柚皮素增加自噬抑制泡沫细胞的形成

Naringenin inhibits the formation of foam cells by promoting the autophagy

观察柚皮素(naringenin)对THP-1源性泡沫细胞自噬的影响,初步探讨其防治动脉粥样硬化的可能作用机制。首先使用cck8实验检测不同浓度柚皮素对细胞活性的影响。再将不同浓度的柚皮素(0、10、25、50、100μmol/L)作用于泡沫细胞,Western blot检测泡沫细胞中LC3、p62的表达并检测泡沫细胞的总胆固醇含量。然后将柚皮素与雷帕霉素(rapamycin)、3MA分别作用于泡沫细胞后分析LC3、p62的蛋白表达水平。再将柚皮素、compound C分别处理泡沫细胞检测LC3、p-AMPK、AMPK的蛋白表达水平并检测泡沫细胞的总胆固醇(TC)的含量。结果显示,柚皮素对泡沫细胞活性无毒性影响(P>0.05);与模型组相比,柚皮素组LC3II/LC3I比值增加,p62表达减少,总胆固醇含量减少(P<0.05);与模型组相比,柚皮素组LC3II/LC3I比值及p-AMPK/AMPK比值增加,总胆固醇含量减少(P<0.05)。柚皮素能够上调泡沫细胞的自噬能力,减轻细胞内胆固醇的含量从而抑制泡沫细胞的形成;柚皮素通过增加AMPK的磷酸化水平促进泡沫细胞的自噬能力,这可能是柚皮素防治动脉粥样硬化的分子机制之一。

Observe the effect of naringenin on autophagy of THP-1 foam cells to explore its potential mechanism for the prevention and treatment of atherosclerosis.First of all,CCK8 experiment was used to test the effect of naringenin on cell viability at different concentrations.In the next steps,different concentrations of naringenin(0,10,25,50,100μmol/L)were applied to foam cells,and Western blot was used to detect the expression of LC3,p62 and the content of total cholesterol in foam cells.Naringenin,rapamycin and 3MA were applied to foam cells respectively to analyze the protein expression levels of LC3 and p62 later.Ox-LDL,naringenin and compound C were applied to macrophages respectively to analyze the protein expression levels of LC3,p-AMPK,AMPK and the content of total cholesterol finally.The results showed as follows:Naringenin had no toxic effect on foam cells activity(P>0.05);Compared the naringenin group with the model group,the ratio of LC3II/LC3I was increased(P<0.05),and the expression of P62 was reduced(P<0.05),the content of total cholesterol was reduced otherwise(P<0.05);Compared the naringenin group with the model group,the ratio of LC3II/LC3I and the ratio of P-AMPK/AMPK were increased(P<0.05),and the content of total cholesterol was reduced(P<0.05).Naringenin could up-regulate the autophagy ability of foam cells,and reduce the content of total cholesterol in the cells to inhibit the formation of foam cells.Naringenin promoted the autophagy ability of foam cells by increasing the level of AMPK phosphorylation,which might be one of the molecular mechanisms of naringenin in preventing and treating atherosclerosis.