Lee Seon-Bong, Kim Ju-Hyeong, Cho Myung-Haing, Choe Eun-Sang, Kim Kwang-Sik, Shim Soon-Mi
a Department of Food Science and Technology , Sejong University , Seoul , Republic of Korea.
b Laboratory of Toxicology , College of Veterinary Medicine, Seoul National University , Seoul , Republic of Korea.
J Toxicol Environ Health A. 2017;80(10-12):533-541. doi: 10.1080/15287394.2017.1355863. Epub 2017 Aug 30.
The purpose of the current study was to investigate the effect of two commercial cigarette smoke condensates (CCSC) on oxidative stress and cell cytotoxicity in human brain (T98G) or astrocytes (U-373 MG) in the presence of human brain microvascular endothelial cells (HBMEC). Cell viability of mono-culture of T98G or U-373 MG was markedly decreased in a concentration-dependent manner, and T98G was more susceptible than U-373 MG to CCSC exposure. Cytotoxicity was less prominent when T98G was co-cultured with HBMEC than when T98G was co-cultured with U-373 MG. Significant reduction in trans-epithelial electric resistance (TEER), a biomarker of cellular integrity was noted in HBMEC co-cultured with T98G (HBMEC-T98G co-culture) and U-373 MG co-cultured with T98G (U-373 MG-T98G co-culture) after 24 or 48 hr CCSC exposure, respectively. TEER value of U-373 MG co-cultured with T98G (79-84%) was higher than HBMEC co-cultured with T98G (62-63%) within 120-hr incubation with CCSC. Reactive oxygen species (ROS) generated by CCSC in mono-culture of T98G and U-373 MG reached highest levels at 4 and 16 mg/ml, respectively. ROS production by T98G fell when co-cultured with HBMEC or U-373MG. These findings suggest that adverse consequences of CCSC treatment on brain cells may be protected by blood-brain barrier or astrocytes, but with chronic exposure toxicity may be worsened due to destruction of cellular integrity.
本研究的目的是在人脑血管内皮细胞(HBMEC)存在的情况下,研究两种市售香烟烟雾浓缩物(CCSC)对人脑(T98G)或星形胶质细胞(U - 373 MG)氧化应激和细胞毒性的影响。T98G或U - 373 MG单培养的细胞活力以浓度依赖性方式显著降低,并且T98G比U - 373 MG对CCSC暴露更敏感。当T98G与HBMEC共培养时,细胞毒性比与U - 373 MG共培养时不那么突出。在分别暴露于CCSC 24小时或48小时后,与T98G共培养的HBMEC(HBMEC - T98G共培养)和与T98G共培养的U - 373 MG(U - 373 MG - T98G共培养)中,跨上皮电阻(TEER)显著降低,TEER是细胞完整性的生物标志物。在与CCSC孵育120小时内,与T98G共培养的U - 373 MG的TEER值(79 - 84%)高于与T98G共培养的HBMEC(62 - 63%)。CCSC在T98G和U - 373 MG单培养中产生的活性氧(ROS)分别在4和16 mg/ml时达到最高水平。当与HBMEC或U - 373MG共培养时,T98G产生的ROS减少。这些发现表明,血脑屏障或星形胶质细胞可能保护CCSC处理对脑细胞的不良后果,但长期暴露可能由于细胞完整性的破坏而使毒性恶化。