat cardiac stem cells as the experimental model. Here, we hypothesized that CSE attenuates CSC membrane integrity, causes cytotoxicity, and affects many CSC functions via multiple mechanisms including modulation of extracellular stress-regulated kinase (ERK) (44/42) signaling and oxidative stress. The effects of CSE on CSCs were examined in vitro. Based on a published method, CSE was prepared. CSE-induced ERK signaling was detected by western blotting. CSE-induced modulation of catalase activity was also measured. Functional modulations due to CSE were examined via several methods including Apostain, BrdU, and LDH assays. In agreement with the CSE-induced activation of ERK, CSE-induced reduction in viability, migration, and increase in both cytotoxicity and para-cellular permeability were observed in CSCs. These results suggest that CSE impaired CSC responses that contribute to decreased ability of CSC to respond to stress or injury leading to exacerbation of the damage. Our findings will contribute to the understanding of the discipline and might contribute to the development of stem cell therapy approaches in the future.
吸烟是包括心血管疾病在内的众多疾病的主要危险因素。接触香烟烟雾（CS）会导致心血管风险增加、心肌损伤和死亡率上升。干细胞疗法是治疗心肌损伤的一种有前景的治疗选择。了解香烟烟雾提取物（CSE）对干细胞功能的影响对于确定移植过程中传递的风险具有重要意义。在本研究中，以c-kit+大鼠心脏干细胞为实验模型，研究了CSE对心脏干细胞（CSC）功能的影响。在此，我们假设CSE会削弱CSC膜完整性，导致细胞毒性，并通过多种机制影响许多CSC功能，包括调节细胞外应激调节激酶（ERK）（44/42）信号通路和氧化应激。在体外检测了CSE对CSCs的影响。根据已发表的方法制备CSE。通过蛋白质印迹法检测CSE诱导的ERK信号。还测量了CSE诱导的过氧化氢酶活性调节。通过包括Apostain、BrdU和LDH测定在内的几种方法检查了CSE引起的功能调节。与CSE诱导的ERK激活一致，在CSCs中观察到CSE诱导的活力降低、迁移减少以及细胞毒性和细胞旁通透性增加。这些结果表明，CSE损害了CSC反应，导致CSC应对应激或损伤的能力下降，从而加剧损伤。我们的研究结果将有助于该学科的理解，并可能有助于未来干细胞治疗方法的发展。
