| Objective:Discussion of calcium, magnesium, copper, zinc,vitamin D and soy intervention endemic fluorosis of coal burning on female reproductive function of the ovaries and its possible mechanism of intervention. Methods: 1. Add a free-edible calcium, magnesium, copper, zinc, vitamin D and strengthen woven gold Ward coal soybean peat mix different formulations plus baked corn feed an animal model to build. 2. Dynamic observing and recording the changes of female rats’ teeth; 3. Determination of fluorine content of urine and bone with Ion Selective Electrode Method; 4. Separation of ovarian: ①Weighing, calculate the ovarian weight. ②The change of ovary and granulosa cell apoptosis state was obserced by transmission electron microscope. ③Use immunohistochemical method to observe the expression of apoptosis proteins(Bcl-2、Bax) in ovary. ④The distinct follicl. number and shape on ovary were observed with the HE staining. 5. Executing the rats at different periods(60d, 120 d, 180d), take blood from femoral and separate the serum, Serum levels of E2 were determined by electrochemiluminescence immunoassay; Results: 1. Successfully copied the fluoride group and intervention group in animal models. 2. With the extension of dye fluoride time female dental fluorosis symptoms gradually worsened, the intervention group had mild dental fluorosis, and the occurrence of a variety of elements + soy intervention group dental fluorosis in the intervention group compared with many elements of light. 3. Dyed the same group of fluorine time(60,120,180d): The fluoride group of female urinary fluoride and bone fluoride levels were higher, there was significant(P<0.05), and the fluoride group, two female urinary fluoride intervention group gradually increased, and the latter was significantly higher than the former, were statistically significant(P < 0.05); Compared with the fluoride group, the intervention group alone, combined V intervention group of female bone fluoride decreased, there was statistically significant(P<0.05); The fluoride dose the same category: With time, urine fluoride levels were gradually increased, the fluoride group, the intervention group alone, combined intervention group between fluoride content in urine were statistically significant(P < 0.05). Stained bone fluoride content of fluoride group were statistically increasing(P<0.05); Two bone fluoride content in intervention group were gradually reduced, intervention group alone just 60 d, 120 d bone fluorine content and 180 d was statistically significant(P<0.05), joint intervention group bone fluorine content had statistical significance(P<0.05); 4. Dye fluorine groups of ovarian tissue ultrastructure in granulosa cells and stromal cells apoptosis; Two intervention groups still more apparent in the early stages of a ultrastructure damage, with the extension of intervention time compared with dye fluoride group, the intervention group ultrastructure of follicular granulosa cells and stromal cells of female mice were essentially normal, not seen apoptosis change. Add protein combined intervention of ovarian ultrastructure of lipid drops less than single intervention group obviously. 5. Bax protein expression in Ovarian granulosa cells, dye fluorine 120 d, 180 d, the fluoride group is significantly higher than control group, the intervention group, had statistical significance(P<0.05); Many elements + soy intervention group than in the intervention group a variety of elements, and the 180 d between the two intervention groups was statistically significant difference(P<0.05). The same category with time long, Bax protein expression of ovarian granular cells, dye the fluoride group, both intervention groups gradually increased, in comparing the former group were statistically significant(P<0.05). Individual intervention group 60 d and 180 d was statistically significant(P<0.05). 6. Dye fluoride group with the extension of time, follicular granulosa cells from mild, moderate to severe present progressive change of edema, cell structure and the outline is blurry, atresia of the follicle and corpus luteum degradation increase gradually, mature follicle occasional. Two rare mild edema intervention group of follicular granulosa cells, the cellular structure and cell contour returns to normal, normal ovarian follicles at different levels, atresia of follicles and degradation of corpus luteum was lower than that in group with fluorine; Female atresia follicles number compared with the dye fluoride group, control group 60 d, joint intervention group difference was statistically significant(P < 0.05); 120, 180 d in the control group, the intervention group difference was statistically significant(P<0.05). 7. Dye fluorine time of the same group(60, 120, 180 d) females E2 content is: 60 d, compared with control group, the dye fluoride group gradually increased, the intervention group was statistically significant(P<0.05), the combined intervention group was higher than the control group, dye fluoride group, the individual intervention group was statistically significant(P<0.05). 120 d, 180 d, dye the fluoride group were lower than group, compared with control group, the joint intervention group, with statistical significance(P<0.05), and 180 d single intervention group compared with the joint intervention group was statistically significant(P<0.05); The same category with the extension of time control females E2 content fluctuation change, higher than 120 d, 60 d and 180 d, and 60 d was statistically significant(P<0.05). Dye fluoride group gradually reduce the content of E2, and between groups have statistical significance(P < 0.05). Individual intervention group gradually reduce the content of E2, 60 d, 120 d and 180 d comparison was statistically significant(P<0.05). Joint intervention group gradually reduce the content of E2, 60 d compared to 180 d was statistically significant(P<0.05). Conclusion: 1.Calcium, magnesium, copper, zinc, vitamin D may antagonize fluorosis of coal burning on ovarian reproductive toxicity: Antagonize the toxic effects of excessive fluoride on serum estrogen E2, so close to normal levels of ovarian secretion of E2; Fluorosis can antagonize the growth and development of follicles, so reducing the number of atretic follicles; Antagonized for female ovarian tissue ultrastructure injury, the granulosa cells, stromal cells, mitochondria ultrastructure tends to normal levels; Antagonized fluorosis of coal burning affects proapoptotic protein expression in granulosa cells Bax, reduced the expression of Bax, reduced apoptosis in granulosa. 2. On the basis of calcium, magnesium, copper,zinc, vitamin D can be part of the added protein enhance fluorosis intervention, and showed significantly reduced ovarian ultrastructure of lipiddroplets, apoptotic pro tein Bax expression and decreased the number of atretic follicles, and with prolonged intervention, the better. 3. Many elements + soy intervention and multiple intervention of elements on the female rats ovaries coefficient, primordial follicles, primary follicles, secondary follicles and the number of corpora lutea, ovarian morphology, apoptotic protein Bcl-2 had no significant effect. |
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