玉米黄素对衣霉素诱导的SH-SY5Y细胞周期、自噬、凋亡的保护作用

doi:10.7506/spkx1002-6630-20210325-318

食品科学 ›› 2022, Vol. 43 ›› Issue (7): 112-119.doi: 10.7506/spkx1002-6630-20210325-318

玉米黄素对衣霉素诱导的SH-SY5Y细胞周期、自噬、凋亡的保护作用

章丽娜，李梦洁，商迎辉，刘芸如，黄汉昌，劳凤学

（北京市生物活性物质和功能食品重点实验室，北京联合大学功能因子与脑科学研究院，北京 100191）

出版日期:2022-04-15 发布日期:2022-04-26
基金资助:
国家自然科学基金面上项目（31471587）；北京联合大学科研项目（ZK40201902；JZ10202001；XP202008）；北京联合大学研究生科研创新资助项目（YZ2020K001）

Protective Effect of Zeaxanthin on Tunicamycin-Induced Cell Cycle Arrest, Autophagy and Apoptosis of SH-SY5Y Cells

ZHANG Lina, LI Mengjie, SHANG Yinghui, LIU Yunru, HUANG Hanchang, LAO Fengxue

(Beijing Key Laboratory of Bioactive Substances and Functional Foods, Institute of Functional Factors and Brain Science, Beijing Union University, Beijing 100191, China)

Online:2022-04-15 Published:2022-04-26

摘要/Abstract

摘要： 目的：探讨玉米黄素对衣霉素（tunicamycin，TM）诱导的SH-SY5Y细胞活性、细胞周期、自噬及凋亡的影响。方法：分别设置空白对照组（不作处理）、玉米黄素组（5 μmol/L玉米黄素）、TM损伤组（5 μg/mL TM）和损伤加保护组（5 μmol/L玉米黄素预处理＋5 μg/mL TM进一步共处理），采用噻唑蓝法检测各处理对SH-SY5Y细胞存活率的影响，从而筛选药物共处理时间；采用流式细胞术测定细胞周期的变化；通过Western blot法测定自噬相关蛋白Beclin1、Beclin1-C和微管结合蛋白1轻链3（microtubule-associated protein1 light chain 3，LC3）B以及凋亡相关蛋白B淋巴细胞瘤-2（B-cell lymphoma 2，BCL2）的表达量。结果：玉米黄素（5 μmol/L）能减轻TM对SH-SY5Y细胞存活率的抑制作用，与TM损伤组相比，损伤加保护组在处理时间为36 h时差异极显著（P＜0.01），48 h时差异显著（P＜0.05）；而TM损伤组相较于空白对照组在处理24、36、48 h时均表现出极显著差异（P＜0.01）。与空白对照组相比，TM处理可以极显著增加细胞周期中G0/G1期细胞数量（P＜0.01），显著或极显著增加LC3B（P＜0.01）、Beclin1（P＜0.05）及Beclin1-C（P＜0.05）表达水平，极显著降低G2/M期细胞数量（P＜0.01）、抗凋亡蛋白BCL2表达水平（P＜0.01）。而玉米黄素联合TM处理与TM损伤组比较，均能减轻TM引起的改变，其中BCL2蛋白水平显著提高（P＜0.05）。结论：玉米黄素可减轻TM处理引起的SH-SY5Y细胞周期G1期阻滞、促细胞自噬与促凋亡作用，其作用机制与玉米黄素对细胞存活率、细胞周期分布和自噬、凋亡相关蛋白表达的调节有关。

关键词: 玉米黄素, SH-SY5Y细胞, 细胞周期, 自噬, 凋亡

Abstract: Objective: To investigate the effect of co-treatment with zeaxanthin (Zea) and tunicamycin (TM) on the viability, cell cycle, autophagy and apoptosis of SH-SY5Y cells. Methods: The cells were divided into four groups: blank control (without drug treatment), Zea (5 μmol/L), TM-induced injury (5 μg/mL) and Zea (5 μmol/L) + TM (5 μg/mL). The methylthiazol tetrazolium (MTT) method was used to evaluate the survival rate of SH-SY5Y cells to determine the drug co-treatment time for further experiments; flow cytometry was used to determine the change of cell cycle; Western blot was used to determine the expression of autophagy-related proteins such as Beclin1, Beclin1-C and microtubule-associated protein 1 light chain 3B (LC3B), and apoptosis-related protein B-cell lymphoma 2 (BCL2). Results: Zea treatment (5 μmol/L) reduced the inhibitory effect of TM on the survival rate of SH-SY5Y cells. Compared to the injury group, a significant difference in the survival rate of SH-SY5Y cells was observed for the combined treatment group at 36 (P < 0.01) and 48 h (P < 0.05). In addition, compared to the blank control group, TM treatment displayed a significant difference in the survival rate of SH-SY5Y cells at 24, 36, and 48 h (P < 0.01). Compared to the blank group, TM treatment significantly increased the number of G0/G1 phase cells in the cell cycle (P < 0.01) and the expression of LC3B (P < 0.01), Beclin1 (P < 0.05) and Beclin1-C (P < 0.05), and significantly reduced the number of G2/M phase cells (P < 0.01) and the expression level of anti-apoptotic protein BCL2 (P < 0.01). The combined treatment alleviated the changes caused by TM, and significantly increased BCL2 protein expression compared to the injury group (P < 0.05). Conclusion: Zea can significantly extenuate TM-induced cell cycle G1 arrest, autophagy and apoptosis of SH-SY5Y cells, and its underlying mechanism is related to the modulatory of Zea on cell survival, cell cycle distribution, and the expression of autophagy and apoptosis-related proteins.

Key words: zeaxanthin, SH-SY5Y cells, cell cycle, autophagy, apoptosis

中图分类号:

TS201.4

章丽娜, 李梦洁, 商迎辉, 刘芸如, 黄汉昌, 劳凤学. 玉米黄素对衣霉素诱导的SH-SY5Y细胞周期、自噬、凋亡的保护作用[J]. 食品科学, 2022, 43(7): 112-119.

ZHANG Lina, LI Mengjie, SHANG Yinghui, LIU Yunru, HUANG Hanchang, LAO Fengxue. Protective Effect of Zeaxanthin on Tunicamycin-Induced Cell Cycle Arrest, Autophagy and Apoptosis of SH-SY5Y Cells[J]. FOOD SCIENCE, 2022, 43(7): 112-119.

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玉米黄素对衣霉素诱导的SH-SY5Y细胞周期、自噬、凋亡的保护作用

Protective Effect of Zeaxanthin on Tunicamycin-Induced Cell Cycle Arrest, Autophagy and Apoptosis of SH-SY5Y Cells

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