山橿中生物碱类成分Laetanine、Launobine对乙酸致GES-1细胞损伤的保护作用及机制
Protective effect and mechanism of alkaloid components Laetanine and Launobine from Lindera reflexa Hemsl.on GES-1 cell damage induced by acetic acid
李文博 1任伟宏 1孙孝亚2
作者信息
- 1. 河南中医药大学第一附属医院检验科,河南郑州 450000
- 2. 河南中医药大学药学院,河南郑州 450046;河南省中药资源与中药化学重点实验室,河南郑州 450046;河南羚锐制药股份有限公司,河南信阳 465550
- 折叠
摘要
目的 利用乙酸建立胃黏膜上皮细胞(GES-1 细胞)损伤模型,研究山橿中生物碱类成分Laetanine、Launobine对GES-1 细胞损伤的保护作用及机制.方法 通过MTT细胞增殖/毒性实验确定Laetanine、Launobine的最佳给药浓度.分别以浓度为 0.01%～0.2%的乙酸培养液作用于GES-1 细胞,作用时间分别为 3、4、5h,筛选最佳造模条件.利用乙酸建立GES-1 细胞损伤模型,测定Laetanine、Launobine含药培养基处理后的细胞存活率;Griess法测定细胞上清液中NO的浓度;ELISA法测定各组细胞上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和前列腺素E2(PGE2)水平;WST-1 法检测各实验组超氧化物歧化酶(SOD)抑制率,计算SOD活力值.结果 0.1%乙酸溶液处理 3h为GES-1 细胞损伤的最佳造模条件;与模型组比较,Laetanine、Launobine给药组均能显著升高GES-1 细胞存活率(P<0.01);经Laetanine、Launobine处理后,细胞上清液中的NO、TNF-α、IL-6 和PGE2 水平显著降低,SOD活力显著升高(P<0.01 或P<0.05).结论 山橿中生物碱类成分Laetanine和Launobine均能保护乙酸损伤的GES-1 细胞,减轻GES-1 细胞的受损程度,其作用机制可能与其降低NO、TNF-α、IL-6 和PGE2 水平,升高SOD水平有关,表明Laetanine和Launobine可能为山橿发挥抗胃溃疡作用的有效成分.
Abstract
Objective To establish the injury model of human gastric mucosal epithelial cells(GES-1 cells)with acetic acid,and to explore the protective effect and mechanism of the alkaloid components Laetanine and Launobine from Lindera reflexa Hemsl.on the injury of GES-1 cells.Methods The optimal concentration of Laetanine and Launobine were determined by MTT cell proliferation/toxicity experiment.The culture medium containing 0.01%～0.2%acetic acid was used to act on GES-1 cells for 3,4 and 5 hours,respectively.The best modeling conditions were screened.The cell survival rate of GES-1 cells treated with Laetanine and Launobine was determined by acetic acid-induced cell injury model.The concentration of NO in cell supernatant was measured by Griess method.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and prostaglandin E2(PGE2)secreted in cell supernatant were measured by ELISA.The inhibitory rate of superoxide dismutase(SOD)was measured by WST-1,and the activity value of SOD was calculated.Results The treatment with 0.1%acetic acid solution for 3h was the best modeling condition for GES-1 cells injury.Compared with the model group,both Laetanine and Launobine could significantly increase the survival rate of GES-1 cells(P<0.01).After treatment with Laetanine and Launobine,the levels of NO,TNF-α,IL-6 and PGE2 in cell supernatant were significantly decreased,while the activity of SOD was significantly increased(P<0.01 or P<0.05).Conclusion The alkaloid components Laetanine and Launobine from Lindera reflexa Hemsl.can both protect GES-1 cells injured by acetic acid,reduce the degree of cells damage.The mechanism of action may be related to the decrease of NO,TNF-α,IL-6 and PGE2 levels and the increase of SOD level.These results indicate that Laetanine and Launobine may be effective components of Lindera reflexa Hemsl.for anti-gastric ulcer.
关键词
山橿/生物碱/Laetanine/Launobine/胃溃疡/GES-1细胞/保护/机制Key words
Lindera reflexa Hemsl./Alkaloid/Laetanine/Launobine/Gastric ulcer/GES-1 cells/Protection/Mechanism引用本文复制引用
基金项目
河南省科技攻关计划(212102310353)
出版年
2024
NETL
NSTL
万方数据