Effects of Alcohol on Apoptosis in Spermatogenic Cells and the Expression of Survivin and Caspase-3 in It
|School||Tianjin Medical University|
|Keywords||Alcohol Spermatogenic cells Apoptosis Survivin Caspase-3 Immunohistochemistry|
Objective: To study alcohol feeding on mice growth and development, testicular and spermatogenic function to understand the technology of the preparation of mouse testis paraffin sections, HE staining. Master situ nick end labeling (TUNEL) detection spermatogenic cell apoptosis and immunohistochemical staining techniques detected specimens to explore each experimental group Caspase-3 protein and Survivin protein expression and alcohol caused spermatogenic correlation of apoptosis. Methods: The experiment with distilled water, red wine, white wine of low-dose and high-dose mice were given orally press 0.01ml / g. The body weight of mice were measured weekly during the experiment and observe the state changes caused by the mice in each group growth and development and alcohol. 3 spermatogenic cycle successfully established the mouse alcohol toxicity model. The mice were decapitated and immediate measurement of bilateral testicular weight. Preparation of paraffin sections of mouse testis using HE staining to observe the changes in each group of mice testis seminiferous tubules and spermatogenic cell morphology and TUNEL assay groups spermatogenic cell apoptosis index (AI) changes. Then detected by immunohistochemical SP method Caspase-3 protein and Survivin protein expression in each experimental group, using a semi-quantitative analysis of immunoreactivity score (immunoreactivity score, IRS) IRS: 0: negative (-), 1 ~ 4: weakly positive (), from 5 to 8 points: moderately positive () ,9-12: strongly positive (). SPSS17.0 statistical software for statistical processing of the experimental data, which is a measure of data analysis of variance, ranked data nonparametric Kruskal-Wallis test, the relationship between the two variables: Spearman rank correlation analysis, each experimental group small. mouse body weight, testicular weight and spermatogenic cell apoptosis index, caspase-3 protein and Survivin protein expression statistical analysis. Results: In this study, about three spermatogenic cycle 0.01mL / g alcohol gavage successfully established model of alcohol toxicity in mice. Experimental control group and wines mice during normal appetite, mental state, alcohol toxicity mice drunken phenomenon. Red wine gavage group average weight gain in mice up alcohol toxicity of high-dose group increased at least, from the mice in each group the average weight gain in terms of the changes, including high-dose alcohol toxicity in mice weight gain was significantly less than the low-dose group mice ((P = 0.003), with increasing liquor dose, mice weight gain less obvious. simultaneous detection of the relative weight of each group of mice testes (bilateral testes relative to their body weight before the experiment mice) and HE staining under the seminiferous tubules diameter changes, but also presents the basic consistency of results. using TUNEL assay mice in each group spermatogenic cell apoptosis index, distilled water the intragastric group spermatogenic cell apoptosis index was 0.1394 ± 0.0104 high in the red wine group (0.1016 ± 0.0124, P = -0.000), but lower than alcohol toxicity group (.2213 ± 0.0149,0.2582 ± 0.0177, respectively, P = 0.000, P = 0.000), in which the average red wine group spermatogenic cells highest lowest apoptotic index, high doses of liquor group, and along with the average increase in spermatogenic cell apoptosis index of liquor dose is gradually increasing. immunohistochemistry results showed that alcohol toxicity group mouse testis Caspase-3 protein expression compared with the control group and the red wine group, and with increasing alcohol dose and its expression, the stronger; Survivn protein expression on the contrary Wine mice testis Caspase-3 protein expression is lower than that of other groups, even lower than the normal control group positive expression of Survivin protein is most significant with the increase in the degree of apoptosis of spermatogenic cells of mice in each group, the stronger expression of caspase-3 protein, showed a positive correlation (r = 0.717, P lt; 0.01); while Survivn the weaker protein, showed a negative correlation (r = -0.631, P lt; 0.01) Conclusion: The amount of red wine gavage mammalian growth and development, testicular spermatogenic function promote, inhibit alcohol toxicity role, and with the increase in liquor dose, inhibition of the more obvious changes in caspase-3 and Survivn protein expression and alcohol caused spermatogenic cell apoptosis is closely related to, but its mechanism of action may be associated with some of the ingredients in the wine , but still are not sure, need to experiment further confirmed that this experiment may be able to change people's drinking, as well as for the clinical treatment of male infertility to provide new ideas.