Ginsenoside Rg1 inhibits apoptosis by inducing autophagy in Raw 264.7 macrophages

LING Lu YANG Ping GAI Sheng-kun LIU Ran CHEN Yuan-li LU Di* SUN Lin*

doi:10.16098/j.issn.0529-1356.2016.05.004

PDF(1060 KB)

Acta Anatomica Sinica ›› 2016, Vol. 47 ›› Issue (5) : 599-606. DOI: 10.16098/j.issn.0529-1356.2016.05.004

Cell and Molecules Biology

Ginsenoside Rg1 inhibits apoptosis by inducing autophagy in Raw 264.7 macrophages

LING Lu¹ YANG Ping² GAI Sheng-kun³LIU Ran¹ CHEN Yuan-li²LU Di ^4* SUN Lin^1*

Author information +

History +

Abstract

Objective To investigate the regulation role and mechanism of ginsenoside Rg1 on autophagy and apoptosis in Raw 264.7 macrophages stimulated with serum free. Methods Raw 264.7 macrophages were cultured and treated differently in vitroand randomly divided into the control group, the serum free group (12, 24, 36, 48 and 60 hours) in which ginsenoside Rg1 (50μmol/L) was added with serum free treatment group (24, 36 and 48 hours), pretreatment with autophagy inhibitor 3-methyladennine (3-MA) (5mmol/L) for 1 hour then added ginsenoside Rg1 (50μmol/L) with serum free treatment group and corresponding control group. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Atg5, Beclin 1, cleaved Caspase-3, Bcl-2 and Bax were detected by Western blotting respectively. The variation of protein expression level of LC3 was measured by double immunofluorescence labeling. The morphology of cell nucleus was measured by Hochest 33342/PI double fluorescent double staining. Results Different time (12, 24, 36, 48 and 60 hours) of the serum free groups induced autophagy. Compared with different time of the serum free groups, the ginsenoside Rg1 (50μmol/L) was added with serum free group up-regulated the protein expression of LC3, Atg 5 and Beclin 1. Compared with the ginsenoside Rg1 (50μmol/L) group, pretreatment with 3-MA (5mmol/L) inhibited the protein expression of Bcl-2 and up-regulated the protein expression of cleaved Caspase-3 and Bax and the quantity of apoptosis in Raw 264.7 macrophages. Conclusion Ginsenoside Rg1 effectively attenuates serum free-induced apoptosis by inducing the autophagy in Raw 264.7 macrophages.

Key words

Ginsenoside Rg1 / Autophagy / Raw 264.7 macrophage / Western blotting / Double immunofluorescent labeling / Hochest 33342/PI fluorescent double staining / Mouse

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

LING Lu YANG Ping GAI Sheng-kun LIU Ran CHEN Yuan-li LU Di SUN Lin. Ginsenoside Rg1 inhibits apoptosis by inducing autophagy in Raw 264.7 macrophages[J]. Acta Anatomica Sinica. 2016, 47(5): 599-606 https://doi.org/10.16098/j.issn.0529-1356.2016.05.004

References

［1］Keebaugh AJ, Sioutas C, Pakbin P, et al. Is atherosclerotic disease associated with organic components of ambient fine particles［J］. Sci Total Environ, 2015, 533(12):69-75.

［2］Seimon T, Tabas I. Mechanisms and consequences of macrophage apoptosis in atherosclerosis［J］. J Lipid Res, 2009, 50(Suppl):S382-387.

［3］Lavandero S, Troncoso R, Rothermel BA, et al. Cardiovascular autophagy: Concepts, controversies and perspectives［J］. Autophagy, 2013, 9(10):1455-1466.

［4］Brichkina A, Bulavin DV. Wiping Out Atherosclerosis with Autophagy［J］. Autophagy, 2012, 8(10):1545-1547.

［5］Quimet M, Franklin V, Mak E, et al. Autophagy regulates cholesterol efflux from macrophage foam cells via lysosomal acid lipase［J］.Cell Metab, 2011, 13(6):655-667.

［6］Huang HY. Pharmacological activity research progress of ginsenoside Rg1［J］. Practical Journal of Traditional Chinese Medicine, 2012, 28(7):608-609. (in Chinese)

黄海英.人参皂苷Rg1药理作用研究进展［J］.实用中医药杂志, 2012, 28(7):608-609.

［7］Endemann DH, Schiffrin EL. Endothelial dysfunction［J］. J Am Soc Nephrol, 2004, 15(8):1983-1992.

［8］Zhu JD, Yu Y, Ge G. Effects of basic fibroblast growth factor on endothelial function and structure of the basilar artery of atherosclerotic rats［J］. Acta Anatomica Sinica, 2014，45(4):469-474. (in Chinese)

朱俊德，余彦，戈果. 碱性成纤维细胞生长因子对动脉粥样硬化大鼠脑基底动脉血管内皮功能与结构的影响［J］.解剖学报，2014，45(4):469-474.

［9］Gautier EL， Huby T, Witzum JL, et al. Macrophage apoptosis exerts divergent effects on atherogenesis as a function of lesion stage［J］. Circulation, 2009, 119(13):1795-1804.

［10］Wang X, Liao D, Lin PH, et al. Highly active antiretroviral therapy drugs inhibit in vitro cholesterol efflux from human macrophage-derived foam cells［J］. Lab Invest, 2009, 89(12):1355-1363.

［11］Yorimitsu T, Klionsky DJ. Autophagy:molecular machinery for self-eating［J］. Cell Death Differ, 2005, 12(Suppl 2):1542-1552.

［12］Kroemer G, Levine B. Autophagic cell death: the story of a misnomer［J］. Nat Revmol Cell Bio, 2008, 9(12):1004-1010.

［13］Liao X, Sluimer J C, Wang Y, et al. Macrophage autophagy plays a protective role in advanced atherosclerosis［J］. Cell Metab, 2012, 15(4):545-553.

［14］Li G, Scull C, Ozcan L, et al. NADPH oxidase links endoplasmic reticulum stress, oxidative stress, and PKR activation to induce apoptosis［J］. J Cell Biol, 2010, 191(6):1113-1125.