摘要
目的:观察蒿秦化斑方对HaCat细胞紫外线B(UVB)急性损伤模型细胞黏附分子及PI3K/AKT/GSK-3β通路的影响,探讨该方治疗光敏感性皮肤病的作用机制。方法:采用300 mJ/cm2 UVB建立HaCat细胞急性光损伤模型,使用不同剂量蒿秦化斑方干预,分为空白对照组、UVB模型组、及蒿秦化斑方高、中、低剂量组。采用RT-PCR法检测细胞黏附分子ICAM-1、ELAM-1、VCAM-1相对表达量,采用蛋白质免疫印迹法分析PI3K/AKT/GSK-3β通路相关节点p-AKT(Ser-473)、p-AKT(Thr-308)、GSK-3β、PI3K相对表达量。结果:UVB急性光损伤可有效减少HaCat细胞ICAM-1的表达(P<0.01),对VCAM-1及ELAM-1表达影响不显著;蒿秦化斑方高剂量组能有效抑制HaCat细胞3种细胞黏附分子的表达(P<0.05);蒿秦化斑方中剂量组能有效抑制HaCat细胞内皮细胞白细胞黏附分子-1(ELAM-1)及血管细胞黏附分子-1(VCAM-1)的表达(P<0.01)。UVB急性光损伤对HaCat细胞AKT磷酸化、PI3K表达影响不显著,蒿秦化斑方低剂量处理对HaCat细胞AKT磷酸化具有促进作用(P<0.05)。UVB急性光损伤使HaCat细胞糖原合成酶激酶-3β(GSK-3β)表达水平升高(P<0.01),蒿秦化斑方高、中、低剂量处理均对HaCat细胞GSK-3β表达水平有所抑制(P<0.01)。结论:蒿秦化斑方能降低由UVB急性损伤造成的HaCat细胞ICAM-1、ELAM-1、VCAM-1等黏附分子的表达进而缓解局部炎症和迟发型超敏反应的发生,并且能够激活HaCat细胞PI3K/AKT/GSK-3β通路,在UVB造成的急性光损伤中起到抗氧化、抗凋亡和降低炎症反应的作用。
Objective:To observe the effects of Haoqin-Huaban on the relative adhesion factors and the PI3K/AKT/GSK-3βpathway of an acute UVB light HaCat-cell injury model in order to further explore the mechanism of the treatment for photosensitive skin disorders.Methods:UVB light(300 mJ/cm2)was used to establish an acute light HaCat-cell injury model.The intervention was performed by using Haoqin-Huaban in various concentrations.The groups were divided into:a blank control group(C);an UVB model group(M);a high Haoqin-Huaban dosage group(G);medium Haoqin-Huaban dosage group(Z);low Haoqin-Huaban dosage group(D).Real-time PCR was conducted to detect the relative expressions of 3 cell adhesion molecules(CAMs),i.e.,ICAM-1,ELAM-1 and VCAM-1.Western Blotting was implemented to analyze the relative expressions of p-AKT(Ser-473),p-AKT(Thr-308),GSK-3β,and PI3K,which were all related nodes of the PI3K/AKT/GSK-3βpathway.Western Blot method was used to analyze the relative expression of p-AKT(Ser-473),p-AKT(Thr-308),GSK-3βand PI3K related nodes in the PI3K/AKT/GSK-3βpathway.Results:UVB acute light injury reduced ICAM-1 expression in the HaCat cell model and had no significant effect on the expressions of VCAM-1 and ELAM-1(P<0.01).The expressions of the three CAMs in the HaCat cell model were effectively inhibited in the G group(P<0.05),while those of ELAM-1 and VCAM-1 were considerably impeded in the Z group(P<0.01).It was also found that acute UVB light injury had no significant effect on AKT phosphorylation and PI3K expression in the HaCat cell model.The low-concentration treatment of Haoqin Huaban Forumla can promote the phosphorylation of AKT in HaCat cells(P<0.05).Acute UVB light damage increased the GSK-3βexpression level of HaCat cells(P<0.01).The high,medium and low concentration treatments of Haoqin Huaban Formula inhibited the GSK-3βexpression level of HaCat cells(P<0.01).Conclusion:Haoqin-Huaban Formula can reduce the expression of adhesion molecules such as ICAM-1,ELAM-1 and VCAM-1 in HaCat cells caused by acute UVB damage,thereby alleviating the occurrence of local inflammation and delayed-type hypersensitivity,and can activate HaCat cells PI3K/AKT/GSK-3βpathway and plays a role in anti-oxidation,anti-apoptosis and reducing inflammation in the acute light damage caused by UVB.
作者
王思艺
孙丽蕴
WANG Siyi;SUN Liyun(Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China)
出处
《世界中医药》
CAS
2021年第19期2876-2880,共5页
World Chinese Medicine
基金
国家自然科学基金面上项目(81774309)
北京中医医院顺义医院院级课题(SYYJ-201806)。
