中国临床药理学杂志

红芪多糖通过PERK-Nrf2信号通路改善脂肪变性肝细胞铁死亡的研究

PDF下载

丁海军 ¹ , 李钦 ² , 张磊 ³ , 李芳 ²

中国临床药理学杂志 | 临床与基础桥接研究 2025,41(22): 3214-3220

收起

中国临床药理学杂志 |临床与基础桥接研究 2025 , 41 (22) : 3214 -3220

红芪多糖通过PERK-Nrf2信号通路改善脂肪变性肝细胞铁死亡的研究

全屏

丁海军¹, 李钦², 张磊³, 李芳²

作者信息

^1.甘肃卫生职业学院药学院，甘肃兰州 730000

^2.湖南医药学院药学院，湖南怀化 418000

^3.甘肃中医药大学基础医学院，甘肃兰州 730000

通讯作者:

李芳，副教授 MP: 13919959028 E-mail: 119232388@qq.com

作者简介:

丁海军（1974-），副教授，主要从事中药药理作用机制研究方面的研究

Research on the improvement of ferroptosis in hepatic steatosis cells by hedysarum polybotrys polysacchcaide through the PERK-Nrf2 signaling pathway

Hai-jun DING¹, Qin LI², Lei ZHANG³, Fang LI²

Affiliations

^1.School of Pharmacy, Gansu Health Vocational College, Lanzhou 730000, Gansu Province, China

^2.School of Pharmaceutical Sciences, Hunan University of Medicine, Huaihua 418000, Hunan Province, China

^3.School of Basic Medicine, Gansu University of Chinese Medicine, Lanzhou 730000, Gansu Province, China

出版时间: 2025-11-28 doi: 10.13699/j.cnki.1001-6821.2025.22.010

文章导航

摘要

收起

目的

探讨红芪多糖（HPS）通过蛋白激酶RNA样内质网激酶-对稳定抗氧化调节蛋白核因子E2相关因子2（PERK-Nrf2）信号通路改善脂肪变性肝细胞铁死亡的机制研究。

方法

用0.6 mmol·L^-1的脂肪酸进行体外诱导培养LO-2细胞，构建非酒精性脂肪性肝病细胞模型。将细胞分为正常组（细胞正常培养）、模型组（0.6 mmol·L^-1脂肪酸干预12 h造模）、阳性对照组（造模后用24 μmol·L^-1 liproxstain-1处理）和实验组（造模后用50 mg·L^-1 HPS处理），各组细胞均培养24 h。用油红O染色法分析细胞脂质沉积阳性面积（%Area）；用酶联反应（ELISA）检测细胞脂质过氧化物（LPO）、谷胱甘肽（GSH）、4-羟基壬烯酸（4-HNE）和超氧化物歧化酶（SOD）的含量；用蛋白质印迹（WB）法和反转录聚合酶链反应（RT-PCR）检测肝细胞PERK-Nrf2信号通路相关蛋白和Messenger RNA（mRNA）的相对表达水平；用免疫荧光法检测谷胱甘肽过氧化酶4（GPX4）及溶质载体家族7成员11（SLC7A11）蛋白相对表达水平。

结果

正常组、模型组、对照组和实验组肝细胞脂质沉积阳性面积分别为0.02±0.01、0.26±0.08、0.09±0.02和0.15±0.04；LPO含量分别为（2.62±0.17）、（13.47±1.55）、（6.31±1.12）和（5.71±1.50）nmol·mL^-1；4-HNE含量分别为（8.06±0.40）、（14.60±2.02）、（9.40±0.81）和（12.94±0.56）ng·L^-1；GSH含量分别为（120.73±7.75）、（34.60±2.98）、（67.86±2.08）和（59.33±7.46）mg·L^-1；SOD含量分别为（7.25±0.61）、（3.77±0.44）、（5.22±0.76）和（8.12±0.78）U·ml^-1；葡萄糖调节蛋白78（GRP78）mRNA相对表达水平分别为1.01±0.15、1.47±0.11、1.41±0.08和1.02±0.05；蛋白激酶RNA样内质网激酶（PERK）mRNA相对表达水平分别为1.00±0.09、1.85±0.16、1.02±0.27和1.16±0.01；核因子E2相关因子2（Nrf2）mRNA表达水平分别为1.01±0.12、0.26±0.07、0.68±0.09和0.66±0.05；GPX4 mRNA相对表达水平分别为1.00±0.05、0.79±0.02、0.87±0.17和0.83±0.10；SLC7A11 mRNA相对表达水平分别为1.01±0.17、0.50±0.14、0.91±0.09和0.98±0.05；Grp78蛋白相对表达水平分别为1.00±0.11、2.72±0.07、1.83±0.04和2.24±0.06；PERK蛋白相对表达水平分别为1.00±0.06、1.21±0.08、1.03±0.07和0.95±0.06；磷酸化（p-）PERK蛋白相对表达水平分别为1.00±0.04、2.70±0.08、1.40±0.07和1.76±0.06；PERK蛋白磷酸化水平分别为1.00±0.09、2.24±0.21、1.36±0.03和1.85±0.09；Nrf2蛋白相对表达水平分别为1.00±0.03、0.52±0.01、0.73±0.02和0.84±0.02；GPX4蛋白荧光强度分别为24.34±0.86、13.86±1.65、18.01±2.03和16.20±1.55；SLC7A11蛋白荧光强度分别为69.58±4.13、34.24±6.73、60.75±9.35和45.18±6.46。模型组的上述指标与正常组比较、实验组的上述指标与模型组比较，在统计学上差异均有统计学意义（均P<0.05）。

结论

HPS可能通过调控PERK-Nrf2信号通路，减轻肝细胞脂质过氧化，减轻铁死亡，从而保护肝细胞。

关键词

红芪多糖 / 非酒精性脂肪肝病 / 蛋白激酶RNA样内质网激酶-对稳定抗氧化调节蛋白核因子E2相关因子2信号通路 / 脂质过氧化 / 铁死亡

Abstract

收起

Objective

To explore the mechanism by which hedysarum polybotrys polysaccharide (HPS) improves iron death of steatotic hepatocytes through the protein kinase RNA-like endoplasmic reticulum kinase- nuclear factor-erythroid 2-related factor 2 (PERK-Nrf2) signaling pathway.

Methods

The cells were cultured with 0.6 mmol·L^-1 fatty acids to replicate the non-alcoholic fatty liver disease cell model. The LO-2 cell were divided into normal group (complete medium), model group (0.6 mmol·L^-1 fatty acid solution), positive control group (0.6 mmol·L^-1 fatty acid solution+24 μmol·L^-1 liproxstain-1) and experimental group (0.6 mmol·L^-1 fatty acid solution + 50 mg·L^-1 HPS), culture for 24 h. Stain with Oil Red O was used to analyze the positive area (%Area) of cell lipid deposition; the content of lipid peroxide(LPO), 4-hydroxynonenal(4-HNE), glutathione(GSH) and superoxide dismutase(SOD) was detected by enzyme-linked immunosorbent assay(Elisa) method; the relative expressions levels of PERK-Nrf2 signaling pathway protein in hepatocytes were detected by Western blot; immunofluorescence assay was used to detect the relative expression levels of GPX4 and SLC7A11 proteins.

Results

The % Area of cell lipid deposition in hepatocytes of normal group, model group, control group and experimental group were 0.02±0.01, 0.26±0.08, 0.09±0.02 and 0.15±0.04; the contents of LPO were (2.62±0.17), (13.47±1.55), (6.31±1.12) and (5.71±1.50) nmol·mL^-1; the contents of 4-HNE were (8.06±0.40), (14.60±2.02), (9.40±0.81) and (12.94±0.56) ng·L^-1; the contents of GSH were (120.73±7.75), (34.60±2.98), (67.86±2.08) and (59.33±7.46) mg·L^-1; the contents of SOD were (7.25±0.61), (3.77±0.44), (5.22±0.76) and (8.12±0.78) U·mL^-1; the levels of Grp78 mRNA were 1.01±0.15, 1.47±0.11, 1.41±0.08 and 1.02±0.05; the levels of PERK mRNA were 1.00±0.09, 1.85±0.16, 1.02±0.27 and 1.16±0.01; the levels of Nrf2 mRNA were 1.01±0.12, 0.26±0.07, 0.68±0.09 and 0.66±0.05; the levels of GPX4 mRNA were 1.00±0.05, 0.79±0.02, 0.87±0.17 and 0.83±0.10; the levels of SLC7A11 mRNA were 1.01±0.17, 0.50±0.14, 0.91±0.09 and 0.98±0.05; the relative expression levels of Grp78 protein were 1.00±0.11, 2.72±0.07, 1.83±0.04 and 2.24±0.06; the relative expression levels of PERK protein were 1.00±0.06, 1.21±0.08, 1.03±0.07 and 0.95±0.06; the relative expression levels of p-PERK protein were 1.00±0.04, 2.70±0.08, 1.40±0.07 and 1.76±0.06; the relative levels of PERK protein phosphorylation were 1.00±0.09, 2.24±0.21, 1.36±0.03 and 1.85±0.09; the relative expression levels of Nrf2 protein were 1.00±0.03, 0.52±0.01, 0.73±0.02 and 0.84±0.02; the protein fluorescence intensity of GPX4 were 24.34±0.86, 13.86±1.65, 18.01±2.03 and 16.20±1.55; the protein fluorescence intensity of SLC7A11 were 69.58±4.13, 34.24±6.73, 60.75±9.35 and 45.18±6.46. Compared with the normal group, there were significant differences in the above indexes of model group (all P<0.05) ; compared with the model group, there were significant differences in the above indexes of experimental group (all P<0.05).

Conclusion

HPS may protect liver cells by regulating the PERK-Nrf2 signaling pathway, reducing lipid peroxidation and iron death in liver cells.

Key words

hedysarum polybotrys polysacchcaide / non-alcoholic fatty liver disease / protein kinase RNA-like endoplasmic reticulum kinase- nuclear factor-erythroid 2-related factor 2 signaling pathway / lipid peroxidation / ferroptosis

引用本文

丁海军, 李钦, 张磊, 李芳. 红芪多糖通过PERK-Nrf2信号通路改善脂肪变性肝细胞铁死亡的研究. 中国临床药理学杂志, 2025 , 41 (22) : 3214 -3220 . DOI: 10.13699/j.cnki.1001-6821.2025.22.010

Hai-jun DING, Qin LI, Lei ZHANG, Fang LI. Research on the improvement of ferroptosis in hepatic steatosis cells by hedysarum polybotrys polysacchcaide through the PERK-Nrf2 signaling pathway[J]. Chinese Journal of Clinical Pharmacology, 2025 , 41 (22) : 3214 -3220 . DOI: 10.13699/j.cnki.1001-6821.2025.22.010

基金

收起

2024年甘肃省高校教师创新基金资助项目(2024A-317)
2023年度国家自然科学基金地区基金资助项目(82360914)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

2025年第41卷第22期

PDF下载

110

引用本文

BibTeX

文章信息

doi: 10.13699/j.cnki.1001-6821.2025.22.010

接收时间：2025-05-12
首发时间：2026-08-05
出版时间：2025-11-28

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2025-05-12

基金

2024年甘肃省高校教师创新基金资助项目(2024A-317)

2023年度国家自然科学基金地区基金资助项目(82360914)

作者信息

^1.甘肃卫生职业学院药学院，甘肃兰州 730000

^2.湖南医药学院药学院，湖南怀化 418000

^3.甘肃中医药大学基础医学院，甘肃兰州 730000

通讯作者:

李芳，副教授 MP: 13919959028 E-mail: 119232388@qq.com

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zglcylxzz/CN/10.13699/j.cnki.1001-6821.2025.22.010

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT