Angiotensin Ⅱ Promotes Vascular Smooth Muscle Cells Proliferation and Migration through Inducing Autophagy
Objective To explore the role of autophagy in the process of angiotensin Ⅱ(Ang Ⅱ)induced proliferation and migration of vascular smooth muscle cells(VSMCs).Methods VSMCs from primary cultures of mice were stimulated with Ang Ⅱ(10-7 mol/L)for 24 hours.The expression of autophagy markers LC3B and P62 were detected by immunoblotting.Under the fluorescence microscope,the number of autophagosomes and autolysosomes in the control group(Con group)and Ang Ⅱ group were observed and counted to investigate the effect of Ang Ⅱ on VSMCs autophagy.Changes in VSMCs proliferation and migration after AngⅡ stimulation were evaluated through EdU proliferation assay and scratch assay.After intervention with the autophagy inhibitor 3-MA(10 mmol/L),Western blot and mCherry-GFP-LC3 dual fluorescence indicator system were used to observe and detect changes in LC3 Ⅱ and P62 expression,as well as changes in the number of autophagosomes and autolysosomes,confirming the inhibition of autophagy.Through EdU proliferation experiments and scratch tests,the influence of 3-MA on Ang Ⅱ-induced VSMCs proliferation and migration was examined.Results Compared with the Con group,Ang Ⅱ significantly increased the expression of LC3 Ⅱ and the ratio of LC3 Ⅱ/LC3 Ⅰ,and significantly reduced the expression of P62.Moreover,there was a significant increase in the number of autophagosomes and autolysosomes,indicating that Ang Ⅱ can induce VSMCs autophagy.Ang Ⅱ also significantly increased the number of EdU positive cells and the migration distance of VSMCs,indicating that Ang Ⅱ can promote VSMCs migration and proliferation.Compared with the Ang Ⅱ group,3-MA significantly increases the expression of P62,decreases the expression of LC3Ⅱ and the ratio of LC3 Ⅱ/LC3 Ⅰ,as well as the number of autophagosomes and autolysosomes,inhibiting the occurrence of autophagy.At the same time,3-MA significantly reduces the number of EdU positive cells and the migration distance of VSMCs,inhibiting the proliferation and migration of VSMCs.Conclusion Ang Ⅱ promotes VSMCs proliferation and migration by inducing autophagy.
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