Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning, 530001, China.
Department of Animal Science, Sanandaj Branch, Islamic Azad University, Sanandaj, Kurdistan, Iran.
Vet Res Commun. 2023 Sep;47(3):1523-1533. doi: 10.1007/s11259-023-10107-z. Epub 2023 Apr 10.
Apelin (APLN) was believed to be an adipokine secreted from adipose tissue. However, studies demonstrate that it is a pleiotropic peptide and has several effects on the female reproductive system. In this study, We examined the effects of different doses of IGF1 and FSH in the presence of APLN-13 on the production of progesterone in buffalo ovary granulosa cells. Furthermore, different doses of APLN isoforms (APLN-13 and APLN-17) were tested on proliferation, Bax protein expression, and antioxidant capacity in the same cells. Granulosa cells of buffalo ovaries were cultured in the presence of different doses of IGF1 and FSH with or without APLN-13 (10 M) to evaluate its effect on the secretion of progesterone tested by ELISA assay. The WST-1 method was used to survey the effect of APLN on granulosa cell proliferation and cytotoxicity. In addition, the antioxidant capacity of the cells in the presence of APLN was assessed using the FRAP method. mRNA and Bax protein levels were measured in granulosa cells treated with APLN using real-time PCR and western blot techniques. APLN-13 (10) stimulated the effect of IGF1 on the production of progesterone, and its levels were affected by APLN-13 dose-dependently. However, it did not significantly stimulate the effect of FSH on the secretion of progesterone. APLN-13 (all doses) and APLN-17 (10 and 10 M) improved the proliferation of granulosa cells. Moreover, preincubation of the cells for an hour by APLN receptor antagonist (ML221, 10 µM) did not significantly affect the proliferation of cells induced by APLN. Neither APLN-13 nor APLN-17 were not cytotoxic for the cells compared to the control treatment. APLN-13 at the doses of 10 and 10 M substantially up and down-regulated Bax protein expression; however, such effects were not observed when the cells were preincubated with ML221. In addition, APLN-17 did not influence the expression amount of Bax. Furthermore, both APLN-13 and -17 improved the total antioxidant capacity of the ovarian granulosa cells, but such effects were not seen when the cells were preincubated with ML221. According to these results, APLN enhanced the steroidogenesis induced by IGF1 but did not affect the steroidogenesis induced by FSH. APLN also enhanced the cell proliferation and antioxidant capacity of buffalo ovaries follicular granulosa cells; however, its effect on Bax expression was different.
Apelin (APLN) 被认为是一种脂肪组织分泌的脂肪因子。然而，研究表明它是一种多效肽，对女性生殖系统有多种影响。在这项研究中，我们研究了不同剂量的 IGF1 和 FSH 在 APLN-13 存在下对水牛卵巢颗粒细胞中孕酮产生的影响。此外，还测试了不同剂量的 APLN 同工型（APLN-13 和 APLN-17）对同一细胞增殖、Bax 蛋白表达和抗氧化能力的影响。在不同剂量的 IGF1 和 FSH 存在下培养水牛卵巢颗粒细胞，并添加或不添加 APLN-13（10μM），通过 ELISA 测定评估其对孕酮分泌的影响。WST-1 法用于研究 APLN 对颗粒细胞增殖和细胞毒性的影响。此外，还使用 FRAP 法评估了 APLN 存在下细胞的抗氧化能力。使用实时 PCR 和 Western blot 技术测量用 APLN 处理的颗粒细胞中的 APLN mRNA 和 Bax 蛋白水平。APLN-13（10）刺激 IGF1 对孕酮产生的作用，其水平受 APLN-13 剂量依赖性影响。然而，它并没有显著刺激 FSH 对孕酮分泌的作用。APLN-13（所有剂量）和 APLN-17（10 和 10μM）均能促进颗粒细胞的增殖。此外，用 APLN 受体拮抗剂（ML221，10μM）预孵育细胞 1 小时，对 APLN 诱导的细胞增殖没有显著影响。与对照处理相比，APLN-13 和 APLN-17 对细胞均无细胞毒性。APLN-13 在 10 和 10μM 的剂量下，显著上调和下调 Bax 蛋白表达；然而，当细胞用 ML221 预孵育时，未观察到这种作用。此外，APLN-17 不影响 Bax 的表达量。此外，APLN-13 和 APLN-17 均能提高卵巢颗粒细胞的总抗氧化能力，但当细胞用 ML221 预孵育时，未见这种作用。根据这些结果，APLN 增强了 IGF1 诱导的类固醇生成，但不影响 FSH 诱导的类固醇生成。APLN 还增强了水牛卵巢卵泡颗粒细胞的细胞增殖和抗氧化能力；然而，其对 Bax 表达的影响不同。
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