Effect of Reserpine on the autophagy of rat aortic vascular smooth muscle cells
WANG Shunmin1 YANG Xiaodan2 ZENG Yong3 HUANG Lu1 YONG Su′nan1▲
1.Department of Cardiology, the First Hospital of Hu′nan University of Chinese Medicine, Hu′nan Province, Changsha 410007, China;
2.Department of Cardiology, the First Clinical College of Traditional Chinese Medicine, Hu′nan University of Chinese Medicine, Hu′nan Province, Changsha 410208, China;
3.Department of Emergency, the First Hospital of Hu′nan University of Chinese Medicine, Hu′nan Province, Changsha 410007, China
Abstract：Objective To explore the relationship of Reserpine and the autophagy of rat aortic vascular smooth muscle cells (RA-VSMC). Methods RA-VSMC was cultured in medium with Reserpine or Earles balanced salt solution (EBSS) in vitro. MTT assay was used to detect the effect of Reserpine on the proliferation of RA-VSMC with 5 μmol/L Reserpine, 10 μmol/L Reserpine, 20 μmol/L Reserpine and 50 μmol/L Reserpine. The dual-fluorescence mRFP-GFP-LC3 was used to trace the autophagosomes of RA-VSMC. Real-time fluorescence quantitative PCR (qPCR) and Western blot (WB) were used to detect the expression of LC3 and Beclin1. Results The MTT results showed that compared with the control group, the cell viability in the 5 μmol/L Reserpine group, 10 μmol/L Reserpine group, 20 μmol/L Reserpine group and 50 μmol/L Reserpine group were significantly reduced (P ＜ 0.05 or P ＜ 0.01). The results of the dual-fluorescence mRFP-GFP-LC3 showed that there were no significant autophagosomes in the cells of the control group and the 10 μmol/L Reserpine group, and the 50 μmol/L Reserpine group and EBSS group formed significant autophagosomes. qPCR results showed that compared with the control group, there was no significant difference in LC3 mRNA expression level in the 10 μmol/L Reserpine group and the control group (P ＞ 0.05). LC3 mRNA expression levels in 50 μmol/L Reserpine group and EBSS group increased significantly (P ＜ 0.05 or P ＜ 0.01). Beclin1 mRNA expression levels was significantly increased in 10 μmol/L Reserpine group, 50 μmol/L Reserpine group and EBSS group (P ＜ 0.01). WB results showed that, compared with the control group, the 10 μmol/L Reserpine group, 50 μmol/L Reserpine group and EBSS group LC3-Ⅱ/LC3-Ⅰ ratio increased significantly (P ＜ 0.01), Beclin1 protein expression was significantly increased in 10 μmol/L Reserpine group, 50 μmol/L Reserpine group and EBSS group (P ＜ 0.01). Conclusion Reserpine may regulate the proliferation of vascular smooth muscle cells through autophagy.
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2020, Vol. 17 
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