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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 |
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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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[1] Feil S,Fehrenbacher B,Lukowski R,et al. Transdifferentiation of vascular smooth muscle cells to macrophage-like cells during atherogenesis [J]. Circ Res,2014,115(7):662-667.
[2] Yang J,Li X,Al-Lamki RS,et al. Sildenafil potentiates bone morphogenetic protein signaling in pulmonary arterial smooth muscle cells and in experimental pulmonary hypertension [J]. Arterioscler Thromb Vasc Biol,2013,33(1):34-42.
[3] Jaminon A,Reesink K,Kroon A,et al. The Role of Vascular Smooth Muscle Cells in Arterial Remodeling: Focus on Calcification-Related Processes [J]. Int J Mol Sci,2019,20(22).pii:E5694.
[4] Saha S,Panigrahi DP,Patil S,et al. Autophagy in health and disease: A comprehensive review [J]. Biomed Pharmacother,2018,104:485-495.
[5] Leao AH,Sarmento-Silva AJ,Santos JR,et al. Molecular,Neurochemical,and Behavioral Hallmarks of Reserpine as a Model for Parkinson′s Disease:New Perspectives to a Long-Standing Model [J]. Brain Pathol,2015,25(4):377-390.
[6] 王奥昉.缬沙坦与利血平治疗高血压的疗效研究[J].中国地方病防治杂志,2017,32(4):440.
[7] 王夏青.5-羟色胺2C受体激动剂抗精神分裂症作用的潜在机制[D].重庆:西南大学,2018.
[8] 黄桢钧,刘彬,刘本荣,等.双荧光mRFP-eGFP-LC3体系在细胞自噬中的作用[J].贵阳医学院学报,2015,40(10):1029-1032.
[9] Shi W,Zhou Y,Wang H,et al. Synergistic interaction of hypertension and hyperhomocysteinemia on chronic kidney disease:Findings from the National Health and Nutrition Examination Survey 1999-2006 [J]. J Clin Hypertens (Greenwich),2019,21(10):1567-1577.
[10] Lee KI,Kim MJ,Koh H,et al. The anti-hypertensive drug reserpine induces neuronal cell death through inhibition of autophagic flux [J]. Biochem Biophys Res Commun,2015,462(4):402-408.
[11] 蔡美丽,徐健飞.高血压与心血管重构相关关系的研究进展[J].现代医学与健康研究电子杂志,2018,2(8):131,137.
[12] Smith M,Wilkinson S. ER homeostasis and autophagy [J]. Essays Biochem,2017,61(6):625-635.
[13] Cadwell K. Crosstalk between autophagy and inflammatory signalling pathways:balancing defence and homeostasis [J]. Nat Rev Immunol,2016,16(11):661-675.
[14] Um JH,Yun J. Emerging role of mitophagy in human diseases and physiology [J]. BMB Rep,2017,50(6):299-307.
[15] Rybstein MD,Bravo-San pedro JM,Kroemer G,et al. The autophagic network and cancer [J]. Nat Cell Biol,2018,20(3):243-251.
[16] Ma Y,Galluzzi L,Zitvogel L,et al. Autophagy and cellular immune responses [J]. Immunity,2013,39(2):211-27.
[17] Liu C,Xu P,Chen D,et al. Roles of autophagy-related genes Beclin-1 and LC3 in the development and progression of prostate cancer and benign prostatic hyperplasia [J]. Biomed Rep,2013,1(6):855-860.
[18] Xu F,Fang Y,Yan L,et al. Nuclear localization of Beclin 1 promotes radiation-induced DNA damage repair independent of autophagy [J]. Sci Rep,2017,7:45385.
[19] Wang RC,Wei Y,An Z,et al. Akt-mediated regulation of autophagy and tumorigenesis through Beclin 1 phosphorylation [J]. Science,2012,338(6109):956-959.
[20] Tai S,Hu XQ,Peng DQ,et al. The roles of autophagy in vascular smooth muscle cells [J]. Int J Cardiol,2016, 211:1-6.
[21] Ibrahim YF,Shults NV,Rybka V,et al. Docetaxel Reverses Pulmonary Vascular Remodeling by Decreasing Autophagy and Resolves Right Ventricular Fibrosis [J]. J Pharmacol Exp Ther,2017,363(1):20-34. |
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