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Effects on Expression of Adhesion Molecule-related Protein in Uterus of Mice Exposed to Carbon Disulfide during Embryo Implantation

Author: WuYanLing
Tutor: WangZhiPing
School: Shandong University
Course: Epidemiology and Biostatistics,
Keywords: Carbon disulfide Embryotoxicity Embryo implantation Adhesion molecules Hormone
CLC: R714.2
Type: Master's thesis
Year: 2011
Downloads: 14
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Abstract


Background carbon disulfide (Carbon disulfide, CS2) are organic solvents commonly used in industry, is also an important chemical raw materials in contact with a large number of women workers in the viscose fiber production and processing. Epidemiological studies show that: CS2 job workers the rate of spontaneous abortion, early pregnancy loss rate, offspring birth defects rate than the control group was significantly higher, suggesting that CS2 reproductive and developmental toxicity. The preliminary work of our group found that mice exposed in the embryo implantation period (GD4-GD6) in CS2 can significantly reduce the number of embryos implanted, and confirmed that the mice were exposed to CS2 also cause embryonic damage phenomenon occurs. However, CS2 induced embryo toxicity mechanisms are not yet clear. Implantation (embryo implantation), refers to the state of the activated the blastocyst suffer state interaction of the uterus, the trophoblast and endometrial build closer ties, involving the participation of a variety of substances. The embryo research results show that the adhesion molecule integrin v 3 is a marker of endometrial receptivity, and its expression is regulated by progesterone (progesterone, P4) and estradiol (Estradiol, E2); leukemia inhibitory factor ( Leukaemia inhibitory factor, LIF) the endometrium is specifically expressed in the \promote apoptosis of endometrial decidual and involved in the process of embryo implantation. In addition, integrin v 3 upregulate the expression of survivin protein. These results indicate that LIF and survivin integrin v 3 upstream and downstream regulatory proteins. Therefore, this study embryo implantation as an entry point to integrin v 3 and its related proteins for the study of indicators, from the point of view of the endometrial adhesion to explore the mechanism of CS2 embryo toxicity. Research purposes build embryo implantation period at different time points exposed to CS2 induced mouse embryo implantation barriers to animal models, observing detect different mouse embryo development and implantation relationship between the obstacles phenomenon with different exposure time and its time variation; different end point of exposure time pregnant rat uterine tissue integrin beta3, LIF, survivin protein expression levels and serum P4 and E2 concentration of the CS2 exposure caused embryo implantation barriers and adhesion molecules associated protein expression and the relationship between hormone levels and its pathogenic mechanism, in order to reveal the implantation exposed to CS2 embryo toxicity mechanism, provide a theoretical basis for the amendments to the CS2 female workers labor protection measures. Research methods to establish the implantation barriers to a time-dependent animal models female mice by intraperitoneal injection of 10U pregnant mare serum (PMSG) 48 h after injection of 10U of human chorionic gonadotropin (HCG), male and female 1:1 post with cage, the date of inspection the vaginal plug, Yin Shuanyang, remember for pregnant day 1 (GD1). Pregnant rats were randomly divided into GD3, GD4 GD5 GD6 exposed and control groups, in GD3 GD4, GD5 GD6 dose of 631.4mg/kg (0.4LD50) by intraperitoneal injection CS2, the control group received olive oil . Each group was based on the respective exposure time points are located, GD5, GD6, GD7, GD9 the end point to the end of the experiment, weighed and the weight of the embryos, uterus, ovary, liver, spleen, kidney, save uterine tissue samples and serum; The count GD9 end point the number of embryos implanted. 2 detection maternal uterine tissue adhesion molecule expression level and will be exposed group and control group of pregnant mice uterine tissue homogenate supernatant protein quantitative analysis, the use of sodium dodecyl sulfate - polyacrylamide coagulation gel electrophoresis (SDS-PAGE) and Western blot (Western blotting) Determination of adhesion molecule protein relative content. Protein bars with after scanning analysis of the integral absorbance value (integrated Optical Density, IOD). Determination of maternal serum sex hormone levels by enzyme-linked immunosorbent assay (enzyme linked immunosorbent assay, ELISA) determination of the exposed and control groups maternal serum sex hormone levels. 4 statistical analysis using Excel to create the database, using SPSS16.0 software for statistical analysis. Each index homogeneity of variance test: If the homogeneity of variance, one-way ANOVA (ONE-WAY ANOVA) F-test, Dunnett-t test for the experimental group and the control group indicators were compared; heterogeneity of variance, non-parametric statistical methods (Kruskal-Wallis H method). Two independent samples t-test (Independent-samples T-Test) for the experimental group and the control group were compared, the statistical test level was set at alpha = 0.05. Results implantation of exposure CS2 embryo toxic effects due to maternal toxicity: exposure CS2 can lead to the pregnant rats GD9 weight and overall weight weight gain reduced embryo implantation at different time points, but the difference between the net weight gain restructuring does not have statistically significant (P = 0.08). Maternal uterus, ovary, liver, spleen, kidney and other organ weight and organ coefficient was no significant difference between the treatment groups. The relatively small doses caused maternal toxicity effects embryotoxicity: GD4 GD5 exposed to CS2 significantly reduce the number of embryo implantation (P = 0.01 and 0.02); GD4, and GD5 exposed CS2 can be significantly reduced embryo litter weight (P = 0.020 and 0.021), after correction pregnant rats body weight and the number of implanted litter embryonic weight group difference was not statistically significant; implantation exposure CS2 Palace embryo litter weight, litter weight Palace embryo / weight, Palace embryo litter weight / plant into the number had no significant effect. Mouse embryo implantation exposed to CS2 can significantly reduce the number of implantation, the embryo toxicity role. 2.CS2 embryonic implantation at different time points exposed to CS2 exposed to maternal uterine tissue adhesion molecule-associated protein expression can lead to maternal uterine tissue the LIF expression level in the respective first end point was significantly lower (P lt ; 0.05), and GD3 exposure group LIF expression levels in the GD5 and GD6 end point was significantly reduced (P lt; 0.05) Each exposure group in GD7 observe the end of the integrin beta 3 relative content significantly lower than the control group (P lt; 0.05), and the GD3 exposed group GD4 and GD5 observe the end of the integrin beta 3 expression level was significantly decreased (P lt; 0.01) Tip implantation GD3 exposure CS2 integrin beta3 expression of LIF and produce significant interference effect, the integration of the beta3 change appear later in LIF. Different end points of the implantation of CS2 exposure time is not observed significant changes of the uterine tissue expression of survivin. Tip implantation of CS2 exposure to the survivin relatively small embryonic implantation bivariate correlation analysis between the number of maternal uterine tissue adhesion molecule-associated protein expression levels, the implant number of LIF Spearman rank correlation coefficient of 0.327 (P = -0.037), integrin p3 correlation coefficient of 0.289 (P = 0.025), the correlation coefficient with survivin was not statistically significant (r = 0.188, P = 0.151). Prompt the CS2 exposure caused the number of pregnant mice implanted reduce maternal uterine tissue LIF and integrin beta3 expression lower level was positively correlated with no significant relationship between maternal uterine tissue expression of survivin. 3.CS2 exposure to maternal serum sex hormone embryo implantation at different time points (GD3 GD4, GD5) pregnant rats exposed to CS2 the end point GD6 maternal serum P4 level a significant effect (P lt; 0.01 ), GD5 and GD6 exposure CS2 lead to GD9 end point P4 levels decreased (P lt; 0.05). Implantation period of pregnant rats exposed to C82 can lead to GD7 and GD9 observed the endpoint serum E2 levels were significantly lower (P lt; 0.05), GD3 and GD5, exposed end point of GD6 E2 levels produce different degrees of interference (P = 0.016 and 0.055 ). Prompt CS2 exposure the maternal serum P4 level is in GD6 GD9 E2 mainly in GD7 GD9. Implantation between the number of maternal serum P4 and E2 levels bivariate correlation analysis showed that the number of implanted with P4 and E2 Spearman rank correlation coefficients were 0.119 and 0.102 (P 0.365 and 0.438), indicating that exposure to CS2 The pregnant mice embryos implanted decrease in the number of no significant association between maternal hormone levels. Conclusion mouse embryo implantation exposed to CS2 can lead to significant embryotoxicity, GD4 GD5 exposure caused embryo loss significantly. (2) implantation exposed to CS2 LIF and integrin p3 expression levels can be reduced in mouse uterine tissue and implanted in the decrease in the number of significant positive correlation between the degree of adhesion molecules of low expression. Prompt decrease in protein levels may be an important mechanism of to CS2 exposure to embryo toxicity 3 implantation exposure CS2 can reduce the serum P4 and E2 levels, hormone levels change significantly change in adhesion molecules the LIF and integration prime beta3 after, and the implant was no significant correlation between the degree of reduction in the number and hormone levels. Prompt implantation mouse embryos exposed to CS2 caused by LIF and integrin adhesion molecule expression leads to low embryo implantation disorders, hormone levels change its smaller effect. This phenomenon needs further study confirmed.

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CLC: > Medicine, health > Obstetrics and Gynaecology > Obstetrics > Pathological pregnancy ( abnormal pregnancy )
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