Abstract
Thirdhand smoke (THS) is a new term for the toxins in cigarette smoke that linger in the environment long after the cigarettes are extinguished. The effects of THS exposure on male reproduction have not yet been studied. In this study, metabolic changes in male germ cell lines (GC-2 and TM-4) were analyzed after THS treatment for 24 h. THS-loaded chromatography paper samples were generated in a laboratory chamber system and extracted in DMEM. At a paper: DMEM ratio of 50 μg/ml, cell viability in both cell lines was normal, as measured by the MTT assay and markers of cytotoxicity, cell cycle, apoptosis and ROS production were normal as measured by quantitative immunofluorescence. Metabolomic analysis was performed on methanol extracts of GC-2 and TM-4 cells. Glutathione metabolism in GC-2 cells, and nucleic acid and ammonia metabolism in TM-4 cells, was changed significantly by THS treatment. RT-PCR analyses of mRNA for enzyme genes Gss and Ggt in GC-2 cells, and TK, SMS and Glna in TM-4 cells reinforced these findings, showing changes in the levels of enzymes involved in the relevant pathways. In conclusion, exposure to THS at very low concentrations caused distinct metabolic changes in two different types of male reproductive cell lines.
Highlights
A population-based study from South Korea showed an increase of prevalence of respiratory symptoms in children exposed to THS due to parental smoking[8]
In the present study, we investigated the effects of THS exposure on metabolism in two male reproductive cell lines, GC-2 and TM-4
The GC-2 cell line was originally derived from immortalized murine spermatogonia, and the TM-4 cell line was derived from murine sertoli cells
Summary
A population-based study from South Korea showed an increase of prevalence of respiratory symptoms in children exposed to THS due to parental smoking[8] These data have provided preliminary evidence suggesting that THS exposure can induce health effects. Sertoli cells provide both structural and nutritional support to germ cells, and maintain the spermatogenous microenvironment during spermatogenesis. Both cell lines are useful models in testing the effects of chemicals on male reproductive system, including the study of metabolomics[9,10,11]. Results showed that low concentrations of THS in which no significant effects of THS on general cytotoxicity and multi-parameter cytotoxicity could alter metabolism in GC-2 and TM-4 cells, indicating potential effects of THS on male reproduction
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