营养元素和蛋白联合干预氟中毒对雌鼠卵巢功能作用
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摘要
目的探讨钙、镁、铜、锌、维生素D和蛋白联合干预燃煤型氟中毒对雌鼠卵巢功能变化。方法 120只体重80 g左右的清洁级SD雌性大鼠为研究对象,随机分为对照组、染氟组、多种元素组、多种元素+蛋白组,每组30只。各模型组大鼠自由食用不同配方的病区原煤加拌泥煤烘烤的玉米饲料。分别于染氟60,120,180 d处死雌鼠,分离双侧卵巢。免疫组化法检测卵巢颗粒细胞凋亡蛋白Bcl-2和Bax的表达。HE染色光镜下观察卵巢组织形态、各级卵泡数量变化。结果卵巢颗粒细胞Bcl-2蛋白表达量差异无显著性。各组与染氟组Bax蛋白比较,120 d和180 d差异有显著性(P<0.05),180 d两干预组比较差异有显著性(P<0.05)。各组内比较,染氟组Bax蛋白表达量逐渐增多,差异有显著性(P<0.05)。多种元素组60 d与180 d比较差异有显著性(P<0.05)。染氟组随时间延长,卵巢颗粒细胞呈现渐进性水肿的变化,闭锁的卵泡和黄体退化逐渐增多。两干预组卵巢颗粒细胞少见轻度水肿,正常的各级卵泡多见,闭锁的卵泡和退化的黄体明显降低;与染氟组相比,60 d对照组和多种元素+蛋白组闭锁卵泡降低,差异有显著性(P<0.05);120 d和180 d对照组、两干预组差异有显著性(P<0.05)。结论钙、镁、铜、锌、维生素D可拮抗燃煤型氟中毒对卵巢生殖毒性降低Bax蛋白的表达量,降低闭锁卵泡数量,同时添加蛋白联合干预效果更佳。
Objective To study effects of Ca,Mg,Cu,Zn,VDcombined with protein on ovarian functions in female rats with coal-burning fluorosis. Methods A total of 120 clean-grade SD female rats with body weights of about 80 g were used. Rats were randomly divided into control,fluorosis,element,and element + protein groups with 30 rats ineach group. Model groups were fed edible raw coal mixed with peat and different formulas of zhijin roasted corn feed. Rats were sacrificed at 60,120,and 180 days,and bilateral ovaries were harvested. Expression of apoptosis proteins Bcl-2 and Bax in ovarian granulosa cells was observed by immunohistochemistry. HE staining was used to observe ovarian tissue morphology and changes in follicle and corpus luteum numbers. Results There was no statistical significance in the protein expression of Bcl-2 in ovarian granulosa cells. The expression of Bax in each group was compared with that in the fluorosis group,and a significant difference was found at 120 and 180 days( P < 0. 05). There were significant differences at 180 days between the two intervention groups( P < 0. 05). Compared within each group,Bax expression in the fluorosis group gradually increased and the difference was significant( P < 0. 05). There was a significant difference at 60 and 180 days for various elements( P < 0. 05). Over a prolonged time in the fluoride group,ovarian granulosa cells showed progressive edema,atretic follicles,and corpus luteum degradation,and mature follicles were decreased significantly.Ovarian granular cells in the two intervention groups showed rare mild edema and normal numbers of follicles at all levels.Atresia of follicles and degeneration of the corpus luteum were decreased significantly. Compared with the fluorosis group,there were significant differences in atresia follicles between control and multiple element + protein groups at 60 days,and the control group and two intervention groups at 120 and 180 days( P < 0. 05). Conclusions Ca,Mg,Cu,Zn,and VD antagonize the reproductive toxicity of coal-burning fluorosis and reduce protein expression of Bax and the number of atresia follicles. And the effects of intervention combined with protein can be better.
Objective To study effects of Ca,Mg,Cu,Zn,VDcombined with protein on ovarian functions in female rats with coal-burning fluorosis. Methods A total of 120 clean-grade SD female rats with body weights of about 80 g were used. Rats were randomly divided into control,fluorosis,element,and element + protein groups with 30 rats ineach group. Model groups were fed edible raw coal mixed with peat and different formulas of zhijin roasted corn feed. Rats were sacrificed at 60,120,and 180 days,and bilateral ovaries were harvested. Expression of apoptosis proteins Bcl-2 and Bax in ovarian granulosa cells was observed by immunohistochemistry. HE staining was used to observe ovarian tissue morphology and changes in follicle and corpus luteum numbers. Results There was no statistical significance in the protein expression of Bcl-2 in ovarian granulosa cells. The expression of Bax in each group was compared with that in the fluorosis group,and a significant difference was found at 120 and 180 days( P < 0. 05). There were significant differences at 180 days between the two intervention groups( P < 0. 05). Compared within each group,Bax expression in the fluorosis group gradually increased and the difference was significant( P < 0. 05). There was a significant difference at 60 and 180 days for various elements( P < 0. 05). Over a prolonged time in the fluoride group,ovarian granulosa cells showed progressive edema,atretic follicles,and corpus luteum degradation,and mature follicles were decreased significantly.Ovarian granular cells in the two intervention groups showed rare mild edema and normal numbers of follicles at all levels.Atresia of follicles and degeneration of the corpus luteum were decreased significantly. Compared with the fluorosis group,there were significant differences in atresia follicles between control and multiple element + protein groups at 60 days,and the control group and two intervention groups at 120 and 180 days( P < 0. 05). Conclusions Ca,Mg,Cu,Zn,and VD antagonize the reproductive toxicity of coal-burning fluorosis and reduce protein expression of Bax and the number of atresia follicles. And the effects of intervention combined with protein can be better.
引文
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[11]Celino FT,Yamaguchi S,Miura C,et al.Tolerance of spermatogonia to oxidative stress is due to high levels of Zn and Cu/Zn superoxide dismutase[J].PLo S One,2011,6(2):e16938.
[12]Chung JY,Kim HJ,Kim M.The protective effect of growth hormone on Cu/Zn superoxide dismutase-mutant motor neurons[J].BMC Neurosci,2015,16:1.
[13]冯亚静,高利华,段丽菊,等.锌对染氟大鼠生精细胞Bcl-2和Bax蛋白表达的影响[J].郑州大学学报(医学版),2011,46(1):48-50.
[14]Johnson AL.Intracellular mechanisms regulating cell survival in ovarian follicles[J].Anim Reprod Sci,2003,78(34):185-188.
[15]王飞清,李艳菊,王宁,等.氧化应激-炎性因子对氟中毒卵巢早衰的影响[J].现代预防医学,2017,44(12):2296-2299.
[16]Rachel PU,John MD.Vitamin D in pregnancy:current concepts[J].Curr Opin Obstet Gynecol,2012,24(2):57-64.
[17]李学会,夏曙华,王世君,等.多种营养元素和强化大豆对氟中毒雌鼠性腺轴内分泌功能的作用[J].中华地方病学杂志,2015,34(9):660-665.
[2]王世君,夏曙华,余司文,等.煤氟污染的粮食对雌鼠动情周期的影响[J].中华地方病学杂志,2014,33(2):138-140.
[3]王飞清,余司文,夏曙华,等.燃煤型氟中毒对雌鼠卵巢生殖功能影响[J].中国公共卫生,2015,31(1):76-79.
[4]邹文兵,夏曙华,徐静峰,等.燃煤型氟中毒对雌性大鼠卵巢颗粒细胞的影响[J].环境与健康杂志,2014,31(2):124-127.
[5]李学会,夏曙华,王世君,等.复合营养素和蛋白质协同拮抗氟中毒对雌性大鼠卵巢损伤的动态观察[J].环境与健康杂志,2014,31(12):1055-1059.
[6]余司文,王飞清,夏曙华,等.多种营养素干预燃煤型氟中毒对雌鼠氟斑牙及骨氟和尿氟的影响[J].环境与健康杂志,2017,34(6):477-480.
[7]刘洋,韩伦英,何川,等.不同性别氟中毒大鼠模型的比较研究[J].中国比较医学杂志,2018,28(1):33-37.
[8]Fan B,Yu Y,Zhang Y.PI3K-Akt1 expression and its significance in liver tissues with chronic fluorosis[J].Int J Clin Exp Pathol,2015,8(2):1226-1236.
[9]Sahin N,Onderci M,Sahin K,et al.Magnesium proteinate is more protective than magnesium oxide in heat-stressed quail[J].J Nutr,2005,135(7):1732-1737.
[10]刘永祥,呙于明.饲粮镁对肉仔鸡生长、组织镁浓度及抗氧化功能的影响[J].中国畜牧杂志,2007,43(19):29-33.
[11]Celino FT,Yamaguchi S,Miura C,et al.Tolerance of spermatogonia to oxidative stress is due to high levels of Zn and Cu/Zn superoxide dismutase[J].PLo S One,2011,6(2):e16938.
[12]Chung JY,Kim HJ,Kim M.The protective effect of growth hormone on Cu/Zn superoxide dismutase-mutant motor neurons[J].BMC Neurosci,2015,16:1.
[13]冯亚静,高利华,段丽菊,等.锌对染氟大鼠生精细胞Bcl-2和Bax蛋白表达的影响[J].郑州大学学报(医学版),2011,46(1):48-50.
[14]Johnson AL.Intracellular mechanisms regulating cell survival in ovarian follicles[J].Anim Reprod Sci,2003,78(34):185-188.
[15]王飞清,李艳菊,王宁,等.氧化应激-炎性因子对氟中毒卵巢早衰的影响[J].现代预防医学,2017,44(12):2296-2299.
[16]Rachel PU,John MD.Vitamin D in pregnancy:current concepts[J].Curr Opin Obstet Gynecol,2012,24(2):57-64.
[17]李学会,夏曙华,王世君,等.多种营养元素和强化大豆对氟中毒雌鼠性腺轴内分泌功能的作用[J].中华地方病学杂志,2015,34(9):660-665.
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