Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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The Role of Helicobacter pylori's Fur Protein in the Oxidative Stress Induced by Photodynamic Therapy

Photodynamic Therapy에 의한 산화적 스트레스 조건에서 Helicobacter pylori의 Fur 단백질의 역할

  • Received : 2011.04.20
  • Accepted : 2011.06.15
  • Published : 2011.06.30
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Abstract

The role of the ferric uptake regulator (Fur) of Helicobacter pylori in the oxidative stress was investigated in this study. A fur knockout mutant of H. pylori was constructed by replacing the fur gene with an aphA (kanamycin resistant marker) gene. Photodynamic therapy using methylene blue (MB) and 660 nm light was chosen to induce oxidative stress. The bactericidal effect of photodynamic therapy (PDT) was compared between wild type H. pylori and fur knockout mutant H. pylori. The degree of oxidative damage of DNA was confirmed using alkaline gel electrophoresis and an assay of 8-hydroxy-2-deoxyguanosine (8-OHdG). In control groups, the number of viable cells was maintained constantly during experiment. After PDT, the mutant H. pylori showed 10,000 times decreased viable cell number compared with wild type H. pylori. Depending on the exposure time of 660 nm light, the 3-fold increase in the concentration of 8-OHdG was observed in mutant H. pylori. The results of this study showed that H. pylori's Fur protein may play a role in oxidative stress induced by PDT.

Helicobacter pylori균의 ferric uptake regulator (Fur) 단백질의 산화적 손상에 대한 역할을 연구하였다. H. pylori균의 fur 유전자를 제거한 돌연변이체를 만들고 wild type H. pylori균과 돌연변이체 균의 산화적 스트레스에 대한 반응을 비교하였다. 산화적 스트레스는 methylene blue와 660 nm 파장의 빛을 이용하는 광역학적 치료방법으로 유도하였다. 산화적 스트레스를 가한 실험조건에서 wt H. pylori와 돌연변이체의 생존력, DNA 손상의 정도를 비교 검토하였다. 그 결과 fur 유전자가 제거된 돌연변이체의 생균수가 wt에 비해 10,000배 가량 감소한 것을 알 수 있었으며 DNA의 산화적 손상의 marker인 8-hydroxy-2-deoxyguanosine (8-OHdG)의 양도 fur 유전자 제거된 돌연변이에서 wild type에 비해 3배 정도 더 생성됨을 확인하였다. 따라서 본 실험결과 H. pylori균의 fur 유전자가 PDT법으로 유도한 산화적 스트레스에 방어 기작을 하는 것으로 사료된다.

Keywords

References

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