Involvement of mitochondria-mediated apoptosis in ethylbenzene-induced renal toxicity in rat.
Ethylbenzene is an important industrial chemical that has recently been classified as a possible human carcinogen (International Agency of Research on Cancer class 2B), but the available data do not support the genotoxic mechanism of ethylbenzene-induced tumors in kidney. We investigated the effects of ethylbenzene on renal ultrastructure and explored the nongenotoxic mechanism of mitochondria-mediated apoptosis pathway. Forty male Sprague-Dawley rats were used as a vivo model with ethylbenzene inhalation for 13 weeks, and the metabolites of ethylbenzene, mandelic acid (MA), and phenylglyoxylic acid (PGA) in urine were examined by high-performance liquid chromatography. Meanwhile, the ultrastructure of renal tubular epithelial cells was observed, and cell apoptosis was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. Furthermore, we investigated the expression levels of messenger RNA (mRNA) and protein of bax, bcl-2, cytochrome c, caspase-9, and caspase-3 in rat kidney. With respect to levels of MA, PGA, and MA + PGA, a significant dose-dependent increase was observed in 4335 and 6500 mg/m(3) ethylbenzene-treated groups against the control group. The mitochondria of renal tubular epithelial cells became a compact and vacuolar structure in 6500 mg/m(3) ethylbenzene-treated group, and ethylbenzene induced a significant increase in the number of apoptotic cells as compared to the control group. In addition, enhanced mRNA and protein expression levels of all measured genes were observed in various ethylbenzene-treated groups except the decreased bcl-2 expression levels. Our results indicated that ethylbenzene may induce apoptosis of renal tubular epithelial cells via mitochondria-mediated apoptotic pathways. MA and PGA in urine might be a parameter of biological dose in vivo after ethylbenzene inhalation.
Tianjin Centers for Disease Control and Prevention, Tianjin, People's Republic of China.
Ming
M
Zhang
Yanrang
Y
Wang
Qian
Q
Wang
Junyu
J
Yang
Deyi
D
Yang
Jing
J
Liu
Jianguo
J
Li
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001555:Benzene Derivatives / Q000493:pharmacokinetics / Q000633:toxicity*; D015415:Biomarkers / Q000378:metabolism; D001835:Body Weight / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D015870:Gene Expression / Q000187:drug effects; D006038:Glyoxylates / Q000652:urine; D020287:In Situ Nick-End Labeling; D019570:Inhalation Exposure; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007674:Kidney Diseases / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D007684:Kidney Tubules / Q000187:drug effects / Q000473:pathology; D008297:Male; D008333:Mandelic Acids / Q000652:urine; D008928:Mitochondria / Q000187:drug effects* / Q000648:ultrastructure; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D001555:Benzene Derivatives; D015415:Biomarkers; D006038:Glyoxylates; D008333:Mandelic Acids; D011506:Proteins; D012333:RNA, Messenger; C012482:phenylglyoxylic acid; C004912:ethylbenzene; C037938:mandelic acid