基于脂肪变性细胞模型研究明桑降脂茶调控LO2细胞脂代谢的作用

Regulation of Mingsang lipid-lowering tea on lipidmetabolism of LO2 cells based on steatosis cell model

中文关键词:  明桑降脂茶  含药血清  高效液相色谱法  LO2细胞  脂代谢

英文关键词:Mingsang lipid-lowering tea  medicine-containing serum  high performance liquid chromatography  LO2 cells  lipid metabolism

中文摘要:

      目的 明确明桑降脂茶的降脂作用及其发挥降脂作用的物质基础。方法 制备大鼠空白血清、明桑降脂茶全方和桑叶减味含药血清, 以油酸诱导人正常肝细胞(LO2细胞)建立脂肪变性细胞模型, 将细胞分为对照组(含空白血清)、模型组(含空白血清)、明桑降脂茶全方血清处理组、桑叶减味血清处理组。培养4 h后, 显微镜下观察各组脂肪积累情况; 酶标仪测定细胞内脂质含量; 按试剂盒说明测定细胞内甘油三酯(triglyceride, TG)、总胆固醇(total cholesterlo, TC)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol, HDL-C)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol, LDL-C)含量, 以及过氧化物酶体增殖物启动受体α (peroxisome proliferator-activated receptor alpha, PPARα)、固醇调节元件结合蛋白-1c (sterol regulatory element binding protein 1c, SREBP-1c)表达。高效液相色谱法(high performance liquid chromatography, HPLC)研究各组指纹图谱、绿原酸和橙黄决明素含量的差异。结果 与模型组比较, 明桑降脂茶全方血清组、桑叶减味血清组均能显著降低细胞内脂滴数量和脂质含量, 降低TG、TC、LDL-C含量和SREBP-1c表达, 升高HDL-C含量和PPARα表达, 全方血清组的降脂作用优于桑叶减味血清组; 全方血清组、桑叶减味血清组、空白血清组共有峰分别为53、35、23个, 三者共有峰18个; 剔除共有峰后, 全方血清组有22个不同于桑叶减味血清组的差异峰, 桑叶减味血清组有4个不同于全方血清组的差异峰; 全方血清组中绿原酸和橙黄决明素含量均高于桑叶减味血清组, 绿原酸含量在二者之间差异显著。结论 明桑降脂茶具有明显的降脂作用, 桑叶在其中发挥重要降脂作用。

英文摘要:

      Objective To clarify the lipid-lowering effect of Mingsang lipid-lowering tea and the material basis of its lipid-lowering effect. Methods The blank serum of rats, the medicine-containing serum of the whole prescription and reduced Folium mori of Mingsang lipid-lowering tea were prepared. Steatosis cell model of LO2 cells was established by oleic acid induction. The cells were divided into control group (containing blank serum), model group (containing blank serum), medicine-containing serum treatment group of the whole prescription and medicine-containing serum treatment group of reduced Folium mori. After culture for 4 hours, the fat accumulation of each group was observed under the microscope. The intracellular lipid content was measured by enzyme marker. The levels of triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and the expression of peroxisome proliferator-activated receptor-alpha (PPAR α) and sterol regulatory element binding protein-1c (SREBP-1c) were determined according to the instructions of the kits. High performance liquid chromatography (HPLC) was used to study the differences of the fingerprint, and the contents of chlorogenic acid and citrulline in each group. Results Compared with the model group, the numbers of lipid droplets, the content of lipid, TG, TC, LDL-C, and the expression of SREBP-1c were significantly decreased, the content of HDL-C and the expression of PPARα were significantly increased in medicine-containing serum treatment groups of the whole prescription and reduced Folium mori. The lipid-lowering effect of the whole prescription serum group was better than that of reduced Folium mori serum group. There were 53, 35 and 23 peaks in the whole prescription serum, the reduced Folium mori serum and the blank serum respectively, and there were 18 common peaks among them. After eliminating the common peaks, there were 22 different peaks in the whole prescription serum group compared with the reduced Folium mori serum group, and 4 different peaks in the reduced Folium mori serum group compared with the whole prescription serum group. The content of chlorogenic acid and citrullinum in the whole prescription serum group were higher than those in the reduced Folium mori serum group, and the difference of chlorogenic acid content between them was significant. Conclusion Mingsang lipid-lowering tea has obvious lipid-lowering effect, in which Folium mori has played an important role in lipid-lowering.