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The effects of apelin on IGF1/FSH-induced steroidogenesis, proliferation, Bax expression, and total antioxidant capacity in granulosa cells of buffalo ovarian follicles

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Abstract

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 (10−9 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 (10–9) 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 (10–8 and 10–9 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 10–6 and 10–8 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.

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Funding

This work was financially supported by grants from the Guangxi Science and Technology Major Project [GuiKe AA22068099-3], the Special Fund for Guiding Local Scientific and Technological Development by the Central Government (GuiKe ZY21195007), the Guangxi Natural Science Foundation (2018GXNSFDA050013) and the National Key Research and Development Program (2017YFE0113800) of China.

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Authors and Affiliations

  1. Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning, 530001, China

    Borhan Shokrollahi, Hai-Ying Zheng, Xiao-Ya Ma & Jiang-Hua Shang

  2. Department of Animal Science, Sanandaj Branch, Islamic Azad University, Sanandaj, Kurdistan, Iran

    Borhan Shokrollahi

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  1. Borhan Shokrollahi
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  2. Hai-Ying Zheng
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  4. Jiang-Hua Shang
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BS designed, conducted the study and data analysis, and also wrote the manuscript. HYZ and XYM helped with some lab work and JHS was the project leader. All authors contributed to the article and approved the submitted version.

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Correspondence to Jiang-Hua Shang.

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The Animal Ethics Committee at the Guangxi Buffalo Research Institute handled all aspects of the experiments and ensured they complied with animal research ethical regulations.

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Shokrollahi, B., Zheng, HY., Ma, XY. et al. The effects of apelin on IGF1/FSH-induced steroidogenesis, proliferation, Bax expression, and total antioxidant capacity in granulosa cells of buffalo ovarian follicles. Vet Res Commun 47, 1523–1533 (2023). https://doi.org/10.1007/s11259-023-10107-z

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