Carnosine 與anserine 對於亞鐵離子、銅離子及過氧化氫誘導淋巴球細胞DNA氧化損傷之影響   全文下載

Inhibitory Effect of Carnosine and Anserine on DNA Oxidative Damage Induced by Fe2+, Cu2+ and H2O2 in Lymphocytes

本研究以彗星試驗法探討carnosine及anserine對不同誘發劑造成人體淋巴球細胞DNA氧化傷害之影響。結果顯示carnosine與anserine在100μM對人體淋巴球細胞有輕微的基因毒性及細胞急性，但細胞與carnosine與anserine混和反應30分鐘後，控制組（0μM）細胞平均tail DNA為4.3%，carnosine與anserine在濃度5-100μM的範圍內tail DNA分別為4.7-7.3%及6.1-10.0%且細胞存活率皆在85%以上。在加入誘發淋巴球細胞DNA氧化傷害之試劑-Fe2+及Cu2+的試驗中，結果顯示Fe2+及Cu2+對淋巴球細胞有明顯的細胞毒性及DNA氧化傷害，且隨著劑量的增加對淋巴球細胞DNA損傷之情形愈嚴重。Fe2 +濃度為750μM時tail DNA高達36.7%，明顯的與濃度50-500μM時的tail DNA（15.3-22.9%）有顯著性的差異（P < 0.05）。在Cu2+的試驗中，結果顯示濃度為25μM時tail DNA已達38.1%。在carnosine與anserine對不同誘發劑，Fe2+、Cu2+及H2O2，對淋巴球細胞DNA氧化傷害的試驗中發現，carnosine或anserine先與Fe2+或Cu2+反應30分鐘後再與淋巴球細胞反應，結果顯示有抑制細胞DNA氧化傷害的效果，在100μM的濃度下約有50%的抑制率。然而carnosine或anserine與H2O2同時與淋巴球細胞反應其抑制效果分別為46.4%及49.3%。Carnosine及anserine抑制Fe2+及Cu2+所誘導的淋巴球DNA氧化傷害較H2O2所誘導的明顯，其可能的原因是carnosine或anserine來不及清除H2O2，而H2O2已進入細胞造成DNA的氧化傷害。

The effects of carnosine and anserine on oxidative DNA damage in human lymphocytes induced by Fe2+, Cu2+ or H2O2 were investigated using single cell gel electrophoresis (comet assay). Carnosine and anserine induced a slight DNA damage in human lymphocytes after incubation with cells for 30 min. Carnosine and anserine caused a tail DNA % from 4.7% to 7.3% and 6.1% to 10.0% respectively, at a concentration of 5-100 mM. The cell viability of carnosine and anserine to human lymphocytes was more than 85%. Fe2+ and Cu2+ caused a marked cytotoxicity and DNA damage in human lymphocytes with a concentration dependent manner. At a concentration of 100 μM, carnosine and anserine possessed 60-70% inhibitory effect on DNA damage in human lymphocytes when they reacted with Fe2+ or Cu2+ for 30 min before incubated with human lymphocytes. Fe2+, Cu2+, and H2O2-induced DNA damage in human lymphocytes were inhibited by carnosine and anserine in a concentration dependent manner (5-100 μM). However, the inhibitory effect of carnosine and anserine on oxidative DNA damage in human lymphocyte was 46.4% and 49.3%, respectively, when reacted simultaneously with H2O2. Carnosine and anserine possessed more marked inhibitory effect on oxidative DNA damage in human lymphocyte induced by Fe2+ and Cu2+ than that induced by H2O2. This result might be due to carnosine and anserine did not possess significantly scavenging effect on H2O2, thus, H2O2 entered the cell and caused DNA damage. 本研究以彗星試驗法探討carnosine及anserine對不同誘發劑造成人體淋巴球細胞DNA氧化傷害之影響。結果顯示carnosine與anserine在100μM對人體淋巴球細胞有輕微的基因毒性及細胞急性，但細胞與carnosine與anserine混和反應30分鐘後，控制組（0μM）細胞平均tail DNA為4.3%，carnosine與anserine在濃度5-100μM的範圍內tail DNA分別為4.7-7.3%及6.1-10.0%且細胞存活率皆在85%以上。在加入誘發淋巴球細胞DNA氧化傷害之試劑-Fe2+及Cu2+的試驗中，結果顯示Fe2+及Cu2+對淋巴球細胞有明顯的細胞毒性及DNA氧化傷害，且隨著劑量的增加對淋巴球細胞DNA損傷之情形愈嚴重。Fe2 +濃度為750μM時tail DNA高達36.7%，明顯的與濃度50-500μM時的tail DNA（15.3-22.9%）有顯著性的差異（P < 0.05）。在Cu2+的試驗中，結果顯示濃度為25μM時tail DNA已達38.1%。在carnosine與anserine對不同誘發劑，Fe2+、Cu2+及H2O2，對淋巴球細胞DNA氧化傷害的試驗中發現，carnosine或anserine先與Fe2+或Cu2+反應30分鐘後再與淋巴球細胞反應，結果顯示有抑制細胞DNA氧化傷害的效果，在100μM的濃度下約有50%的抑制率。然而carnosine或anserine與H2O2同時與淋巴球細胞反應其抑制效果分別為46.4%及49.3%。Carnosine及anserine抑制Fe2+及Cu2+所誘導的淋巴球DNA氧化傷害較H2O2所誘導的明顯，其可能的原因是carnosine或anserine來不及清除H2O2，而H2O2已進入細胞造成DNA的氧化傷害。