紫外线预处理后的核黄素对淋巴细胞活力和凋亡率的影响

陈兴慧; 葛霜; 黄远帅; 杨璐; 汪德清

doi:10.12435/j.issn.2095-5227.2024.121

紫外线预处理后的核黄素对淋巴细胞活力和凋亡率的影响

Effects of UV- pretreated riboflavin on lymphocyte viability and apoptosis rate

摘要

摘要:
背景核黄素作为一种机体必需的维生素，其增强机体免疫功能的作用已被广泛研究，而经紫外线照射预处理后的核黄素对免疫细胞功能的影响却鲜有报道。目的 研究经紫外线预处理后的核黄素对淋巴细胞活力和凋亡率的影响，为核黄素的临床应用提供新的思路。
方法在体外将紫外线预处理后的核黄素与淋巴细胞共培养，探究了预处理核黄素浓度(0、50、100、200、400 μmol/L)、处理细胞浓度(2.5 × 10⁶/mL、5.0 × 10⁶/mL、1 × 10⁷/mL)以及预处理核黄素保存时间(Day1、Day3、Day7)三个参数对淋巴细胞凋亡率的影响。将不同浓度预处理核黄素(0、25、50、75、100、150、200 μmol/L)与淋巴细胞CD4⁺ T、CD8⁺ T共培养24 h，利用CCK-8增殖实验检测细胞活力。利用流式细胞技术检测不同浓度预处理核黄素(0、10、50、200、400 μmol/L)处理后CD4⁺ T、CD8⁺ T细胞凋亡率的变化，观察这两种细胞对处理的耐受性差异。
结果与对照组相比，紫外线预处理后的核黄素能够诱导淋巴细胞凋亡，且凋亡率与预处理核黄素浓度呈正相关(P＜0.01)，与处理细胞浓度(P＜0.01)和预处理核黄素保存时间(P＜0.05)呈负相关。当预处理核黄素浓度≤50 μmol/L时，CD4⁺ T、CD8⁺ T细胞活力基本不受影响，而＞50 μmol/L时，细胞活力总体呈现下降趋势(P＜0.05)。预处理核黄素浓度相同时，CD8⁺ T细胞凋亡率低于CD4⁺ T细胞(P＜0.01)。
结论紫外线预处理后的核黄素能够诱导淋巴细胞凋亡、降低CD4⁺ T和CD8⁺ T淋巴细胞活力，且CD8⁺ T细胞对处理的耐受性强于CD4⁺ T细胞。

Abstract:
Background As an essential vitamin, riboflavin has been extensively investigated for its role in boosting immune function. However, the effect of riboflavin on immune cell function after pretreatment by ultraviolet remains underreported.
Objective To investigate the impact of UV-pretreated riboflavin on lymphocyte viability and apoptosis rate, aiming to offer novel insights for the clinical application of riboflavin.
Methods The UV-pretreated riboflavin was co-cultured with lymphocytes in vitro to investigate the impact of UV-pretreated riboflavin concentration (0, 50, 100, 200, 400 μmol/L), cell concentration (2.5 × 10⁶/mL, 5.0 × 10⁶/mL, 1.0 × 10⁶/mL), and storage time of UV-pretreated riboflavin (Day 1, Day 3, Day 7) on the apoptosis rate of lymphocytes. Different concentrations of UV-pretreated riboflavin (0, 25, 50, 75, 100, 150, 200 μmol/L) were co-cultured with lymphocyte CD4 + T and CD8 + T for 24 hours, and the cell viability was assessed using the CCK-8 proliferation assay. The apoptosis rate of CD4⁺T and CD8⁺T cells was evaluated by flow cytometry after treatment with UV-pretreated riboflavin at various concentrations (0, 10, 25, 200, 400 μmol/L), aiming to investigate the disparity in tolerance between the two kinds of cells.
Results Compared to the control group, UV-pretreated riboflavin was found to induce lymphocyte apoptosis, with the rate of apoptosis showing a positive correlation with the concentration of UV-pretreated riboflavin (P<0.01). Additionally, there was a negative correlation observed the apoptosis rate and both the concentration of treated cells (P<0.01) and the storage time of UV-pretreated riboflavin (P<0.05). When the concentration of riboflavin was below 50 μmol/L, there was minimal impact on the activity of CD4⁺T and CD8⁺T cells; however, upon exceeding a concentration of 50 μmol/L, a noticeable decline in overall cell viability becomes evident (P<0.05). Additionally, when comparing equivalent riboflavin concentrations, a significantly lower apoptosis rate was observed in CD8⁺T cells compared to CD4⁺T cells (P<0.01).
Conclusion The UV-pretreated riboflavin can induce apoptosis in lymphocytes and suppress the activity of CD4⁺T and CD8⁺T cells. Notably, CD8⁺T cells exhibit higher tolerance towards this treatment compared to CD4⁺T cells.

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