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目的:观察化瘀祛痰方含药血清对油酸和棕榈酸诱导HepG2细胞脂质损伤及自噬水平的影响,探讨化瘀祛痰方防治脂代谢紊乱的可能机制。方法:以不同终浓度(摩尔比为2∶1)的油酸和棕榈酸混合物孵育诱导HepG2细胞24h,采用CCK-8、油红O染色以及细胞内TG含量测定等方法筛选并建立最佳HepG2细胞脂质损伤模型;以大鼠空白血清作为空白对照、辛伐他汀含药血清作为阳性对照,采用油红O染色、Western Blot、细胞内TG含量测定法检测化瘀祛痰方含药血清对HepG2细胞脂肪损伤及自噬相关指标的影响。结果:选取终浓度为1mmol/L的油酸和棕榈酸作为最佳作用浓度。与模型组比较,化瘀祛痰方含药血清能明显使HepG2细胞内脂滴变小、数量减少并使细胞内TG含量显著减低(P<0.05),同时显著上调HepG2细胞自噬相关蛋白Atg3和Beclin1表达(P<0.05)。结论:化瘀祛痰方药含药血清可以减轻油酸和棕榈酸诱导的HepG2细胞内的脂质损伤,其机制可能与其影响自噬相关蛋白Atg3和Beclin1的表达有关。
OBJECTIVE: To observe the effect of medicated serum of Huayu Qutan Recipe on lipid damage and autophagy of HepG2 cells induced by oleic acid and palmitic acid, and to explore the possible mechanism of preventing blood lipid disorder by treating Huayu Qutan recipe. Methods: HepG2 cells were induced by incubation with a mixture of oleic acid and palmitic acid at different final concentrations (molar ratio of 2: 1) for 24 h. The optimal HepG2 cells were screened by CCK-8, oil red O staining and intracellular TG assay The model of lipid-induced cell injury was established. The blank serum of rats was used as a blank control and the simvastatin-containing serum was used as a positive control. Oil red O staining, Western Blot and intracellular TG assay were used to detect the drug-containing serum of Huayu Qutan Recipe Effects of HepG2 cells on fat injury and autophagy related indexes. Results: Oleic acid and palmitic acid with the final concentration of 1mmol / L were selected as the best concentration. Compared with the model group, the drug-containing serum of Huayu Qutan Recipe could obviously decrease the number of lipid droplets and decrease the content of TG in HepG2 cells (P <0.05), and significantly up-regulate the expression of Atg3 And Beclin1 expression (P <0.05). Conclusion: Medicated serum of Huayu Qutan Recipe can reduce the lipid damage in HepG2 cells induced by oleic acid and palmitate, which may be related to its effect on the expression of autophagy-related proteins Atg3 and Beclin1.