Caveolin-1结合Cyclophilin A改善大鼠血管平滑肌细胞胆固醇蓄积

Caveolin-1 combined with cyclophilin A improves cholesterol accumulation in rat vascular smooth muscle cells

观察caveolin-1和cyclophilin A在血管平滑肌细胞荷脂过程中的表达变化及其对细胞内胆固醇蓄积的影响。实验采用Western-blot、免疫荧光法分别定量,定位检测caveolin-1和cyclophilin A蛋白的表达;油红O染色观察细胞内脂滴的形成情况;高效液相检测细胞内外总胆固醇含量;免疫共沉淀检测caveolin-1和cyclophilin A相互作用。结果显示,随着荷脂时间延长,caveolin-1和cyclophilin A的蛋白表达逐渐减弱;细胞内脂滴形成逐渐增多,细胞内总胆固醇含量不断增加;caveolin-1和cyclophilin A在大鼠血管平滑肌细胞中相互结合,并且cyclophilin A活性抑制后,与caveolin-1结合能力减低,而细胞内胆固醇含量增加。这提示,caveolin-1和cyclophilin A在血管平滑肌细胞内可相互结合,并参与改善细胞内胆固醇蓄积。

To observe the expression changes of caveolin-1 and cyclophilin A in vascular smooth muscle cells during lipid storage and the effect on intracellular cholesterol accumulation.The protein expression of caveolin-1 and cyclophilin A was detected by Western blotting and immunofluorescence.The formation of lipid droplets in cells was observed by oil red O staining.The total cholesterol content inside and outside the cells was analyzed by HPLC.Co-precipitation was used to detect the interaction between caveolin-1 and cyclophilin A.The results showed that the protein expression of caveolin-1 and cyclophilin A decreased gradually.The formation of intracellular lipid droplets increased and the total cholesterol content in the cells increased continuously.Caveolin-1 and cyclophilin A binds to rat vascular smooth muscle cells,and after inhibition of cyclophilin A activity,the ability to bind to caveolin-1 is decreased,and intracellular cholesterol content is increased.This suggests that caveolin-1 and cyclophilin A bind to each other in vascular smooth muscle cells and are involved in improving intracellular cholesterol accumulation.