姜黄素对过氧化氢诱导的罗非鱼肝细胞热休克蛋白基因表达的影响

李林明; 张子平; 王若璇; 黄一帆

doi:10.19303/j.issn.1008-0384.2023.09.004

姜黄素对过氧化氢诱导的罗非鱼肝细胞热休克蛋白基因表达的影响

doi: 10.19303/j.issn.1008-0384.2023.09.004

李林明^{1, 2,},
张子平^{1, 2, ,},
王若璇^{1, 2},
黄一帆³

1.
福建农林大学海洋研究院福建省海洋生物技术重点实验室，福建　福州　350002
2.
福建农林大学海洋学院，福建　福州　350002
3.
福建农林大学福建省兽医中药与动物保健重点实验室，福建　福州　350002

基金项目: 福建省高校领军人才项目（660160010）；福建省自然科学基金项目（2023J01488）

详细信息

作者简介:
李林明（1985 —），男，博士，实验师，主要从事水产动物药理学相关研究，E-mail：444853510@qq.com

通讯作者:
张子平（1964 —），男，博士，教授，主要从事水产生物技术相关研究，E-mail： zhangziping@fafu.edu.cn

中图分类号: S917.4
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- 被引次数: 0
出版历程
- 收稿日期: 2023-02-13
- 修回日期: 2023-06-21
- 网络出版日期: 2023-10-25
- 刊出日期: 2023-09-28

Effect of Curcumin on HSP Expression in Tilapia Hepatocytes under Hydrogen Peroxide Stress

LI Linming^{1, 2
,},
ZHANG Ziping^{1, 2
, ,},
WANG Ruoxuan^{1, 2},
HUANG Yifan³

1.
Key Laboratory of Marine Biotechnology of Fujian Province, Institute of Oceanology, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
College of Marine Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
3.
Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China

摘要

摘要: 目的探究热休克蛋白基因在氧化应激和姜黄素作用下的表达差异，了解姜黄素药效的发挥与热休克蛋白基因表达变化的关系，进一步探讨热休克蛋白对氧化应激条件下鱼类肝细胞中的功能，为鱼类氧化应激的毒理学机制及姜黄素的药理学机制提供理论资料。方法对24 h姜黄素预处理（保护组）与无姜黄素预处理（对照组）的罗非鱼肝细胞分别进行0、1、2.5 h 的H₂O₂应激，并对肝细胞5种高分子热休克蛋白基因（HSP70、HSP90α、HSP90β、HSP60、HSP75）和5种小分子热休克蛋白基因（HSP30、HSPB1、 HSPB7、HSPB8、HSPB11）的表达水平进行qPCR检测。结果与不经H₂O₂处理（0 h）相比，H₂O₂作用 1 h能显著提升HSPB1表达量，降低HSP30表达量；H₂O₂作用2.5 h 能显著提升HSP90a、HSPB1的表达量。姜黄素预处理24 h能显著提升HSP70、HSP90α、HSP30、HSPB1的表达量；姜黄素预处理24 h后再进行H₂O₂应激1 h时，HSP70、HSP90α、HSP30的表达量显著高于H₂O₂单独应激（对照组）；姜黄素预处理24 h后再进行H₂O₂应激2.5 h时，HSP90α、HSP30、HSPB1的表达量显著高于H₂O₂单独应激。其余热休克蛋白在试验过程中无显著差异。结论 H₂O₂单独应激可引起罗非鱼肝细胞HSP90a、HSP30、HSPB1基因的表达量变化，而姜黄素预处理能提高多种热休克蛋白基因的表达，提高了罗非鱼肝细胞的抗氧化能力，最终抵御H₂O₂引起的氧化应激并维持细胞活力。
- 罗非鱼 /
- 姜黄素 /
- 热休克蛋白 /
- 氧化应激
Abstract: Objective Expression of heat shock protein (HSP) gene under oxidative stress in the presence of curcumin was studied to understand the toxicological function of the protein and the pharmacological mechanism of curcumin in hepatocytes of Oreochromis nilotica. Methods Tilapia hepatocytes and those pretreated with curcumin for 24 h were subjected to H₂O₂ stress for 0, 1, or 2.5 h. The expressions of HSP70, HSP90α, HSP90β, HSP60, and HSP75, as well as 5 small molecule HSP genes, i.e., HSP30, HSPB1, HSPB7, HSPB8, and HSPB11, in the cells were determined by qPCR. Results The H₂O₂ treatment for 1 h significantly increased the expression of HSPB1 and decreased that of HSP30 in the tilapia hepatocytes, while the treatment that lasted for 2.5 h significantly increased the expressions of HSP90a and HSPB1. A pretreatment of curcumin for 24 h on the cells not only significantly elevated the expressions of HSP70, HSP90α, HSP30, and HSPB1 over control but also on those of HSP70, HSP90α, and HSP30 under 1 h H₂O₂ stress, as well as those on HSP90α, HSP30, and HSPB1 under 2.5 h H₂O₂ stress. However, no significant differences in other HSPs were observed. Conclusion As a spontaneous response of tilapia hepatocytes to oxidative stress, H₂O₂ affected significantly the expressions of HSP90a, HSP30, and HSPB1 in the cells. By pretreating the hepatocytes with curcumin, expressions of these genes could be uplifted boosting the cellular antioxidant capacity to better resist the stress and maintain viability.
- Tilapia /
- curcumin /
- heat shock proteins /
- oxidative stress

HTML全文

图 1 姜黄素预处理对H₂O₂胁迫下罗非鱼肝细胞高分子HSP基因表达的影响

*表示同一处理时间保护组与对照组间差异显著（P ＜ 0.05），不同字母小写字母表示同一处理组中不同时间点之间相比差异显著（P ＜ 0.05）。图2同。

Figure 1. Effect of curcumin pretreatment on HSP expressions of tilapia hepatocytes under H₂O₂ stress

* indicates significant difference from control at relevant time point at P＜0.05; data with different lowercase letters indicate significant differences at different time points in same group at P＜0.05. Same for Fig. 2.

下载: 全尺寸图片幻灯片

图 2 姜黄素与H₂O₂处理对罗非鱼肝细胞sHSP基因表达的影响

Figure 2. Effect of curcumin pretreatment on sHSP expressions of tilapia hepatocytes under H₂O₂ stress

下载: 全尺寸图片幻灯片

表 1 qPCR检测所用的各基因引物

Table 1. Primers used for qPCR

基因 Gene	长度 Length/bp	引物序列（5′–3′） Sequence (5′–3′)
HSP70	107	HSP70F: CATCGCCTACGGTCTGGACAA
HSP70	107	HSP70R: TGCCGTCTTCAATGGTCAGGAT
HSP90α	128	HSP90αF: ATTGCTCAGCTGATGTCCCT
HSP90α	128	HSP90αR: GTGGGATCCGTCAAGCTTTC
HSP90β	117	HSP90βF: AGAACCTCAAGCTGGGTGTG
HSP90β	117	HSP90βR: TCACGTACTCCGAGAGGGAG
HSP60	115	HSP60F: GCGGCTAGCAGTAAGCCTTT
HSP60	115	HSP60R: CGTAGGCTCGTGTGAGGTG
HSP75	103	HSP75F: GTTGGGCCCACCATTCATCC
HSP75	103	HSP75R: TTCTCAACCATCCACGCCAT
HSP30	111	HSP30F: GACTCTGGACCTCCTCCACT
HSP30	111	HSP30R: TGAAACACTCTTCAGGTGTCTGT
HSPB1	106	HSPB1F: CTTTGACCAGACCTTCGGCA
HSPB1	106	HSPB1R: TAGTTCTGGGGCCAGGATGG
HSPB7	119	HSPB7F: ACTTTCACCCACAAGTGCCA
HSPB7	119	HSPB7R: AGCTCATGCTTGGCTGGATT
HSPB8	104	HSPB8F: TGGAAAGTCTGCGTGAACGTC
HSPB8	104	HSPB8R: TCCTGTTTCTCTTCGTGCTTTC
HSPB11	100	HSPB11F: CAGCTTGTTTGGCGATGACC
HSPB11	100	HSPB11R: CCGGTTGAAGAAGTCTCGCT
β-Actin	129	β-Actin-F: TGCGGGATATCATTTGCCTGA
β-Actin	129	β-Actin-R: GAATCCGGCCTTGCACATAC

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