CAJ | 학술논문

目的观察模拟缺血再灌注(IR)培养乳兔窦房结细胞(SANC)损伤及左旋精氨酸(L-Arg)的保护作用.方法对SANC模拟低糖缺氧培养,添加L-Arg和L-Arg 加 L-NAME(L-Arg抑制剂)与培养细胞共同孵育,吖啶橙(AO)标记细胞凋亡,检测细胞内过氧化物歧化酶(SOD)活性、丙二醛(MDA)、总一氧化氮合酶(NOS)、氧化亚氮(NO)水平.细胞分为正常对照组、I120R120 组、I120R120加L-Arg组、I120R120加L-Arg加L-NAME组4组.结果凋亡细胞体积明显缩小,核呈黄绿色,断裂为多个碎块状,由膜包裹着凸起于细胞表面呈黄绿色凋亡小体.与I120R120组凋亡率[(5.21±1.59)%]比较,I120R120加L-Arg组凋亡率[(8.70±3.10)%]显著降低(P<0.01);I120R120加L-Arg组细胞SOD活性[(46 820±14 450) U/g]较I120R120组 [(25 030±8 440) U/g]显著上升,而I120R120加L-Arg组MDA[(5.55±3.71)mmol/g]、NO[(3.65±1.02) U/g]和NOS[(3.73±0.24) μmol/L]较I120R120组MDA[(8.42±4.21)mmol/g]、NO[ (4.62±1.20) U/g]和NOS[(4.96±0.52) μmol/L]显著下降(Pa<0.05);而I120R120加L-Arg加L-NAME组各指标较I120R120组无显著性差异(Pa>0.05).结论补充NO合成底物L-Arg能清除自由基,增加SOD活性,增强细胞抗氧化损伤能力,可能是阻断氧自由基所介导的细胞凋亡机制之一.
목적 관찰모의결혈재관주(IR)배양유토두방결세포(SANC)손상급좌선정안산(L-Arg)적보호작용.방법 대SANC모의저당결양배양,첨가L-Arg화L-Arg 가 L-NAME(L-Arg억제제)여배양세포공동부육,아정등(AO)표기세포조망,검측세포내과양화물기화매(SOD)활성、병이철(MDA)、총일양화담합매(NOS)、양화아담(NO)수평.세포분위정상대조조、I120R120 조、I120R120가L-Arg조、I120R120가L-Arg가L-NAME조4조.결과 조망세포체적명현축소,핵정황록색,단렬위다개쇄괴상,유막포과착철기우세포표면정황록색조망소체.여I120R120조조망솔[(5.21±1.59)%]비교,I120R120가L-Arg조조망솔[(8.70±3.10)%]현저강저(P<0.01);I120R120가L-Arg조세포SOD활성[(46 820±14 450) U/g]교I120R120조 [(25 030±8 440) U/g]현저상승,이I120R120가L-Arg조MDA[(5.55±3.71)mmol/g]、NO[(3.65±1.02) U/g]화NOS[(3.73±0.24) μmol/L]교I120R120조MDA[(8.42±4.21)mmol/g]、NO[ (4.62±1.20) U/g]화NOS[(4.96±0.52) μmol/L]현저하강(Pa<0.05);이I120R120가L-Arg가L-NAME조각지표교I120R120조무현저성차이(Pa>0.05).결론 보충NO합성저물L-Arg능청제자유기,증가SOD활성,증강세포항양화손상능력,가능시조단양자유기소개도적세포조망궤제지일.
Objective To observe the protective effect of L-Arginine (L-Arg) on simulated ischemia reperfusion (IR) injury of cultured sinoatrial node cell (SANC) in neonatal rabbits.Methods The apoptotic cells were stained with acridine orange(AO) fluorescence and the contents of superoxide(SOD),malondialdehyde(MDA),nitric oxide synthase(NOS),and nitric oxide(NO) of the cultured cells in the different groups were examined.SANC were cultured in hypoglycemic and hypoxic medium to simulate IR injury.Then,L-Arg and L-Arg plus L-nitro-Arginine methyl ester(L-NAME,L-Arg inhibitor) were added in it.SANC were set into 4 groups: control group,I120R120 group,I120R120 plus L-Arg group and I120R120 plus L-Arg plus L-NAME group.Results AO-fluorescence staining showed that the apoptotic cell was dwindled and the nucleus was fractured into many small shivers which were enwrapped and heaved on cell′s surface to become yellow green apoptotic body.The rate of apoptosis was significantly decreased in I120R120 plus L-Arg group [(5.21±1.59)%]compared with I120R120 group [(8.70±3.10)%](P<0.01);The activity of SOD was increased significantly in I120R120 plus L-Arg group[(46 820±14 450)U/g]compared with I120R120group [(25 030±8 440)U/g]while the MDA[(5.55±3.71)mmol/g],NO[(3.65±1.02)μmol/L]and NOS [(3.73±0.24)U/g]were significantly decreased in I120R120 plus L-Arg group compared with I120R120group[(8.42±4.21)mmol/g,(4.62±1.20)μmol/L,(4.96±0.52)U/g](Pa<0.05).However,there were no significant differences in them between I120R120 plus L-Arg plus L-NAME group and I120R120 group (Pa>0.05).Conclusions The supplement of appropriate amount of L-Arg can clean out and decrease the excrescent oxygen free radicals,improve SOD activity and increase the oxidation resistance of cells,which may be one of the mechanisms interdicting the apoptotic process induced by oxygen free radicals.