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Neuroprotective Effect of Rice with Phellinus linteus Mycelium in HT22 Cells

상황버섯균사체 쌀의 HT22 신경세포 보호 효과

  • Kim, Ji Hyun (Department of Food & Nutrition, Sunchon National University) ;
  • Chun, Soon Sil (Department of Food & Nutrition, Sunchon National University)
  • 김지현 (순천대학교 식품영양학과) ;
  • 전순실 (순천대학교 식품영양학과)
  • Received : 2017.02.27
  • Accepted : 2017.06.13
  • Published : 2017.07.31

Abstract

In this study, the protective effect of rice with Phellinus linteus mycelium (PLMR) against hydrogen peroxide-induced oxidative stress was assessed in a mouse hippocampal neuronal HT22 cell line through (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) salt (MTS) assay and western blot. MTS assay using HT22 cells showed that PLMR extract did not affect viability at a concentration range from 1 mg/mL to 5 mg/mL. However, at concentrations over 10 mg/mL, PLMR extract resulted in increased cell death. Cell viability of HT22 was significantly reduced by $H_2O_2$ treatment, and reduction of cell viability was efficiently restored by treatment with PLMR extract in a dose-dependent manner from 0.1 to 1 mg/mL. Cells treated with $H_2O_2$ showed increased expression of Bax, a pro-apoptotic protein, which was down-regulated by treatment with PLMR extract. On the other hand, cells treated with $H_2O_2$ resulted in reduced expression of Bcl-2, an anti-apoptotic protein, which was restored by treatment with PLMR extract. In addition, treatment with PLMR extract reduced expression of cleaved caspase 3 and PARP, which were up-regulated by $H_2O_2$ treatment. The results may suggest that treatment with PLMR extract would suppress $H_2O_2$-induced apoptosis of HT22 cells.

상황버섯균사체 쌀의 뇌세포 보호 효과를 분석하여 상황버섯균사체 쌀에 대한 기초자료를 제공하고, 이를 이용하여 뇌 질환의 예방과 관련된 기능성 식품으로의 개발가능성을 찾고자 하였다. 상황버섯균사체 쌀 추출물을 처리하여 마우스 해마세포주 HT22의 세포보호 효과를 알아보기 위해 MTS assay를 이용한 세포생존율과 western blot을 이용한 세포사멸 단백질의 발현을 관찰하였다. 상황버섯균사체 쌀 추출물의 단일 독성을 측정한 결과 상황버섯균사체 쌀 추출물 처리군은 102.68% 이상의 생존율을 보여 신경세포의 사멸에 영향을 주지 않았다. 또한, 상황버섯균사체 쌀 추출물과 $H_2O_2$를 함께 처리하였을 때 $H_2O_2$로 자극된 세포는 63.80%의 세포생존율을 나타내었고, 상황버섯균사체 쌀 추출물과 $H_2O_2$가 함께 처리된 세포는 76.85%, 92.46%, 95.57%로 농도 의존적으로 세포생존율이 증가하였다. HT22 cell에 $H_2O_2$와 상황버섯균사체 쌀 추출물을 처리하여 apoptosis 유도 단백질을 측정한 결과, pro-apoptotic protein인 Bax는 발현이 억제되었고, anti-apoptotic protein인 Bcl-2는 발현이 증가함을 확인하였다. 또한, caspase-3와 PARP의 발현을 억제하여 상황버섯균사체 쌀 추출물이 $H_2O_2$로 유도되는 apoptosis를 억제하는 것을 확인할 수 있었다.

Keywords

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