CAJ | 학술논문

目的探讨钙激活钾通道(KCa)在石英致大鼠肺泡巨噬细胞损伤中的作用.方法大鼠肺泡巨噬细胞(NR8383)为受试细胞;以100 μg/ml晶体和无定形石英分别染尘,为晶体石英组和无定形石英组;不施加任何处理,正常培养的细胞为阴性对照组.进行体外实验.首先比较2种石英各自的作用,然后在100 μg/ml晶体石英的基础上加入不同剂量3种类型的KCa特异性阻断剂(BK阻断剂Paxilline、IK阻断剂Tram-34、SK阻断剂Apamin),观察通道阻断剂对石英致细胞损伤的影响.测定细胞存活率、乳酸脱氢酶(LDH)活力、白介素-1β(IL-1β)以及肿瘤坏死因子(TNF-α)释放量.结果与阴性对照组比较,无定形石英组和晶体石英组细胞存活率降低,LDH活力升高,IL-1β和TNF-α释放量增多;与无定型石英组比较,晶体石英组细胞存活率明显降低,LDH活力、IL-1β、TNF-α释放量明显升高,差异均有统计学意义(P＜0.01).与晶体石英组比较,IK阻断剂组细胞存活率增高,差异有统计学意义(P＜0.01);与晶体石英组比较,BK、IK、SK阻断剂组细胞LDH活力降低;BK、IK、SK阻断剂组IL-1β释放量减少,BK、IK阻断剂组TNF-α分泌量减少,差异均有统计学意义(P＜0.05).Tram-34降低IL-1β和TNF-α释放量呈剂量-效应关系.结论阻断大鼠肺泡巨噬细胞KCa可降低石英所致细胞膜破坏,减少炎性因子IL-1β和TNF-α的分泌,该通道可能是石英致细胞炎性反应的早期信号调节蛋白.
목적 탐토개격활갑통도(KCa)재석영치대서폐포거서세포손상중적작용.방법 대서폐포거서세포(NR8383)위수시세포;이100 μg/ml정체화무정형석영분별염진,위정체석영조화무정형석영조;불시가임하처리,정상배양적세포위음성대조조.진행체외실험.수선비교2충석영각자적작용,연후재100 μg/ml정체석영적기출상가입불동제량3충류형적KCa특이성조단제(BK조단제Paxilline、IK조단제Tram-34、SK조단제Apamin),관찰통도조단제대석영치세포손상적영향.측정세포존활솔、유산탈경매(LDH)활력、백개소-1β(IL-1β)이급종류배사인자(TNF-α)석방량.결과 여음성대조조비교,무정형석영조화정체석영조세포존활솔강저,LDH활력승고,IL-1β화TNF-α석방량증다;여무정형석영조비교,정체석영조세포존활솔명현강저,LDH활력、IL-1β、TNF-α석방량명현승고,차이균유통계학의의(P＜0.01).여정체석영조비교,IK조단제조세포존활솔증고,차이유통계학의의(P＜0.01);여정체석영조비교,BK、IK、SK조단제조세포LDH활력강저;BK、IK、SK조단제조IL-1β석방량감소,BK、IK조단제조TNF-α분비량감소,차이균유통계학의의(P＜0.05).Tram-34강저IL-1β화TNF-α석방량정제량-효응관계.결론 조단대서폐포거서세포KCa가강저석영소치세포막파배,감소염성인자IL-1β화TNF-α적분비,해통도가능시석영치세포염성반응적조기신호조절단백.
Objective To investigate the role of calcium activated-potassium channels (KCa) in the injury to rat alveolar macrophages induced by quartz.Methods The experiments were conducted on a rat alveolar macrophage cell line (NR8383) in vitro,where crystal silica (100 μg/ml) and amorphous silica (100 μg/ml) were used as the test substances and the cells without any treatment as negative controls.At first the effects of two kinds of quartzs were compared.Then KCa special inhibitors (Paxilline for BK,Tram-34 for IK,Apamin for SK) were added in different doses to the in vitro test system with 100 μg/ml crystal quartz as matrix,to observe the function of such channels.Cell viability,lactate dehydrogenase (LDH),intedeukin-1β (IL-1β) and tumor necrosis factor-a (TNF-α) were tested.Results Comparing to the negative control group,cell viability reduced,LDH leakage,IL-1β and TNF-α release increased significantly in the amorphous quartz group,furthermore,the effects by crystal quartz were much more serious than those by amorphous quartz,with a statistical significance (P＜0.01).Comparing to the crystal quartz group,IK blockers (Tram-β4) led to increase in cell viability significantly,with a statistical significance (P＜0.01); all the KCa specific blockers (Paxilline,Tram-34,Apamin) could reduce LDH leakage and IL-1 β release,with a statistical significance (P＜0.05); meanwhile,BK and IK blockers (Paxilline,Tram-34) were able to reduce TNF-α release,with a statistical significance (P＜0.05).Reduction of IL-lβ and TNF-α by Tram-34 was dose-dependent,but not so in the other two blockers.Conclusion Blocking calcium-activated potassium channels (KCa) could reduce cell membrane damage as well as IL-1β and TNF-α release induced by crystal quartz in the rat alveolar macrophages cell line in vitro,which might serve as a signal in the early regulation of inflammatory responses by quartz.