CAJ | 학술논문

目的比较观察甲基胍、1-甲脲乙醇酸酐对人肾小管上皮细胞的毒性作用.方法 HK-2细胞作为研究对象,分为三组培养:正常对照(A)组,甲基胍(B)组、1-甲脲乙醇酸酐(C)组,用MTT比色试验法检测细胞增殖,NAG酶释放检测对细胞的毒性,Hoechst染色及Annexin V-FITC/PI流式细胞术检测凋亡.结果 HK-2细胞加入甲基胍、1-甲脲乙醇酸酐后,较正常对照组吸光度值显著下降(0.188±0.011,0.176±0.010 vs 0.545±0.021,F＝1557.74,P＜0.01),NAG酶浓度显著上升(20.488±0.473,22.225±0.565 vs 5.125±0.198,F＝3848.22,P＜0.01).Hoechst33258染色显示凋亡细胞呈现核固缩,染色加深,并出现凋亡小体.流式细胞术显示B组、C组HK-2细胞凋亡率显著高于A组(18.23±1.1581,20.22±1.1433 vs 2.473±0.321,F＝526.06,P＜0.01).C组吸光度值显著低于B组(0.176±0.010 vs 0.188±0.011,t＝2.26,P＜0.05),NAG酶浓度显著高于B组(22.225±0.565 vs 20.488±0.473,t＝-6.67,P＜0.01),凋亡率显著高于B组(20.22±1.1433 vs 18.23±1.1581,t＝-2.762,P＜0.05).结论 1-甲脲乙醇酸酐对肾小管上皮细胞的毒性作用强于甲基胍.
목적 비교관찰갑기고、1-갑뇨을순산항대인신소관상피세포적독성작용.방법 HK-2세포작위연구대상,분위삼조배양:정상대조(A)조,갑기고(B)조、1-갑뇨을순산항(C)조,용MTT비색시험법검측세포증식,NAG매석방검측대세포적독성,Hoechst염색급Annexin V-FITC/PI류식세포술검측조망.결과 HK-2세포가입갑기고、1-갑뇨을순산항후,교정상대조조흡광도치현저하강(0.188±0.011,0.176±0.010 vs 0.545±0.021,F＝1557.74,P＜0.01),NAG매농도현저상승(20.488±0.473,22.225±0.565 vs 5.125±0.198,F＝3848.22,P＜0.01).Hoechst33258염색현시조망세포정현핵고축,염색가심,병출현조망소체.류식세포술현시B조、C조HK-2세포조망솔현저고우A조(18.23±1.1581,20.22±1.1433 vs 2.473±0.321,F＝526.06,P＜0.01).C조흡광도치현저저우B조(0.176±0.010 vs 0.188±0.011,t＝2.26,P＜0.05),NAG매농도현저고우B조(22.225±0.565 vs 20.488±0.473,t＝-6.67,P＜0.01),조망솔현저고우B조(20.22±1.1433 vs 18.23±1.1581,t＝-2.762,P＜0.05).결론 1-갑뇨을순산항대신소관상피세포적독성작용강우갑기고.
Objective To investigate the effect of methylguanidine and 1-methylhydantoin on cells cytotoxicity, apoptosis in human renal tubular epithelial cell line (HK-2). Methods Human PTEC cell line HK-2 was used in this study. HK-2 was cultured and divided into 3 groups: Norma1 control group (A), methylguanidine group(B) and 1-methylhydantoin group (C). The cell inhibitory rate of HK-2 was detected by MTT method. The cytotoxicity of methylguanidine to HK-2 was determined by NAG release test. Cell apoptosis was evaluated by using Hoechst stain and FACS with Annexin-V/PI. Results The OD value and NAG concentration of creatinine, methylguanidine and 1-methylhydantoin group were compared with normal control group. OD value decreased and NAG concentration significantly increased(0.188±0.011, 0.176±0.010 vs 0.545±0.021, F＝1557.74, P＜0.01; 20.488±0.473, 22.225±0.565 vs 5.125±0.198, F＝3848.22, P＜0.01). By Hoechst stain, pycnosis and apoptotic body could be found when HK-2 was cultivated in methylguanidine 1-methylhydantoin group. In methylguanidine, 1-methylhydantoin group apoptotic HK-2 apparently increased, compared with that in control group (18.23±1.1581, 20.22±1.1433 vs 2.473±0.321, F＝526.06, P＜0.01). Compared with group B, the OD value in group C decreased significantly (0.176±0.010 vs 0.188±0.011,t＝2.26, P＜0.05), NAG concentration increased significantly (22.225±0.565 vs 20.488±0.473,t＝-6.67, P＜0.01), and apoptotic rate in-creased significantly (20.22±1.1433 vs 18.23±1.1581,t＝-2.762, P＜0.05). Conclusions 1-methylhydantoin has more powerful cytotoxic effect to renal tubular epithelial cells than that of Methylguanidine.