補充葉酸對大白鼠初代肝細胞經氧化劑處理後粒線體質量與膜電位的影響

Effect of Folate Supplementation on Mitochondrial Mass and Membrane Potential of Rat Primary Hepatocytes Treated with Oxidants

盧信得(Hsin-Te Lu) ； 李宗貴(Chong-Kuei Lii) ； 鄭心嫻(Hsing-Hsien Cheng) ； 許瑞芬(Rwei-Fen S. Huang)

《中華民國營養學會雜誌》 29卷1期 (2004/03) Pp. 38-47

https://doi.org/10.6691/JCNS.200403_29(1).0005

摘要

本研究探討葉酸補充對氧化壓力所誘發之肝細胞粒線體傷害的影響。先以兩階段膠原蛋白酶灌流法分離大白鼠初代肝細胞，培養於不同葉酸含量的培養液中，再以氧化劑同半胱胺酸、銅離子與menadione處理，觀察補充葉酸對大白鼠初代肝細胞經氧化劑處理後粒線體質量與膜電位的影響。研究結果顯示，同半胱胺酸處理初代肝細胞24小時後，粒線體質量與粒線體膜電位顯著下降。補充葉酸顯著改善初代肝細胞因同半胱胺酸所誘發的粒線體膜電位下降情況，但無法改善肝細胞粒線體質量下降現象。以銅離子處理初代肝細胞24小時後，大白鼠初代肝細胞粒線體膜電位顯著降低，補充葉酸能顯著減緩粒線體傷害性。以氧化劑menadione處理初代肝細胞1小時後，會急速引起細胞變形與死亡，並誘發粒線體質量與粒線體膜電位顯著降低。葉酸的補充並不能改善menadione所誘發的粒線體質量與粒線體膜電位降低。由此可知，在menadione急性氧化壓力誘發下，補充葉酸對肝細胞粒線體無保護性。葉酸補充對同半胱胺酸與銅離子氧化劑所誘發的粒線體膜電位去極化有顯著改善性。

關鍵字

葉酸補充 ； 大白鼠初代肝細胞 ； 粒線體質量 ； 粒線體膜電位 ； 氧化劑

並列摘要

The aim of this study was to investigate the effects of folate supplementation on mitochondrial mass and membrane potential of rat primary hepatocytes treated with oxidants. Rat primary hepatocytes were obtained by a two-step collagenase perfusion and incubation in medium containing various concentrations of folate (0-1000 μM). Cells were treated with homocysteine(Hcy), Cu++, or menadione, and changes of mitochondrial mass as well as membrane potential were assessed by flow cytometry. After Hcy treatment for 24 h, mitochondrial mass and membrane potential had significantly decreased in rat primary hepatocytes. Folate supplementation could partially normalize the mitochondrial membrane potential of Hcy-treated cells, but did not affect changes in mitochondrial mass. Upon Cu++ treatment, the mitochondrial membrane potential of rat primary hepatocytes was significantly decreased. The decrease in the mitochondrial membrane potential induced by copper ion, was ameliorated by folate supplementation. After menadione treatment, cells became morphologically deformed and began dying. Folate supplementation affect neither the mitochondrial mass nor membrane potential changes in menadione-treated cells. Taken together, folate supplementation has some protective effects on Cu++, and Hcy-induced changes in the mitochondrial membrane potential, but failed to affect mitochondrial mass.

並列關鍵字

folate supplementation ； rat primary hepatocytes ； mitochondrial mass ； mitochondrial mem-brane potential ； oxidants