Protective Effects of Dohongsamul-tang on 2-deoxy-D-glucose Induced Autophagic Cell Death in C6 Glial Cells

도홍사물탕(挑紅四物湯)이 C6 신경교세포의 2-DG에 의한 오토파지성 세포사멸에 미치는 영향

  • Shin, Hak-Soo (Department of Third Medicine, Professional Graduate School of Oriental Medicine, WonkWang University) ;
  • Lee, Seung-Geun (Department of Internal Medicine, School of Oriental Medicine, WonkWang University) ;
  • Moon, Byung-Soon (Department of Third Medicine, Professional Graduate School of Oriental Medicine, WonkWang University)
  • 신학수 (원광대학교 한의학전문대학원 제3의학과) ;
  • 이성근 (원광대학교 한의과대학 내과학교실) ;
  • 문병순 (원광대학교 한의학전문대학원 제3의학과)
  • Published : 2009.06.25

Abstract

The water extract of Dohongsamul-tang(DHSMT) has been traditionally used in treatment of ischemic heart and brain diseases in Oriental Medicine. However, little is known about the mechanism by which DHSMT protects C6 glial cells from glucose deprevation induced damages. Therefore, this study was designed to evaluate the protective effects of DHSMT on 2-deoxy-D-glucose induced autophagy of C6 glial cells. Autophagic phenotype is evaluated by fluorescence microscopy and flow cytometry with specific biological staining dyes, including monodansylcadaverine and acridine orange, as well as Western blot analysis with microtubule-associated protein 1 light chain 3(LC3) and Beclin-1. Treatment with 2-deoxy-D-glucose significantly resulted in a decrease of the viability of C6 glial cells and increase of the extracellular LDH release in a dose and time-dependent manner. However, pretreatment with DHSMT protected C6 glial cells from glucose deprivation with 2-deoxy-D-glucose. The author also observed the fact that autophagy phenotype occurred by 2-deoxy-D-glucose in C6 glial cells. Pretreatment with 3-MA, a pharmacological inhibitior of autophagy, abolished the formation of acidic vesicle organelle in C6 glial cells treated with 2-deoxy-D-glucose. However, pretreatment with DHSMT inhibited the formation of autophagic phenotypes, including formation of acidic vesicle organelle, and increase of the expression of LC-3 II Beclin-1 proteins in C6 glial cells treated with 2-deoxy-D-glucose. Taken together, these data suggest that DHSMT is able to protect C6 glial cells from glucose deprivation with marked inhibition of autophagy formation.

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