title: The effect of tobacco smoke exposure on the generation of reactive oxygen species and cellular membrane damage using co-culture model of blood brain barrier with astrocytes.
abstract: Brain tissue is known to be vulnerable to the exposure by tobacco smoke. Tobacco smoke can induce generation of reactive oxygen species (ROS), causing inflammatory activity and blood-brain barrier (BBB) impairment. The aim of the present study was to investigate the effect of tobacco smoke on cell cytotoxicity, generation of ROS, and cellular membrane damage in astrocytes and BBB using a co-culture system. Cell viability of U373MG cells was reduced in a dose-dependent manner, ranging from 96.7% to 40.3% by tobacco smoke condensate (TSC). Cell viability of U373MG co-cultured with human brain microvascular endothelial cells (HBMECs) was 104.9% at the IC50value of TSC. Trans-epithelial electric resistance values drastically decreased 80% following 12-h incubation. The value was maintained until 48 h and then increased at 72-h incubation (85%). It then decreased to 75% at 120 h. Generation of ROS increased in a dose-dependent manner, ranging from 102.7% to 107.9%, when various concentrations of TSC (4-16 mg/mL) were administered to the U373MG monoculture. When TSC was added into U373MG co-cultured with HBMECs, production of ROS ranged from 101.7% to 102.6%, slightly increasing over 12 h. Maximum exposure-generated ROS of 104.8% was reached at 24 h. Cell cytotoxicity and oxidative stress levels in the U373MG co-culture model system with HBMECs were lower than U373MG monoculture. HBMECs effectively acted as a barrier to protect the astrocytes (U373MG) from toxicity of TSC.
authors.affiliation: 1 Department of Food Science and Technology, Sejong University, Gwangjin-gu, Seoul, Republic of Korea.
authors.forename: Seung-Beom
authors.initials: SB
authors.lastname: Seo
authors.affiliation: 2 Department of Biological Sciences, Pusan National University, Gumjeong-gu, Pusan, Republic of Korea.
authors.forename: Eun Sang
authors.initials: ES
authors.lastname: Choe
authors.affiliation: 3 Pediatric Infectious Diseases, Johns Hopkins University, Baltimore, MD, USA.
authors.forename: Kwang-Sik
authors.initials: KS
authors.lastname: Kim
authors.affiliation: 1 Department of Food Science and Technology, Sejong University, Gwangjin-gu, Seoul, Republic of Korea.
authors.forename: Soon-Mi
authors.initials: SM
authors.lastname: Shim
mesh_terms: D001253:Astrocytes / Q000187:drug effects*; D001812:Blood-Brain Barrier / Q000187:drug effects*; D001921:Brain / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000187:drug effects; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012906:Smoke / Q000009:adverse effects*
chemical_list: D017382:Reactive Oxygen Species; D012906:Smoke
keywords: Tobacco smoke condensate; astrocyte; blood–brain barrier; cellular integrity; reactive oxygen species