18β-Glycyrrhetinic acid induces human HaCaT keratinocytes apoptosis through ROS-mediated PI3K-Akt signaling pathway and ameliorates IMQ-induced psoriasis-like skin lesions in mice
18β-Glycyrrhetinic acid induces human HaCaT keratinocytes apoptosis through ROS-mediated PI3K-Akt signaling pathway and ameliorates IMQ-induced psoriasis-like skin lesions in mice
复制标题
18β-甘草次酸通过 ROS 介导的 PI3K-Akt 信号通路诱导人 HaCaT 角质形成细胞凋亡，并改善 IMQ 诱导的小鼠牛皮癣样皮肤病变
DOI：
10.1186/s40360-020-00419-0
复制
发表时间：
2020-06-03
影响因子：
2.9
通讯作者：
Yang, Mengjuan
中科院分区：
文献类型：
--
作者：
Gao, Jintao;Guo, Junfan;Yang, Mengjuan
背景：银屑病是一种慢性炎症性皮肤病，影响全球2-3%的人口。过度增殖的角质形成细胞被认为是炎症反应的放大因子，从而维持银屑病皮损的持久性。能够抑制角质形成细胞增殖或诱导细胞凋亡的药物对银屑病的治疗具有潜在的实用价值。18β-甘草次酸(GA)是甘草酸的活性代谢物，具有抗炎、抗菌、抗增殖等多种药理活性。本研究旨在观察赤霉素A对体外培养的人HaCaT角质形成细胞增殖和凋亡的影响，以及赤霉素A对咪喹莫特(Imiquimod，ImQ)诱导的银屑病样小鼠皮损的影响。分别用Annexin V-FITC/PI检测试剂盒和DCFH-DA探针检测细胞凋亡率和活性氧(ROS)。用caspase活性检测试剂盒检测caspase 9/3活性。用Western blotting检测Akt和p-Akt的蛋白水平。用IMQ诱导小鼠银屑病样皮肤病变。用苏木精-伊红(H&E)染色观察小鼠皮肤损伤的组织学变化。皮损严重程度根据银屑病面积严重程度指数(PASI)进行评分。结果：赤霉素A抑制HaCaT角质形成细胞活性，诱导细胞凋亡，并呈剂量依赖性。在GA存在的情况下，细胞内ROS水平显著升高。ROS抑制剂NAC可减弱GA介导的HaCaT角质形成细胞的生长抑制和凋亡。此外，GA可显著降低p-Akt蛋白水平，当GA和NAC共同处理细胞时，p-Akt蛋白水平可部分恢复。PI3K抑制剂LY294002可显著增强GA介导的细胞毒作用。结论：赤霉素A通过抑制PI3K-Akt信号通路抑制HaCaT角质形成细胞的增殖，诱导角质形成细胞凋亡，从而改善ImQ诱导的小鼠银屑病样皮肤病变。
Background: Psoriasis is a chronic inflammatory skin disease affecting 2-3% of the population worldwide. Hyperproliferative keratinocytes were thought to be an amplifier of inflammatory response, thereby sustaining persistence of psoriasis lesions. Agents with the ability to inhibit keratinocyte proliferation or induce apoptosis are potentially useful for psoriasis treatment. 18 beta-Glycyrrhetinic acid (GA), an active metabolite of glycyrrhizin, exhibits diverse pharmacological activities, including anti-inflammatory, anti-bacteria and anti-proliferation. The current study aims to evaluate the effects of GA on the proliferation and apoptosis of human HaCaT keratinocytes in vitro and investigate the effects of GA on the skin lesions of imiquimod (IMQ)-induced psoriasis-like mouse model in vivo.Methods: Cell viability was assayed by CCK-8. Flow cytometry was performed to measure apoptosis and reactive oxygen species (ROS), with Annexin V-FITC/PI detection kit and DCFH-DA probe respectively. Caspase 9/3 activities were measured using caspase activity assay kits. The protein levels of Akt and p-Akt were determined using Western blotting. IMQ was applied to induce psoriasis-like skin lesions in mice. The histological change in mouse skin lesions was detected using hematoxylin and eosin (H&E) staining. The severity of skin lesions was scored based on Psoriasis Area Severity Index (PASI). RT-PCR was employed to examine the relative expression of TNF-alpha, IL-22 and IL-17A in mouse skin lesions.Results: GA decreased HaCaT keratinocytes viability and induced cell apoptosis in a dose-dependent manner. In the presence of GA, intracellular ROS levels were significantly elevated. NAC, a ROS inhibitor, attenuated GA-mediated HaCaT keratinocytes growth inhibition and apoptosis. In addition, GA treatment remarkably decreased p-Akt protein level, which could be restored partially when cells were co-treated with GA and NAC. LY294002 (a PI3K inhibitor) treatment significantly enhanced GA-mediated cytotoxicity. Moreover, GA ameliorated IMQ-induced psoriasis-like skin lesions in mice.Conclusions: GA inhibits proliferation and induces apoptosis in HaCaT keratinocytes through ROS-mediated inhibition of PI3K-Akt signaling pathway, and ameliorates IMQ-induced psoriasis-like skin lesions in mice.