Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China.
Department of Cell Biology, Jinan University, Guangzhou, China.
J Cell Mol Med. 2016 May;20(5):874-90. doi: 10.1111/jcmm.12772. Epub 2016 Jan 22.
Pituitary adenylate cyclase-activating polypeptide (PACAP) is a structurally endogenous peptide with many biological roles. However, little is known about its presence or effects in human adipose-derived stem cells (hADSCs). In this study, the expression of PACAP type I receptor (PAC1R) was first confirmed in hADSCs. Maxadilan, a specific agonist of PAC1R, could increase hADSC proliferation as determined by Cell Counting Kit-8 and cell cycle analysis and promote migration as shown in wound-healing assays. Maxadilan also showed anti-apoptotic activity in hADSCs against serum withdrawal-induced apoptosis based on Annexin V/propidium iodide analysis and mitochondrial membrane potential assays. The anti-apoptotic effects of maxadilan correlated with the down-regulation of Cleaved Caspase 3 and Caspase 9 as well as up-regulation of Bcl-2. The chemical neural differentiation potential could be enhanced by maxadilan as indicated through quantitative PCR, Western blot and cell morphology analysis. Moreover, cytokine neural redifferentiation of hADSCs treated with maxadilan acquired stronger neuron-like functions with higher voltage-dependent tetrodotoxin-sensitive sodium currents, higher outward potassium currents and partial electrical impulses as determined using whole-cell patch clamp recordings. Maxadilan up-regulated the Wnt/β-catenin signalling pathway associated with dimer-dependent activity of PAC1R, promoting cell viability that was inhibited by XAV939, and it also activated the protein kinase A (PKA) signalling pathway associated with ligand-dependent activity of PAC1R, enhancing cell viability and neural differentiation potential that was inhibited by H-89. In summary, these results demonstrated that PAC1R is present in hADSCs, and maxadilan could enhance hADSC viability and neural differentiation potential in neural differentiation medium.
垂体腺苷酸环化酶激活肽（PACAP）是一种具有多种生物学作用的结构内源性肽。然而，人们对其在人脂肪来源干细胞（hADSCs）中的存在或作用知之甚少。在这项研究中，首次证实了 PACAP 型 I 受体（PAC1R）在 hADSCs 中的表达。Maxadilan 是 PAC1R 的特异性激动剂，可通过 Cell Counting Kit-8 和细胞周期分析确定增加 hADSC 增殖，并在划痕愈合试验中显示促进迁移。Maxadilan 还显示出抗 hADSC 凋亡活性，可抵抗血清剥夺诱导的凋亡，基于 Annexin V/碘化丙啶分析和线粒体膜电位测定。Maxadilan 的抗凋亡作用与 Cleaved Caspase 3 和 Caspase 9 的下调以及 Bcl-2 的上调相关。Maxadilan 可增强化学神经分化潜能，如通过定量 PCR、Western blot 和细胞形态分析所示。此外，用 maxadilan 处理的 hADSCs 的细胞因子神经再分化获得了更强的神经元样功能，具有更高的电压依赖性河豚毒素敏感钠电流、更高的外向钾电流和部分电脉冲，通过全细胞膜片钳记录确定。Maxadilan 上调了与 PAC1R 的二聚体依赖性活性相关的 Wnt/β-catenin 信号通路，促进了被 XAV939 抑制的细胞活力，并且还激活了与 PAC1R 的配体依赖性活性相关的蛋白激酶 A（PKA）信号通路，增强了被 H-89 抑制的细胞活力和神经分化潜能。总之，这些结果表明 PAC1R 存在于 hADSCs 中，maxadilan 可在神经分化培养基中增强 hADSC 的活力和神经分化潜能。
