CAJ | 학술논문

目的探讨煤焦沥青烟提取物(coal tar pitch smoke extract,CTP)刺激人支气管上皮细胞(BEAS-2B)是否发生炎性凋亡(pyroptosis).方法用l、3μg/ml的CTP分别染毒BEAS-2B细胞8、24 h,以二甲基亚砜(DMSO)为对照,用乳酸脱氢酶(lactic dehydrogenase,LDH)活力测定试剂盒和白细胞介素-1β(Interleukin-1 beta,IL-1β) ELISA试剂盒分别检测细胞培养上清中的LDH活力和IL-1β含量,用半胱氨酸天冬氨酸特异性蛋白酶-1(caspase-1)活力测定试剂盒检测细胞caspase-1活力.结果染毒8h,1、3 μg/ml CTP染毒组细胞中caspase-1的活力分别为(9.29±0.30) μmol/ml和(8.67±0.59) μmol/ml,均明显高于DMSO对照组[(7.42±0.59) μmol/ml],差异有统计学意义(P＜0.05)；1、3μg/mlCTP染毒组LDH活力和IL-1β含量与DMSO对照组的差异无统计学意义(P＞0.05).染毒24 h时,1、3μg/ml染毒组与DMSO对照组细胞中caspase-1活力差异无统计学意义(P＞0.05)；1、3μg/ml染毒组培养液上清中LDH活力[(1323.03±28.53)、(1148.45±16.42) U/dl]和IL-1β含量[(125.37±25.00)、(92.04±19.09) pg/m]均高于对照组[LDH:(1091.93±26.64) U/dl,IL-1β:(46.20±14.43) pg/ml],差异有统计学意义(P＜0.05).结论煤焦沥青烟提取物刺激BEAS-2B细胞早期caspase-1活力升高,继之LDH酶活力及IL-1β含量均增高,可能发生pyroptosis.
목적 탐토매초력청연제취물(coal tar pitch smoke extract,CTP)자격인지기관상피세포(BEAS-2B)시부발생염성조망(pyroptosis).방법 용l、3μg/ml적CTP분별염독BEAS-2B세포8、24 h,이이갑기아풍(DMSO)위대조,용유산탈경매(lactic dehydrogenase,LDH)활력측정시제합화백세포개소-1β(Interleukin-1 beta,IL-1β) ELISA시제합분별검측세포배양상청중적LDH활력화IL-1β함량,용반광안산천동안산특이성단백매-1(caspase-1)활력측정시제합검측세포caspase-1활력.결과 염독8h,1、3 μg/ml CTP염독조세포중caspase-1적활력분별위(9.29±0.30) μmol/ml화(8.67±0.59) μmol/ml,균명현고우DMSO대조조[(7.42±0.59) μmol/ml],차이유통계학의의(P＜0.05)；1、3μg/mlCTP염독조LDH활력화IL-1β함량여DMSO대조조적차이무통계학의의(P＞0.05).염독24 h시,1、3μg/ml염독조여DMSO대조조세포중caspase-1활력차이무통계학의의(P＞0.05)；1、3μg/ml염독조배양액상청중LDH활력[(1323.03±28.53)、(1148.45±16.42) U/dl]화IL-1β함량[(125.37±25.00)、(92.04±19.09) pg/m]균고우대조조[LDH:(1091.93±26.64) U/dl,IL-1β:(46.20±14.43) pg/ml],차이유통계학의의(P＜0.05).결론 매초력청연제취물자격BEAS-2B세포조기caspase-1활력승고,계지LDH매활력급IL-1β함량균증고,가능발생pyroptosis.
Objective To explore whether coal tar pitch smoke extract (CTP) induced pyroptosis in human bronchial epithelial cells (BEAS-2B).Methods BEAS-2B cells were treated with different concentrations of CTP (1,3 μg/ml) for 8 h and 24 h,respectively.Lactic dehydrogenase (LDH) activity and interleukin-1 beta (IL-1β) levels in the supernatants of cell culture media were measured with LDH activity or human IL-1β ELISA kit,respectively.The activity of Caspase-1 was measured with Caspase-1 colorimetric assay kit.Results The activity of caspase-1 in 1 and 3 μg/ml CTP groups were (9.29±0.30) and (8.67±0.59)μmol/ml respectivelywhich wereboth significantly increased compared to that (7.42±0.59) μmol/ml in the control group (P＜0.05) after 8 h exposure,but there was no significant difference in the activity of LDH and levels of IL-1β in the cell culture media among the CTP and control groups.24 h after exposure,the activity of LDH in the CTP (1,3 μg/ml) groups were (1323.03±28.53) and (1148.45±16.42) U/dl respectively which were significantly higher than that (1091.93±26.64) U/dl in the control group (P＜0.05),and the levels of IL-1β in the CTP (1 and 3 μg/ml) groups were (125.37±25.00) pg/ml and (92.04±19.09) pg/ml respectively which were significantly higher than that (46.20±14.43) pg/ml in the control group (P＜0.05),but there was no significant difference in the activity of Caspase-1 among CTP and control groups (P＜0.05).Conclusion CTP treatment induced early increase in caspase-1 activity followed by the increase in LDH activity and IL-1 levels,indicative of pyroptosis in human bronchial epithelial cells.