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投稿时间：2021-08-26
投稿时间：2021-08-26
中文摘要: 为研究苦荞总黄酮（total flavonoids of tartary buckwheat，TFTB）对体外诱导的酒精性脂肪肝（alcoholic fatty liver，AFL）L-02细胞的影响。采用细胞增殖-毒性检测试剂盒（cell counting kit-8，CCK-8）筛选AFL细胞模型乙醇诱导浓度、TFTB作用浓度，油红O染色观察AFL细胞模型构建情况，用试剂盒方法检测TFTB作用前后细胞上清液中甘油三酯（triglyceride，TG）、天冬氨酸氨基转移酶（aspartate transaminase，AST）、丙氨酸氨基转移酶（alanine aminotransferase，ALT）、丙二醛（malondialdehyde，MDA）、游离脂肪酸（nonestesterified fatty acid，NEFA）、总超氧化物歧化酶（total superoxide dismutase，T-SOD）和谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）水平，酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测 TFTB 作用前后细胞上清液白介素-6（interleukin-6，IL-6）、白介素-8（interleukin-8，IL-8）和肿瘤坏死因子（tumor necrosis factor-α，TNF-α）的水平变化。用含 0.9%乙醇和 0.01%FeSO4的RPMI-1640培养液诱导L-02细胞，诱导至第4代，与空白对照组及阴性对照组相比，模型组经油红O染色后胞浆内出现大量橘红色脂滴且产生明显的TG、AST、ALT堆积。与模型组相比，80 μg/mL TFTB处理组细胞上清液中的TG、AST、ALT、MDA、NEFA水平分别降低 15.00%、61.50%、68.71%、13.83%和47.51%，而 T-SOD、GSH-Px水平分别升高17.44%、101.57%。此外，TFTB处理后体外诱导的AFL细胞上清液中IL-6、IL-8和TNF-α水平分别降低8.31%、32.61%、27.85%，表明TFTB不仅可以抑制肝细胞脂肪积累与变性、减轻乙醇对肝细胞膜及肝细胞的氧化损伤，还可以缓解由于AFL细胞损伤所引起的炎症反应。
Abstract:This study aimed to investigate the effect of total flavonoids of tartary buckwheat（TFTB）on the L-02 cells of alcoholic fatty liver（AFL）.To be specific，cell counting kit-8（CCK-8）assay was employed to screen the optimal concentration of ethanol for the induction of AFL cell model and the optimal concentration of TFTB to be tested，and oil red O staining to test the modeling outcome.Levels of triglyceride（TG），aspartate transaminase（AST），alanine aminotransferase（ALT），malondialdehyde（MDA），nonestesterified fatty acid（NEFA），total superoxide dismutase（T-SOD），and glutathione peroxidase（GSH-Px）were determined with corresponding kits and levels of interleukin-6（IL-6），interleukin-8（IL-8），and tumor necrosis factor-α（TNF-α）were detected by enzyme-linked immunosorbent assay（ELISA）.When L-02 cells were cultured with the RPMI-1640 medium containing 0.9%ethanol and 0.01%FeSO4to the fourth generation，a large number of orange lipid droplets were observed in the cytoplasm after oil red O staining and significant accumulation of TG，AST and ALT was detected in the model group as compared with the blank control group and the negative control group.Compared with model group，80 μg/mL TFTB decreased the levels of TG，AST，ALT，MDA and NEFA by 15.00%，61.50%，68.71%，13.83%and 47.51%，respectively，and raised the levels of T-SOD and GSH-Px by 17.44%and 101.57%，separately.In addition，the content of IL-6，IL-8 and TNF-α in the supernatant of AFL cells was respectively 8.31%，32.61%and 27.85%lower after TFTB treatment.All these indicate that TFTB not only inhibits the lipid accumulation and steatosis of liver cells and alleviates the oxidative damage of ethanol to liver cell membranes and liver cells but also relieves the inflammatory response caused by AFL cell injury.
keywords: total flavonoids of tartary buckwheat alcoholic fatty liver cell model steatosis inflammatory factors
文章编号：202223008 中图分类号： 文献标志码：
基金项目:泸州品创科技有限公司横向项目（18H1314）；企业大创项目（C2021115405、202110611594X）；四川省原子能研究院辐照保藏四川省重点实验室开放基金资助（FZBC2020001）
| 作者 | 单位 |
| 刘怡1，张娜1，吴萌萌1，严馨1，李姝1，2，王松涛2，3，沈才洪2，3，黄敏4，赵建1* | 1.四川大学生命科学学院，四川 成都 610064；2.泸州品创科技有限公司，四川 泸州 646000；3.国家固态酿造工程技术研究中心，四川 泸州 646000；4.四川省原子能研究院，四川 成都 610101 |
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