槐提取物和维生素C组合物对紫外照射诱导的HaCaT细胞光损伤的保护作用

孙璇; 李萍; 孙静; 郑嘉妮; 余丹阳; 张艳美; 陈朋

doi:10.13386/j.issn1002-0306.2023010057

槐提取物和维生素C组合物对紫外照射诱导的HaCaT细胞光损伤的保护作用

Protective Effect of Sophora japonica L. Extract and Vitamin C Composition on Photodamage of HaCaT Cells Induced by Ultraviolet Irradiation

摘要

摘要: 本研究旨在探讨槐提取物和维生素C（Vitamin C，V_C）对紫外线照射引起的HaCaT细胞光损伤的保护作用。通过体外培养HaCaT细胞并将其分为不同组别：对照组（未接受紫外辐照，无槐提取物、V_C或二者组合物处理）、紫外线辐射组、125 μg/mL槐提取物（接受紫外辐照及槐提取物处理）、125 μg/mL维生素C（接受紫外辐照及维生素C处理）以及125 μg/mL组合物组（接受紫外辐照及组合物处理，槐提取物和V_C质量比为1:1）。采用荧光染色和酶标仪检测技术来评估槐提取物、V_C及二者组合物对紫外照射引起的HaCaT细胞产生的活性氧物质（ROS）和线粒体ROS的抑制率。利用免疫荧光染色和酶联免疫吸附法检测槐提取物、V_C和组合物对紫外照射引起的HaCaT细胞中环丁烷嘧啶二聚体（Cyclobutane pyrimidine dimers，CPDs）和细胞炎症因子IL-6的表达水平。结果显示，槐提取物、V_C和组合物对UVA处理后HaCaT细胞生成的总ROS抑制率分别为38.23%±9.23%、27.20%±6.87%及64.13%±4.08%，与单独使用槐提取物或V_C处理相比，组合物显著提高总ROS抑制率（P<0.01）。槐提取物、V_C和组合物对线粒体ROS（Mitochondrial DNA，mtROS）的抑制率分别为50.29%±4.92%、38.81%±8.66%及84.74%±3.68%，与单独使用槐提取物或V_C处理相比，组合物极显著提高mtROS抑制率（P<0.01）。此外，在UVB照射后，槐提取物、V_C及组合物可极显著减少CPDs的生成（P<0.01），这种效应在组合物组效果更为显著（与槐提取物组比较P<0.05；与V_C组比较P<0.01）。同时，槐提取物和组合物显著降低UVA照射后炎症因子IL-6的分泌（P<0.01；P<0.05），表明槐提取物及组合物具有抗炎作用。本研究结果表明槐提取物和维生素C组合物能减轻紫外线引起的氧化应激损伤和炎症反应，为未来体内研究和开发具有健康益处的槐提取物和V_C组合物提供实验支持。

Abstract: This study aims to investigate the protective effects of the extract of Sophora japonica L. (Sophora extract) and vitamin C (V_C) on UV-induced damage in HaCaT cells. HaCaT cells were cultured in vitro and divided into different groups: Control group (without UV irradiation or treatment with Sophora extract, V_C, or their combination), UV radiation group, 125 μg/mL Sophora extract group (with UV irradiation and treatment with Sophora extract), 125 μg/mL vitamin C group (with UV irradiation and treatment with V_C), and 125 μg/mL combination group (with UV irradiation, Sophora extract and V_C at a mass ratio of 1:1). The inhibitory effects of Sophora extract, V_C, and their combination on reactive oxygen species (ROS) production and mitochondrial ROS in UV-irradiated HaCaT cells were measured using fluorescence staining and enzyme-linked immunosorbent assay. Immunofluorescence staining and enzyme-linked immunosorbent assay were also employed to evaluate the expression levels of cyclobutane pyrimidine dimers (CPDs) and the inflammatory cytokine IL-6 in UV-irradiated HaCaT cells treated with Sophora extract, V_C, or their combination. The results showed that the total ROS inhibition rates of Sophora extract, V_C, and the combination in UVA-treated HaCaT cells were 38.23%±9.23%, 27.20%±6.87%, and 64.13%±4.08%, respectively. The combination significantly enhanced the total ROS inhibition rate compared to the individual treatments (P<0.01). The inhibition rates of mitochondrial ROS (mtROS) by Sophora extract, V_C, and the combination were 50.29%±4.92%, 38.81%±8.66%, and 84.74%±3.68%, respectively. The combination significantly increased the mtROS inhibition rate compared to the individual treatments (P<0.01). After UVB irradiation, the CPDs content in HaCaT cells increased, and Sophora extract, V_C, and their combination significantly reduced CPDs formation (P<0.01), with the combination showing a more pronounced effect compared to the Sophora extract group (P<0.05) and the V_C group (P<0.01) alone. Furthermore, Sophora extract and the combination significantly reduced the secretion of the inflammatory cytokine IL-6 in HaCaT cells after UVA irradiation (P<0.01, P<0.05), indicating an anti-inflammatory capability. The results demonstrated that the combination of Sophora extract and Vitamin C could alleviate oxidative stress damage and inflammatory responses caused by UV radiation and provided experimental support for future in vivo studies and the development and utilization of health-promoting Sophora extract and V_C combinations.