[1]王德宇,杨东亮,常 刘.异鼠李素对H2O2诱导的肠上皮细胞氧化应激损伤的保护作用[J].陕西中医,2023,(1):7-11.[doi:DOI:10.3969/j.issn.1000-7369.2023.01.002]
　WANG Deyu,YANG Dongliang,CHANG Liu.Protective effect of isorhamnetin on H2O2-induced oxidative stress injury of intestinal epithelial cells[J].,2023,(1):7-11.[doi:DOI:10.3969/j.issn.1000-7369.2023.01.002]
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异鼠李素对H2O2诱导的肠上皮细胞氧化应激损伤的保护作用
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2023年1期
页码:
7-11
栏目:
基础研究
出版日期:
2023-01-05

文章信息/Info

Title:
Protective effect of isorhamnetin on H2O2-induced oxidative stress injury of intestinal epithelial cells
作者:
王德宇; 杨东亮; 常 刘
(大连市第四人民医院烧伤创面科,辽宁 大连 116000)
Author(s):
WANG Deyu; YANG Dongliang; CHANG Liu
(Department of Burn Wound,Dalian the Fourth People's Hospital,Dalian 116000,China)
关键词:
异鼠李素;  肠上皮细胞;  H2O2;  氧化应激;  Nrf2/HO-1;  PI3K/Akt
Keywords:
Isorhamnetin;  Enterocyte;  H2O2 ;  Oxidative stress;  Nrf2/HO-1;  PI3K/Akt
分类号:
R 969
DOI:
DOI:10.3969/j.issn.1000-7369.2023.01.002
文献标志码:
A
摘要:
目的:评价异鼠李素对H2O2诱导的大鼠肠上皮细胞IEC-6氧化应激损伤的保护作用及机制。方法:MTT实验筛选异鼠李素对IEC-6细胞的安全作用浓度用于评价抗H2O2诱导的氧化应激损伤功效。设正常对照组、H2O2组、N-乙酰半脱氨酸(NAC)组(10 μmol/L)、异鼠李素低剂量组(20 μmol/L)、异鼠李素中剂量组(40 μmol/L)及异鼠李素高剂量组(100 μmol/L)。正常对照组常规培养,H2O2组用500 μmol/L H2O2处理,NAC及异鼠李素组分别用10 μmol/L NAC,20、40、100 μmol/L异鼠李素预处理细胞相应时间后,用500 μmol/L H2O2继续培养。流式细胞术检测细胞凋亡及ROS水平,试剂盒检测细胞中MDA含量及SOD活性,Western blot检测细胞中Nrf2/HO-1、PI3K/Akt信号通路相关蛋白的表达。结果:与H2O2组相比,异鼠李素高剂量组IEC-6细胞活性显著增强(P<0.05)。流式细胞仪检测发现,异鼠李素高剂量组细胞凋亡率及ROS水平显著降低(P<0.05)。异鼠李素高剂量组细胞中MDA含量降低, SOD活性增强(P<0.05)。Western blot结果显示,异鼠李素高剂量组可增加p-Akt水平、促进Nrf2核移位及HO-1的表达(P<0.05)。结论:异鼠李素可能通过激活Nrf2/HO-1通路,缓解H2O2引起的氧化应激损伤。
Abstract:
Objective:To evaluate the protective effect and mechanism of isorhamnetin on H2O2-induced oxidative damage in IEC-6 cells.Methods:MTT assay was used to screen the safe concentration of isorhamnetin on IEC-6 cells to evaluate the efficacy against H2O2oxidative stress injury.The normal control group,H2O2group,NAC group(10 μmol/L)and isorhamnetin low dose group(20 μmol/L),medium dose group(40 μmol/L)and high dose groups(100 μmol/L)were set.The H2O2 group was routinely cultured,and the model group was treated with 500 μmol/L H2O2.The NAC and isorhamnetin groups were pretreated with 10 μmol/L NAC,20,40,and 100 μmol/L isorhamnetin for corresponding time,and then continued to culture with 500 μmol/L H2O2.Cell apoptosis and ROS levels were detected by flow cytometry,MDA content and SOD activity in cells were detected by kits,and expressions of Nrf2/HO-1 and PI3K/Akt signaling pathway-related proteins in cells were detected by Western blot.Results:Compared with the H2O2group,100 μmol/L isorhamnetin pretreatment could significantly enhance the viability of IEC-6 cells(P<0.05).Flow cytometry showed that pretreatment with high dose isorhamnetin could significantly reduce the apoptosis rate and ROS level(P<0.05).The test results of the kit showed that high dose isorhamnetin pretreatment could reduce the content of MDA and enhance the activity of SOD(P<0.05).Western blot results showed that high dose isorhamnetin pretreatment could increase the level of p-Akt,promote the nuclear translocation of Nrf2 and the expression of HO-1(P<0.05).Conclusion:Isorhamnetin may protect against H2O2-induced oxidative stress injury by activating the Nrf2/HO-1 pathway.

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备注/Memo

备注/Memo:
基金项目:辽宁省大连市财政局 “登峰计划”资助项目[大财指社(2022)06号]
更新日期/Last Update: 2022-12-28