ラット頬粘膜および舌のミトコンドリア分離法と細胞障害物質のミトコンドリア呼吸調節に及ぼす影響

概要

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Aim of the study is to purify the intact mitochondria, having high respiratory activity, from buccal mucosa and tongue of rats. Mitochondria were prepared under two conditions; that is, homogenizing duration and alkaline proteinase concentration. The optimal conditions for purification were evaluated by the ADP/O ratio, respiratory control ratio (RCR), and succinate dehydrogenase (SDH) activity. Both ADP/O ratio and RCR values showed maximum in mitochondrial preparations of both tissues, when they were prepared with following conditions; homogenizing duration of 10s in alkaline proteinase of 5mg/g wet wt. The quality of purified mitochondria was evaluated by a specific activity of SDH. Results of SDH measurements showed that the highest activity was present in the mitochondrial fraction prepared by the same condition. Furthermore, it was also examined the possibility to use ADP/O ratio and RCR as a parameter for cell damage. The effects of E.coli lipopolysaccharide (EcLPS) and leukotoxin (Lx) on the present preparations were compared with those of heart and gingival mitochondria. Both EcLPS and Lx reduced ADP/O ratio and RCR in all mitochondria in a dose-dependent manner. These results indicate that an optimal method for mitochondria, having sufficient respiratory activity, was determined to monitor the cell damage in the tissues.
福岡歯科大学学会の論文
2002-12-20