title: Effects of Cigarette Smoke Condensate on Oxidative Stress, Apoptotic Cell Death, and HIV Replication in Human Monocytic Cells.
abstract: While cigarette smoking is prevalent amongst HIV-infected patients, the effects of cigarette smoke constituents in cells of myeloid lineage are poorly known. Recently, we have shown that nicotine induces oxidative stress through cytochrome P450 (CYP) 2A6-mediated pathway in U937 monocytic cells. The present study was designed to examine the effect of cigarette smoke condensate (CSC), which contains majority of tobacco constituents, on oxidative stress, cytotoxicity, expression of CYP1A1, and/or HIV-1 replication in HIV-infected (U1) and uninfected U937 cells. The effects of CSC on induction of CYP1 enzymes in HIV-infected primary macrophages were also analyzed. The results showed that the CSC-mediated increase in production of reactive oxygen species (ROS) in U937 cells is dose- and time-dependent. Moreover, CSC treatment was found to induce cytotoxicity in U937 cells through the apoptotic pathway via activation of caspase-3. Importantly, pretreatment with vitamin C blocked the CSC-mediated production of ROS and induction of caspase-3 activity. In U1 cells, acute treatment of CSC increased ROS production at 6H (>2-fold) and both ROS (>2 fold) and HIV-1 replication (>3-fold) after chronic treatment. The CSC mediated effects were associated with robust induction in the expression of CYP1A1 mRNA upon acute CSC treatment of U937 and U1 cells (>20-fold), and upon chronic CSC treatment to U1 cells (>30-fold). In addition, the CYP1A1 induction in U937 cells was mediated through the aromatic hydrocarbon receptor pathway. Lastly, CSC, which is known to increase viral replication in primary macrophages, was also found to induce CYP1 enzymes in HIV-infected primary macrophages. While mRNA levels of both CYP1A1 and CYP1B1 were elevated following CSC treatment, only CYP1B1 protein levels were increased in HIV-infected primary macrophages. In conclusion, these results suggest a possible association between oxidative stress, CYP1 expression, and viral replication in CSC-treated cells of myeloid lineage. This study warrants a closer examination of the role of CYP1B1 in smoking-mediated enhanced HIV replication.
authors.affiliation: Department of pharmaceutical sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America.
authors.forename: Pss
authors.initials: P
authors.lastname: Rao
authors.affiliation: Division of pharmacology and toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri, United States of America.
authors.forename: Anusha
authors.initials: A
authors.lastname: Ande
authors.affiliation: Department of pharmaceutical sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America.
authors.forename: Namita
authors.initials: N
authors.lastname: Sinha
authors.affiliation: Division of pharmacology and toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri, United States of America.
authors.forename: Anil
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Department of pharmaceutical sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America.
authors.forename: Santosh
authors.initials: S
authors.lastname: Kumar
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology*; D006801:Humans; D008264:Macrophages / Q000378:metabolism; D009000:Monocytes / Q000821:virology*; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012907:Smoking / Q000009:adverse effects*; D020298:U937 Cells; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D017382:Reactive Oxygen Species; D019363:Cytochrome P-450 CYP1A1; D053148:Caspase 3