Isozymes of glutathione peroxidase in cow ovarian tissue under different physiological and pathological states of the gonad

M. М. Akymyshyn, N. Kuzmina, D. Ostapiv
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Abstract

The article presents a study of glutathione peroxidase (GPO) isozymes content in ovarian tissue under various physiological and pathological states of cow gonads. The ovarian tissue of 12 cows in different physiological conditions were selected for research, from which homogenates were prepared. The content of isozymes was determined in the supernatant after centrifugation by electrophoresis. After electrophoresis in a 7.5 % polyacrylamide gel and specific staining of the plates according to Lin C. L. and Weydert C. J., 5 bands of catalytically active isozymes of the enzyme (GPO1, GPO2, GPO3, GPO4, and GPO5) were established. They differed in electrophoretic mobility, intensity, and area of band staining. It has been proven that depending on the physiological state of the ovaries, the content of individual isozymes in the gonad tissue changed. In particular, it was found that the content of GPO1 for “late corpus luteum” and “follicular growth” was almost the same, and was in the range of 14.2–15.6 %, and for “early corpus luteum” it was 8.7–10.1 % lower (Р < 0.01). The content of GPO2 was high in “early corpus luteum” (19.9 ± 6.42 %), and in “late corpus luteum” and “follicular growth” it was lower on 11.0–13.2 % (Р < 0.01). The content of GPO3 during “follicular growth” and “early corpus luteum” was within 50.1–51.2 %, and during “late” it decreased on 4.5–5.6 %. According to changes in the physiological state of the ovary in the tissue of “late corpus luteum”, the content of GPO4 was the highest (26.0 ± 2.40 %), and under “follicular growth” and “early corpus luteum” it was lower on 10.2 (Р < 0.05) and 12.9 % (Р < 0.01), respectively. A high content of GPO5 was established in the ovarian tissue of “follicular growth” (10.8 ± 2.00 %), which tended to decrease by 2.8 % and by 4.8 %, respectively, in “early” and “late corpus luteum”. For ovarian tissue with hypofunction, compared to physiological conditions, a probable decrease in the content of the following isozymes was registered: GPO1 by 1.2–11.3 % (Р < 0.01), GPO4 – 6.8–19.7 % (Р < 0 .01) and GPO5 – 2.7–7.5 % (Р < 0.01), while the content of GPO3, on the contrary, increased by 23.6–29.2 % (Р < 0.05–0.01). The study of the dependence of the content of enzyme isozymes on different conditions of the gonad showed that the correlation ratio for the content of GPO isozymes in ovarian tissue under physiological conditions and hypofunction: medium strength was negative for GPO2 and GPO3 (η = 0.473 and 0.530), positive for GPO5 (η = 0.481 ), and strongly positive for GPO1 and GPO4 (η = 0.740 and 0.746).
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不同性腺生理病理状态下奶牛卵巢组织谷胱甘肽过氧化物酶同工酶的研究
本文研究了不同生理病理状态下奶牛卵巢组织谷胱甘肽过氧化物酶(GPO)同工酶含量的变化。选取12头奶牛不同生理状态下的卵巢组织进行研究,制备匀浆。离心后电泳测定上清中同工酶的含量。根据Lin C. L.和Weydert C. J.的方法,经7.5%聚丙烯酰胺凝胶电泳和特异性染色后,建立了GPO1、GPO2、GPO3、GPO4和GPO5 5条催化活性同工酶带。它们在电泳迁移率、强度和条带染色面积上存在差异。已经证明,根据卵巢的生理状态,性腺组织中单个同工酶的含量会发生变化。特别是发现“晚期黄体”和“卵泡生长”的GPO1含量几乎相同,在14.2 ~ 15.6%的范围内,而“早期黄体”的GPO1含量低8.7 ~ 10.1% (Р < 0.01)。GPO2在“早期黄体”中含量较高(19.9±6.42%),在“晚期黄体”和“卵泡生长”中含量较低(11.0 ~ 13.2%)(Р < 0.01)。在“卵泡生长”和“早期黄体”期间,GPO3含量在50.1 - 51.2%之间,在“晚期”期间,GPO3含量下降了4.5 - 5.6%。从卵巢生理状态变化来看,“晚期黄体”组织中GPO4含量最高(26.0±2.40%),“卵泡生长”和“早期黄体”组织中GPO4含量较低,分别为10.2 (Р < 0.05)和12.9% (Р < 0.01)。GPO5在卵巢“卵泡生长”组织中含量较高(10.8±2.00 %),在黄体“早期”和“晚期”分别有下降2.8%和4.8%的趋势。与生理状态相比,卵巢功能减退组织中GPO1、GPO4和GPO5同工酶的含量分别下降了1.2 ~ 11.3% (Р < 0.01)、6.8 ~ 19.7% (Р < 0.01)和2.7 ~ 7.5% (Р < 0.01),而GPO3的含量则增加了23.6 ~ 29.2% (Р < 0.05 ~ 0.01)。对不同性腺条件下卵巢组织GPO同工酶含量依赖性的研究表明,生理条件下卵巢组织GPO同工酶含量与功能低下的相关比值为:中等强度GPO2和GPO3为负(η = 0.473和0.530),GPO5为正(η = 0.481), GPO1和GPO4为强正(η = 0.740和0.746)。
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