College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China.
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang 453003, China.
Poult Sci. 2024 Oct;103(10):104137. doi: 10.1016/j.psj.2024.104137. Epub 2024 Jul 29.
Adiponectin (AdipoQ), an adipokine secreted by adipocytes, has been reported to exist widely in various cell types and tissues, including the adenohypophysis of chickens. However, the molecular mechanism by which AdipoQ regulates the function of chicken adenohypophysis remains elusive. In this study, we investigated the effects of AdipoQ on proliferation, apoptosis, secretion of related hormones (FSH, LH, TSH, GH, PRL and ACTH) and expression of related genes (FSHβ, LHβ, GnRHR, TSHβ, GH, PRL and ACTH) in primary adenohypophysis cells of chickens by using real-time fluorescent quantitative PCR (RT-qPCR), cell counting kit-8 (CCK-8), flow cytometry, enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) assays. Our results showed that AdipoQ promoted the proliferation of chicken primary adenohypophysis cells, up-regulated the mRNA expression of proliferation-related genes CDK1, PCNA, CCND1 and P21 (P < 0.05), as well as the increased protein expression of CDK1 and PCNA (P < 0.05). Furthermore, AdipoQ inhibited apoptosis of chicken primary adenohypophysis cells, resulting in down-regulation of pro-apoptotic genes Caspase3, Fas, and FasL mRNA expression, and decreased Caspase3 protein expression (P < 0.05). Moreover, there was an up-regulation of anti-apoptotic gene Bcl2 mRNA and protein expression (P < 0.05). Additionally, AdipoQ suppressed the secretion of FSH, LH, TSH, GH, PRL, and ACTH (P < 0.05), as well as the mRNA expression levels of related genes (P < 0.05). Treatment with AdipoRon (a synthetic substitute for AdipoQ) and co-treatment with RNA interference targeting AdipoQ receptors 1/2 (AdipoR1/2) had no effect on the secretion of FSH, LH, TSH, GH, PRL, and ACTH, as well as the mRNA expression levels of the related genes. This suggests that AdipoQ's regulation of hormone secretion and related gene expression is mediated by the AdipoR1/2 signaling axis. Importantly, we further demonstrated that the mechanism of AdipoQ on FSH, LH, TSH and GH secretion is realized through AMPK signaling pathway. In conclusion, we have revealed, for the first time the molecular mechanism by which AdipoQ regulates hormone secretion in chicken primary adenohypophysis cells.
脂联素（AdipoQ）是一种由脂肪细胞分泌的脂肪因子，已被报道广泛存在于各种细胞类型和组织中，包括鸡的腺垂体。然而，AdipoQ 调节鸡腺垂体功能的分子机制仍不清楚。在这项研究中，我们通过实时荧光定量 PCR（RT-qPCR）、细胞计数试剂盒-8（CCK-8）、流式细胞术、酶联免疫吸附测定（ELISA）和 Western blot（WB）实验研究了 AdipoQ 对鸡原代腺垂体细胞增殖、凋亡、相关激素（FSH、LH、TSH、GH、PRL 和 ACTH）分泌以及相关基因（FSHβ、LHβ、GnRHR、TSHβ、GH、PRL 和 ACTH）表达的影响。我们的结果表明，AdipoQ 促进了鸡原代腺垂体细胞的增殖，上调了增殖相关基因 CDK1、PCNA、CCND1 和 P21 的 mRNA 表达（P＜0.05），以及 CDK1 和 PCNA 的蛋白表达增加（P＜0.05）。此外，AdipoQ 抑制了鸡原代腺垂体细胞的凋亡，导致促凋亡基因 Caspase3、Fas 和 FasL 的 mRNA 表达下调，Caspase3 蛋白表达降低（P＜0.05）。此外，抗凋亡基因 Bcl2 的 mRNA 和蛋白表达上调（P＜0.05）。此外，AdipoQ 抑制了 FSH、LH、TSH、GH、PRL 和 ACTH 的分泌（P＜0.05），以及相关基因的 mRNA 表达水平（P＜0.05）。用 AdipoRon（AdipoQ 的合成替代物）处理和用 AdipoQ 受体 1/2（AdipoR1/2）的 RNA 干扰共同处理对 FSH、LH、TSH、GH、PRL 和 ACTH 的分泌以及相关基因的 mRNA 表达水平没有影响。这表明 AdipoQ 对激素分泌和相关基因表达的调节是通过 AdipoR1/2 信号轴介导的。重要的是，我们进一步证明了 AdipoQ 对 FSH、LH、TSH 和 GH 分泌的作用是通过 AMPK 信号通路实现的。总之，我们首次揭示了 AdipoQ 调节鸡原代腺垂体细胞激素分泌的分子机制。
