| 摘要: |
| ［摘要］ 目的 构建油酸诱导的HepG2细胞脂肪变性模型，探索水飞蓟宾调节HepG2细胞脂质沉积的作用机制。方法 通过0.4 mM油酸刺激HepG2细胞24 h构建脂肪变性模型。根据实验目的设置对照组、油酸组（OA组）、油酸+水飞蓟宾组（OA+S组）、油酸+氯喹组（OA+CQ组）和油酸+氯喹+水飞蓟宾组（OA+CQ+S组）。每组4样本。通过油红O染色以及甘油三酯(TG)检测评估各干预组细胞脂质沉积情况。通过Western blot检测各干预组自噬标志蛋白LC3B的表达情况。结果 OA组HepG2细胞胞内染色脂滴数量明显多于对照组，且TG含量高于对照组，差异有统计学意义（P<0.05）。OA+S组经水飞蓟宾干预后胞内染色脂滴数量减少，TG含量下降，与OA组比较差异有统计学意义（P<0.05）。OA+CQ+S组在油酸刺激后，用自噬抑制剂氯喹预处理1 h，再加水飞蓟宾干预24 h，HepG2细胞内出现显著脂质沉积，TG含量上升，与OA+S组比较差异有统计学意义（P<0.05），而与OA组水平相当（P>0.05）。Western blot检测结果显示，OA组LC3BⅡ/LC3BⅠ水平较对照组降低（P<0.05）；OA+S组LC3BⅡ/LC3BⅠ水平显著高于OA组，与对照组比较差异无统计学意义（P>0.05）；OA+CQ组、OA+CQ+S组的LC3BⅡ/LC3BⅠ水平显著高于OA组和OA+S组（P<0.05）。结论 0.4 mM油酸可引起HepG2细胞脂肪变性。水飞蓟宾通过增强HepG2细胞自噬而改善油酸引起的脂质沉积。 |
| 关键词: 水飞蓟宾 自噬 油酸 脂肪变性 非酒精性脂肪肝 |
| DOI：10.3969/j.issn.1674-3806.2021.11.07 |
| 分类号:R 575.5 |
| 基金项目:湖南省教育厅科学研究资助项目（编号：17C1152） |
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| Mechanisms of silibinin relieving oleic acid-induced lipid deposition in HepG2 cells |
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XIAO Su-jun, XIAO Ya-jun, WU Pei-sai, et al.
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Teaching and Research Section of Physiology, Hunan University of Medicine, Huaihua 418000, China
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| Abstract: |
| ［Abstract］ Objective To construct a model of fatty degeneration of hepatoblastoma cell line(HepG2) cells induced by oleic acid, and to explore the mechanisms of silibinin in regulating lipid deposition in HepG2 cells. Methods HepG2 cells were stimulated by 0.4 mM oleic acid for 24 hours to construct a steatosis model. The control group, the oleic acid group(OA group), the oleic acid+silibinin group(OA+S group), the oleic acid+chloroquine group(OA+CQ group) and the oleic acid+chloroquine+silibinin group(OA+CQ+S group) were set up according to different experimental purposes, with 4 samples in each group. Oil red O staining and triglyceride(TG) detection were used to evaluate the lipid deposition of cells in each intervention group. Western blot was used to detect the expression of autophagy marker protein LC3B in each intervention group. Results The number of intracellular stained lipid droplets in HepG2 cells in the OA group was significantly more than that in the control group, and the TG content in the OA group was higher than that in the control group, and the differences were statistically significant(P<0.05). In the OA+S group, the number of intracellular stained lipid droplets decreased after the intervention of silibinin, and the content of TG decreased, which were significantly different compared with those in the OA group(P<0.05). The OA+CQ+S group was pre-treated with autophagy inhibitor chloroquine for 1 hour after stimulation with oleic acid, and then added with silibinin for 24 hours. Significant lipid deposition occurred and TG content increased in HepG2 cells, and there were statistically significant differences between the OA+CQ+S group and the OA+S group(P<0.05), but the changes of lipid deposition and the increase of TG content in the OA+CQ+S group were similar to those in the OA group(P>0.05). The results of Western blot detection showed that the level of LC3BⅡ/LC3BⅠ in the OA group was lower than that in the control group(P<0.05), and the level of LC3BⅡ/LC3BⅠin the OA+S group was significantly higher than that in the OA group, and there was no significant difference in the level of LC3BⅡ/LC3BⅠbetween the OA+S group and the control group(P>0.05). The levels of LC3BⅡ/LC3BⅠin the OA+CQ group and the OA+CQ+S group were significantly higher than those in the OA group and the OA+S group(P<0.05). Conclusion 0.4 mM oleic acid can cause fatty degeneration of HepG2 cells. Silibinin improves oleic acid-induced lipid deposition by enhancing autophagy in HepG2 cells. |
| Key words: Silibinin Autophagy Oleic acid Steatosis Nonalcoholic fatty liver disease |
