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Acrylamide cytotoxic effects on testes and its relationship with expression of vimentin

Author: ZhangXiaoLing
Tutor: YangJianYi
School: Shanxi Medical
Course: Cell Biology
Keywords: Acrylamide Testis Vimentin Apoptosis
CLC: R114
Type: Master's thesis
Year: 2010
Downloads: 107
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Abstract

Objective To study the experimental intraperitoneal injection, gavage, dermal exposure, acrylamide (AA) on mouse testis cells injury; establish AA infected mouse model to study the AA induced apoptosis in male mice testis and its effects on the expression of vimentin effects, and thus the expression of vimentin germ cell apoptosis; investigate the role of male reproductive toxicity induced AA target cells, reproductive toxicity mechanism of AA to provide laboratory evidence. The first part of the experiment contents and methods will be 48 five-week-old clean healthy male Kunming mice were randomly divided into four groups, namely control group, intraperitoneal exposure group, gavage group and dermal exposure groups 12. Exposure dose of 25 mg / kg, continuous exposure 5d, day 1. Weight change was observed after exposure and testicular factor. In the first six days after the initial exposure line single cell gel electrophoresis experiments, the first 19 days of early testicular germ cell micronucleus test. The second part will be 7 to 8 weeks old 40 clean healthy male Kunming mice were randomly divided into four groups, each 10. With different doses of AA (0,10,20,40 mg / kg) in mice intragastrically 5w, 6 days a week, which 0mg/kg given an equal volume of distilled water, normal control group. Subsequent to initial exposure AA mice were sacrificed after 36 days, using histopathological techniques (HE staining), flow cytometry (FCM), TUNEL method, immunohistochemistry, RT-PCR technique, respectively, from the histological level, cellular level, DNA level, protein and mRNA levels of AA and its relationship with apoptosis induced testicular vimentin expression. The experimental results 1. Peritoneal gavage exposure group and body weight of mice were significantly reduced (P lt; 0.01). Three kinds of mice exposed methods and testicular testicular weight coefficient significantly lower than the control group (P lt; 0.05 or P lt; 0.01). Compared with the control group, each group of mice exposed to single cell gel electrophoresis: testicular cell DNA tail length, tail DNA%, tail moment, Olive tail moment were higher, the differences were statistically significant (P lt; 0.05 or P lt; 0.01). (2) three kinds of mice exposed to ways weight and testicular weight were significantly less than the control group (P lt; 0.05 or P lt; 0.01). And compared with the control group, the difference was not statistically significant testicular coefficient (P gt; 0.05). Gavage group early spermatid micronucleus rate was significantly higher than the control group (P lt; 0.05), while the abdominal skin exposure group exposed groups and early germ cell micronucleus rate was higher than the control group, but the difference was not statistically significant (P gt; 0.05) 3.HE staining showed: 10 mg / kg group and the normal control group, testis seminiferous tubules are arranged basic rules spermatogenic cells did not change significantly. 20 mg / kg and 40 mg / kg dose group, seminiferous tubules irregular, reduced levels of the seminiferous epithelium, spermatogenic cells decreased, decreased sperm maturation within the lumen and vacuolation. 4.FCM results show that compared with the control group, each dose group of cells decreased significantly 1C, 2C cells was significantly increased (P lt; 0.01), and 10 mg / kg and 20 mg / kg dose group 1C and 2C cells, no significant difference in the proportion (P gt; 0.05). 10 mg / kg and 20 mg / kg dose group, the proportion of 4C cells increased compared with the control group, 40 mg / kg group, the proportion of 4C cells lower than the previous two, but still higher, and were statistically significant (P lt; 0.01). Each exposure group 1C: 4C, and 1C: 2C were significantly lower than the control group (P lt; 0.01), but each exposure group pairwise comparisons were not statistically different (P gt; 0.05). 4C: 2C only in the 40 mg / kg dose group compared with the control group decreased significantly different (P lt; 0.01). 5. TUNEL method test results showed that the apoptotic index dose group was significantly higher than the control group (P lt; 0.05). 6 Immunohistochemistry showed, 20mg/kg and 40mg/kg group group Vimentin average optical density was significantly reduced compared with normal control group (P lt; 0.05), 1 Omg / kg decreased compared with normal control group, but not statistically Learn the difference (P gt; 0.05). 7. RT-PCR results showed that compared with normal control group, each dose group vimentin mRNA in mouse testis were significantly lower (P lt; 0.05). Conclusion 1 In the present experimental conditions, the three ways can be caused by exposure AA male mice testicular cells to produce DNA damage, early testicular germ cells in mice an increased incidence of micronuclei and produced by gavage way most sensitive to toxic effects; 2.AA mice can induce testicular seminiferous tubules of spermatogenic cells decreased, decreased sperm maturation within the lumen. 3.AA can reduce the spermatogonial cells into sperm performance, so reducing the number of sperm and sperm cells, so that the process of spermatogenesis disorder. Which induced apoptosis of target cells may be spermatogonia. 4.AA seminiferous epithelium can induce apoptosis index increased in a dose-dependent manner. 5.AA makes the waveform protein and mRNA expression was significantly reduced, and mainly in mesenchymal cells decreased expression of vimentin. Reproductive toxicity speculate AA to AA exposure possible mechanisms that vimentin mRNA expression was decreased, resulting in decreased production of vimentin. 6.AA induced testicular germ cell apoptosis and reduced expression of vimentin has some relevance.

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