CAJ | 학술논문

目的探讨醋酸铅对人肾小管上皮细胞(HK-2)凋亡的作用及对超微结构的影响。方法经5、10、20 μmol/L醋酸铅染毒HK-2细胞24h后，Hochest33342染色法观察细胞形态学变化，透射电子显微镜下观察细胞超微结构的变化，分光光度计检测细胞培养上清中乳酸脱氧酶(LDH)活力、丙二醛(MDA)含量，DNA Ladder实验检测醋酸铅对HK-2基因组的影响，应用流式细胞仪检测细胞凋亡率。结果 Hochest33342-PI染色显示，染铅组细胞呈凋亡形态学改变。透射电子显微镜下观察发现，染铅组细胞胞质内出现空泡，核同缩，核膜结构模糊，并出现凋亡小体。染铅组细胞培养上清中LDH活力及MDA含量较对照组明显升高，差异有统计学意义(P＜0.01)。HK-2基因结果显示，染铅组出现较为明显的DNA损伤现象。5、10、20 μmol/L染铅组细胞凋亡率分别为14.16％±2.94％、19.45％±2.73％、25.01％±3.97％，均明显高于对照组(5.81％±2.18％)，差异均有统计学意义(P＜0.05)。结论醋酸铅可促进HK-2细胞凋亡，进而影响肾脏正常功能。
목적 탐토작산연대인신소관상피세포(HK-2)조망적작용급대초미결구적영향。방법 경5、10、20 μmol/L작산연염독HK-2세포24h후，Hochest33342염색법관찰세포형태학변화，투사전자현미경하관찰세포초미결구적변화，분광광도계검측세포배양상청중유산탈양매(LDH)활력、병이철(MDA)함량，DNA Ladder실험검측작산연대HK-2기인조적영향，응용류식세포의검측세포조망솔。결과 Hochest33342-PI염색현시，염연조세포정조망형태학개변。투사전자현미경하관찰발현，염연조세포포질내출현공포，핵동축，핵막결구모호，병출현조망소체。염연조세포배양상청중LDH활력급MDA함량교대조조명현승고，차이유통계학의의(P＜0.01)。HK-2기인결과현시，염연조출현교위명현적DNA손상현상。5、10、20 μmol/L염연조세포조망솔분별위14.16％±2.94％、19.45％±2.73％、25.01％±3.97％，균명현고우대조조(5.81％±2.18％)，차이균유통계학의의(P＜0.05)。결론 작산연가촉진HK-2세포조망，진이영향신장정상공능。
Objective To explore the toxic effects of lead acetate on the apoptosis and ultrastructure of human renal tubular epithelial cells (HK-2). Methods After HK-2 cells were exposed to 5, 10 and 20 μ mol/L lead acetate for 24 h, the morphological changes of HK-2 cells were observed by Hochest 33342-PI staining, and the uitrastructure changes of HK-2 cells were examined under a electron microscope, LDH activity and MDA content in supernatant of HK-2 cellular culture were detected by spectrophotometer, DNA damage of HK-2 was determined by DNA ladder and the apoptotic rates of HK-2 cells were measured by flow cytometry. Results The morphological changes of apoptotic HK-2 cells in exposure group were observed by Hochest 33342-PI staining. The cytoplasm vacuoles, karyopycnosis, nuclear membrane vague and apoptotic bodies in HK-2 cells of exposure group were found under electron microscopy. LDH activity and MDA contents in exposure group increased significantly, as compared to control group (P＜0.01). The results of DNA Ladder showed that DNA damage of HK-2 cells in exposure group appeared. The apoptotic rates of HK-2 cells exposed to 5, 10, 20 μmol/L lead acetate were 14.16％ ± 2.94％, 19.45％ ± 2.73％, 25.01％ ± 3.97％, respectively, which were significantly higher than that (5.8 1 ％±2.18％) in control group (P＜0.05). Conclusion Lead acetate could remarkably induce the apoptosis of HK-2 cells and affect the kidney.