Page 32 - 《中国药房》2022年11期

洋甘菊总黄酮对HepG2细胞脂质沉积的改善作用研究 Δ

阿曼古丽·艾则孜 1，2＊ ，马红梅 ，兰 卫 （1.新疆医科大学中医学院，乌鲁木齐 830017；2.新疆维吾尔医学专科
1 #
1
学校药学系，新疆 和田 848000）

中图分类号 R965 文献标志码 A 文章编号 1001-0408（2022）11-1306-07
DOI 10.6039/j.issn.1001-0408.2022.11.05

摘 要 目的 探讨洋甘菊总黄酮对人肝癌HepG2细胞脂质异常的调节作用及其降脂机制。方法 采用大孔树脂分离纯化得到高
含量的洋甘菊总黄酮提取物。将HepG2细胞分为对照组（不给药）、模型组（不给药）、非诺贝特组（阳性对照，3.61 μg/mL）和洋甘
菊总黄酮低、中、高剂量组（100、150、200 μg/mL）后，除对照组不造模外，其余组细胞采用1 mmol/L油酸和棕榈酸混合液联合诱导
建立脂质沉积细胞模型。干预24 h后，检测细胞上清液中的游离脂肪酸（FFA）水平和细胞中的三酰甘油（TG）、FFA水平；采用油
红Ｏ染液染色观察细胞中脂滴沉积情况，并检测脂质含量；采用DAPI染液染色观察细胞中二酰基甘油酰基转移酶2（DGAT2）蛋
白表达情况，并检测DGAT2蛋白表达荧光强度；采用Western blot法检测TG合成关键酶乙酰辅酶Ａ羧化酶（ACC）、脂肪酸合成酶
（FAS）、DGAT2的蛋白表达水平。结果 经分离纯化后，洋甘菊总黄酮含量从6.72％提高到56.20％。细胞实验结果显示，与对照组
比较，模型组细胞中的TG、FFA水平和细胞上清液中的FFA水平显著升高，细胞中脂质含量显著升高，DGAT2蛋白表达荧光强度
显著升高，ACC、FAS、DGAT2蛋白表达水平均显著升高（P＜0.01）；细胞中可见大量脂滴堆积。与模型组比较，洋甘菊总黄酮低、
中、高剂量组以上指标水平均显著逆转（P＜0.01）；细胞中脂滴堆积现象明显减少。结论 洋甘菊总黄酮可以抑制脂质沉积模型
HepG2细胞的TG合成，减少细胞脂质积累，防止细胞脂质损伤，其机制可能与其下调ACC/FAS/DGAT2通路表达有关。
关键词 洋甘菊总黄酮；脂质沉积；降脂作用

Study on improvement effect of total flavonoids from Matricaria recutita on lipid deposition in HepG2 cells
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Amanguli·Aizezi ，MA Hongmei ，LAN Wei（1. College of Traditional Chinese Medicine，Xinjiang Medical
1，2
University，Urumqi 830017，China；2. Dept. of Pharmacy，Xinjiang Uygur Medical College，Xinjiang Hotan
848000，China）

ABSTRACT OBJECTIVE To investigate the regulatory effect of total flavonoids of Matricaria recutita on lipid abnormalities in
human hepatoma HepG2 cells and its lipid-lowering mechanism. METHODS The high-content total flavonoids extract from M.
recutita was isolated and purified by macroporous resin. HepG2 cells were divided into control group（without administration），
model group（without administration），fenofibrate group（positive control，3.61 μg/mL）and M. recutita total flavonoids low-dose，
medium-dose and high-dose groups（100，150 and 200 μg/mL）. Except for control group，lipid deposition model of HepG2 cells in
other groups were established by 1 mmol/L mixture of oleic acid and palmitic acid. After 24 hours of intervention，the levels of free
fatty acids（FFA）in cell supernatant and triglyceride（TG）and FFA in cells were detected；Oil red O staining was used to observe
the deposition of lipid droplets in cells and detect the content of lipid；DAPI staining was used to observe the protein expression of
diacylglycerol acyltransferase 2（DGAT2）in cells，and fluorescence intensity of protein expression of DGAT2 were also detected；
protein expressions of key enzymes of TG synthesis as acetyl CoA carboxylase（ACC），fatty acid synthase（FAS）and DGAT2
were detected by Western blot. RESULTS After separation and purification，the content of total flavonoids from M. recutita
increased from 6.72％ to 56.20％. The results of cell experiment showed that compared with control group，the levels of TG and
FFA in cells and FFA in the cell supernatant increased significantly in the model group，the content of lipid in cells increased
significantly，the fluorescence intensity of protein expression of DGAT2 increased significantly，and the protein expressions of
ACC，FAS and DGAT2 increased significantly（P＜0.01）；a
Δ 基金项目：国家自然科学基金-地区科学基金资助项目（No.
81960771）；国家重点研发计划项目（No.2017YFC1703902）；新疆维吾 large number of lipid droplets were accumulated in the cells.
尔自治区自然科学基金资助项目（No.2018D01C160）；新疆维吾尔自 Compared with model group，the levels of above indexes in
治区高校科研计划自然科学青年项目（No.XJEDU2017S025） M. recutita total flavonoids low-dose， medium-dose and
＊讲师，硕士研究生。研究方向：中药药理。E-mail：297475833
high-dose groups were significantly reversed （P＜0.01）；the
@qq.com
accumulation of lipid droplets in cells decreased significantly.
# 通信作者：教授，博士生导师，博士。研究方向：中药药理。电
话：0901-2110289。E-mail：lanwei516@sina.com CONCLUSIONS M. recutita total flavonoids can inhibit the

·1306 · China Pharmacy 2022 Vol. 33 No. 11 中国药房 2022年第33卷第11期