脂联素(AdipoQ),脂肪细胞分泌的脂肪因子,据报道,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然难以捉摸。在这项研究中,我们研究了AdipoQ对增殖的影响,凋亡,相关激素的分泌(FSH,LH,TSH,GH,PRL和ACTH)和相关基因(FSHβ,LHβ,GnRHR,TSHβ,GH,通过使用实时荧光定量PCR(RT-qPCR)对鸡原代腺垂体细胞进行PRL和ACTH),细胞计数试剂盒-8(CCK-8),流式细胞术,酶联免疫吸附测定(ELISA)和蛋白质印迹(WB)测定。我们的结果表明,AdipoQ促进鸡原代腺垂体细胞的增殖,上调增殖相关基因CDK1、PCNA、CCND1和P21(P<0.05),CDK1和PCNA蛋白表达增加(P<0.05)。此外,AdipoQ抑制鸡原代腺垂体细胞凋亡,导致促凋亡基因Caspase3,Fas,和FasLmRNA表达,Caspase3蛋白表达下降(P<0.05)。此外,抗凋亡基因Bcl2mRNA和蛋白表达上调(P<0.05)。此外,AdipoQ抑制FSH的分泌,LH,TSH,GH,PRL,和ACTH(P<0.05),相关基因的mRNA表达水平(P<0.05)。用AdipoRon(AdipoQ的合成替代品)处理和与靶向AdipoQ受体1/2(AdipoR1/2)的RNA干扰共同处理对FSH的分泌没有影响,LH,TSH,GH,PRL,ACTH,以及相关基因的mRNA表达水平。这表明AdipoQ对激素分泌和相关基因表达的调节是由AdipoR1/2信号轴介导的。重要的是,我们进一步证明了AdipoQ对FSH的作用机制,LH,TSH和GH的分泌是通过AMPK信号通路实现的。总之,我们已经透露,首次研究了AdipoQ调节鸡原代腺垂体细胞激素分泌的分子机制。
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.