The Toxic Effects Of Acrylamide On Mice Testicular Tissue And The Correlation With The Expression Of CK18and Fas, FasL, Caspase-3

ObjectiveThis subject established a mice model of AA subchronic exposure, based on which, reproductive toxicity of AA in the testicular tissue, as well as the expression changes and interrelation with CK18and apoptosis related factors for Fas、FasL and Caspase-3were studied, so as to explore the molecular mechanisms and apoptosis pathways, and provide the experimental basis for germ cell apoptosis pathway and molecular regulation mechanism, as well as the mechanism of the AA reproductive toxicity.MethodsHealth KM mice for5-6weeks randomly divided into4groups, each group for10. Different doses of AA (0,10,20,40mg/kg) peritoneal injection for5weeks, each week for6days, the control group was given equal volume distilled water. The body weight was measured each day. After36days, the mice were killed. Testicular coefficients were calculated, then using histopathological technology (HE dyeing), RT-PCR technology, immunohistochemistry methods respectively detected the expression of CK18and apoptosis related factors for FasL, Caspase-3and Fas on the tissue, mRNA, protein level. We analysis the correlations between CK18and apoptosis related factors finally.Result1. During the acrylamide exposure, the body weight, testicles weight and organs coefficient were decreased, compared with the control, there was statistically significant (p<0.01).2. Compared with the normal control group, HE staining showed that mice testis seminiferous tubes were basic rules, and no significant changes in spermatogenic cells at all levels in the10mg/kg group.20mg/kg group and40mg/kg seminiferous tubes are not in the basic rules, seminiferous epithelium level and all levels of spermatogenic cells depressed, mature sperm within the lumen also depressed in tubes.3. RT-PCR and immunohistochemistry staining showed that CK18mRNA and protein expression of20mg/kg group and40mg/kg group in mouse testis were significantly decreased compared with the normal control group (P<0.01).4. The expression of Fas, FasL, Caspase-3in the testicular tissue scattered a few positive cells in the control group, with10mg/kg group positive cells is not obvious,20mg/kg group and40mg/kg group more positive cells, of which, the IOD value was significantly increased, and statistically significant (P<0.01), compared with the normal control group. Conclusion1. The AA subchronic exposure resulted in the slow weight growth, with testicular coefficient reducing.2. Subchronic exposure to AA can make the pathological changes of the mouse testis structure, reduction of the mature sperm as well as spermatogenic function.3. The expression of CK18in gene and protein level decreased significantly, suggesting the changes of CK18expression may involved in the mechanism of the reproductive toxicity induced by AA.4. The expression of apoptosis related factors for FasL Fas and Caspase-3induced by AA in the mice testicular tissue cell increased, resulting in activating the Fas/FasL apoptosis pathways. Show that the existence of caspase-3-dependent mechanism of apoptosis in AA reproductive toxicity.5. The expression of CK18in the mRNA and protein levels were negatively correlated with the expression of Fas, FasL, Caspase-3, which was expression of the CK18in the mRNA and protein levels downregulated, but the expression of Fas, FasL, Caspase-3upregulated, indicating a correlation between the low expression of CK18and high expression of Fas, FasL, Caspase-3induced by AA in the testicular tissues.