| ObjectiveObserve the effect of Decabromodiphenyl ether (BDE-209) exposure on the SerW3 cell morphology, survival rate, cytotoxicity, levels of apoptosis and the endoplasmic reticulum stress related proteins CHOP, GRP78, caspase12 expression. Preliminary discussed the effect of BDE-209 on the SerW3 cell and the mechanism of apoptosis induced by BDE-209.MethodsThe mice Sertoli cell line SerW3 was exposed to the BDE-209, the cell was divided into 5 groups:DMSO, 10μg/ml BDE-209,20μg/ml BDE-209,30μg/ml BDE-209, 40μg/ml BDE-209. After 24h, observe the cell morphology under the inverted microscope, detect the cell survival rate by MTT assay, detect cytotoxicity by LDH method, detect the level of cell apotosis by Annexin V/PI double staining and Hoechst staining and detect the endoplasmic reticulum stress related proteins CHOP, GRP78, caspase12 expression by Western Blot.ResultsThe control group cell growth well, the cell body present columnar or irregular shape, and various ecptomas. Cell nucleus were oval or irregular. The exposure groups cell shape changed, cell membrane vacuolization. Detect the SerW3 cell survival rate by MTT assay after 24h BDE-209 exposure. Compared to the control group, the survival rate of 10μg/ml,20μg/ml,30μg/ml,40μg/ml BDE-209 exposed cell decreased with with statistical significance(P<0.05). In the same dose BDE-209 group, the cell survival rate decreased over time(8h,16h,20h,24h,48h) with statistical significance(P < 0.05). Detect the cytotoxicity by LDH method after 24h BDE-209 exposure. Compared to the control group, the level of LDH of 10μg/ml,20μg/ml,30μg/ml, 40ug/ml BDE-209 exposed cell increased with statistical significance(P<0.05). Detect the level of cell apotosis by Annexin V/PI double staining, compared to the control group, the 20vg/ml BDE-209 group cell apoptosis index increased significantly, and the normal cell ratio decreased. Compared to the 20μg/ml BDE-209 group, the BDE-209 +4-PBA group apoptotic cell ratio decreased and normal cell ratio increased. The Hoechst staining showed that the control group cell nucleus with a normal size and asymmetrical distribution, nucleus nucleoli are visible clearly. With the increase of dosage andages, the apoptotic cell increases, cell membrane vacuolization seriously, the body of the cell was smaller, and more light staining and bright chromatin brought together, the nucleus pycnosis burst. Compared to the control group, ratio of 40μg/ml BDE-209 group cell caspase12/β-actin was increased with statistical difference (P< 0.05), ratio of 30μg/ml and 40 μg/ml BDE-209 group cell CHOP/β-actin and GRP78/β-actin was increased with statistical difference (P<0.05).Conclusion BDE-209 could induce the SerW3 cell apoptosis, change the SerW3 cell morphology and decrease the survival rate, and presented dose-effect relationship. The level of SerW3 cell apoptosis decreased by endoplasmic reticulum inhibitors 4-PBA.ObjectiveIn intro study has proved that BDE-209 could induce the mice Sertoli cell line SerW3 apoptosis. In vivo study to verify BDE-209 could induce mice testis apoptosis by electron microscope and Hoechst staining.MethodsIn vivo,52 four-week-old mice, adaptive feeding a week, were randomly divided into four groups, namely corn oil group, low-dose group(100 mg/kg BW), middle-dose-group(300 mg/kg BW) and high-dose-group(500 mg/kg BW),13 mice in each group. Mice exposed PBDE-209 By gavage, sacrificed after 6 weeks, separated organ on ice, weight and then stored at -80℃. Testicular tissue apoptosis was observed by the electron microscopy and Hoechst staining.ResultsThe mice testis epithelial cells arranged rules, the nuclear envelope and nucleoli were obvious under the electron microscopy. The cell body shape was columnar or irregular with full of cytoplasm, and the endoplasmic reticulum, mitochondria were abundant in the cell, the nuclei were full and located in one side of the cell. In the low dose (100mg/kg/d) group, the cell ultrastructure changed unconspicuously but slight chromatin condensation. In the middle dose group, the number of Sertoli cell decreased, part of the cell mitochondria vacuolization, nuclear chromatin edge accumulation was seen. In high dose(500mg/kg/d) group, the cell changed obviously, the seriously damaged cell chromatin assembly, karyotheca dissolved and nucleus dissolved. The Hoechst staining show that the control group cell with normal size nucleus which located in one side of the cell and unsymmetrical distribution, the nucleoli were obvious. Compared to the control group, the BDE-209 increased the level of cell apoptosis and vacuolation. The body of the cell was smaller and more light staining and bright chromatin brought together, the nucleus pycnosis burst.ConclusionBDE-209 could induce the adolescent male mice testis tissue apoptosis... |
PDF Full Text Request