Abstract：
UNASSIGNED: Psoriasis is characterized by inflammation and hyperproliferation of epidermal keratinocytes. Cycloastragenol (CAG) is an active molecule of Astragalus membranaceus that potentially plays a repressive role in psoriasis. Activated cell autophagy is an effective pathway for alleviating psoriasis progression. Thus, we investigated the role of CAG in the proliferation and autophagy of interleukin (IL)-22-stimulated keratinocytes.
UNASSIGNED: A psoriasis model was established by stimulating HaCaT cells with IL-22. Gene or protein expression levels were measured by qRT-PCR or western blot. Autophagy flux was observed with mRFP-GFP-LC3 adenovirus transfection assay under confocal microscopy. Stanniocalcin-1 (STC1) secretion levels were determined using ELISA kits. The apoptosis rate was assessed using flow cytometry. Interactions between miR-145 and STC1 or STC1 and Notch1 were validated by luciferase reporter gene assays, RIP, and Co-IP assays.
UNASSIGNED: CAG repressed cell proliferation and promoted apoptosis and autophagy in IL-22-stimulated HaCaT cells. Additionally, CAG promoted autophagy by enhancing miR-145. STC1 silencing ameliorated autophagy repression in IL-22-treated HaCaT cells. Moreover, miR-145 negatively regulated STC1, and STC1 was found to activate Notch1. Lastly, STC1 overexpression reversed CAG-promoted autophagy.
UNASSIGNED: CAG alleviated keratinocyte hyperproliferation through autophagy enhancement via regulating the miR-145/STC1/Notch1 axis in psoriasis.
UNASSIGNED: A psoriasis model was established by stimulating HaCaT cells with IL-22. Gene or protein expression levels were measured by qRT-PCR or western blot. Autophagy flux was observed with mRFP-GFP-LC3 adenovirus transfection assay under confocal microscopy. Stanniocalcin-1 (STC1) secretion levels were determined using ELISA kits. The apoptosis rate was assessed using flow cytometry. Interactions between miR-145 and STC1 or STC1 and Notch1 were validated by luciferase reporter gene assays, RIP, and Co-IP assays.
UNASSIGNED: CAG repressed cell proliferation and promoted apoptosis and autophagy in IL-22-stimulated HaCaT cells. Additionally, CAG promoted autophagy by enhancing miR-145. STC1 silencing ameliorated autophagy repression in IL-22-treated HaCaT cells. Moreover, miR-145 negatively regulated STC1, and STC1 was found to activate Notch1. Lastly, STC1 overexpression reversed CAG-promoted autophagy.
UNASSIGNED: CAG alleviated keratinocyte hyperproliferation through autophagy enhancement via regulating the miR-145/STC1/Notch1 axis in psoriasis.
摘要：
■银屑病的特征在于表皮角质形成细胞的炎症和过度增殖。环黄芪醇（CAG）是黄芪的活性分子，可能在牛皮癣中起抑制作用。激活的细胞自噬是缓解银屑病进展的有效途径。因此，我们研究了CAG在白细胞介素（IL）-22刺激的角质形成细胞的增殖和自噬中的作用。
■用IL-22刺激HaCaT细胞建立银屑病模型。通过qRT-PCR或蛋白质印迹测量基因或蛋白质表达水平。在共聚焦显微镜下用mRFP-GFP-LC3腺病毒转染测定观察自噬通量。使用ELISA试剂盒测定斯妥卡素-1(STC1)分泌水平。使用流式细胞术评估细胞凋亡率。miR-145和STC1或STC1和Notch1之间的相互作用通过荧光素酶报告基因检测进行验证。RIP,和Co-IP测定。
■CAG抑制IL-22刺激的HaCaT细胞增殖，促进细胞凋亡和自噬。此外,CAG通过增强miR-145促进自噬。STC1沉默改善了IL-22处理的HaCaT细胞中的自噬抑制。此外，miR-145负调控STC1，发现STC1激活Notch1。最后,STC1过表达可逆转CAG促进自噬。
■CAG通过调节miR-145/STC1/Notch1轴增强自噬减轻银屑病角质形成细胞过度增殖。
■用IL-22刺激HaCaT细胞建立银屑病模型。通过qRT-PCR或蛋白质印迹测量基因或蛋白质表达水平。在共聚焦显微镜下用mRFP-GFP-LC3腺病毒转染测定观察自噬通量。使用ELISA试剂盒测定斯妥卡素-1(STC1)分泌水平。使用流式细胞术评估细胞凋亡率。miR-145和STC1或STC1和Notch1之间的相互作用通过荧光素酶报告基因检测进行验证。RIP,和Co-IP测定。
■CAG抑制IL-22刺激的HaCaT细胞增殖，促进细胞凋亡和自噬。此外,CAG通过增强miR-145促进自噬。STC1沉默改善了IL-22处理的HaCaT细胞中的自噬抑制。此外，miR-145负调控STC1，发现STC1激活Notch1。最后,STC1过表达可逆转CAG促进自噬。
■CAG通过调节miR-145/STC1/Notch1轴增强自噬减轻银屑病角质形成细胞过度增殖。
