中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2013年
2期
83-87
,共5页
周庆红%陈曦%冷玲%张静姝%汤乃军
週慶紅%陳晞%冷玲%張靜姝%湯迺軍
주경홍%진희%랭령%장정주%탕내군
邻苯二甲酸二丁酯%邻苯二甲酸单丁酯%小鼠睾丸间质瘤细胞%睾酮%胰岛素样因子3
鄰苯二甲痠二丁酯%鄰苯二甲痠單丁酯%小鼠睪汍間質瘤細胞%睪酮%胰島素樣因子3
린분이갑산이정지%린분이갑산단정지%소서고환간질류세포%고동%이도소양인자3
Dibutyl phthalate%Monobutyl phthalate%Leydig tumor cells of mice%Testosterone%Insulin-like factor 3
目的 观察邻苯二甲酸二丁酯(dibutyl phthalate,DBP)、邻苯二甲酸单丁酯(monobutyl phthalate,MBP)对小鼠睾丸间质瘤细胞(MA-10)中胰岛素样因子3(insulin-like factor 3,INSL3) mRNA 及蛋白的表达量和细胞睾酮分泌水平的影响.方法 以小鼠睾丸间质瘤细胞(MA-10)为模型,经过DBP、MBP直接染毒后,采用酶联免疫吸附法(ELISA)测定细胞培养基上清中睾酮的含量,采用荧光定量PCR测定细胞中INSL3 mRNA的表达,采用蛋白免疫印迹法(western blot)测定INSL3蛋白的表达水平.结果 与对照组比较,MA-10细胞睾酮合成量在低染毒剂量(10-9～10-6 mol/L)时均表现出刺激效应,睾酮含量增加；在高染毒剂量(10-3 mol/L)时表现抑制效应,随着染毒时间的延长,睾酮水平逐渐降低.荧光定量PCR以及Western blot测定结果显示,DBP在10-6、10-4 mol/L染毒时,INSL3 mRNA及蛋白表达水平较对照组降低,差异有统计学意义(P＜0.05)；随着MBP染毒剂量的增加,INSL3 mRNA及蛋白表达水平呈现下降趋势,但MBP在10-7 mol/L时,与对照组相比,对MA-10细胞中INSL3蛋白表达表现出刺激作用.结论 DBP、MBP染毒对MA-10细胞睾酮分泌表现出高剂量抑制、低剂量刺激的双向效应；DBP、MBP染毒可抑制MA-10细胞中INSL3 mRNA和蛋白的表达.
目的 觀察鄰苯二甲痠二丁酯(dibutyl phthalate,DBP)、鄰苯二甲痠單丁酯(monobutyl phthalate,MBP)對小鼠睪汍間質瘤細胞(MA-10)中胰島素樣因子3(insulin-like factor 3,INSL3) mRNA 及蛋白的錶達量和細胞睪酮分泌水平的影響.方法 以小鼠睪汍間質瘤細胞(MA-10)為模型,經過DBP、MBP直接染毒後,採用酶聯免疫吸附法(ELISA)測定細胞培養基上清中睪酮的含量,採用熒光定量PCR測定細胞中INSL3 mRNA的錶達,採用蛋白免疫印跡法(western blot)測定INSL3蛋白的錶達水平.結果 與對照組比較,MA-10細胞睪酮閤成量在低染毒劑量(10-9～10-6 mol/L)時均錶現齣刺激效應,睪酮含量增加；在高染毒劑量(10-3 mol/L)時錶現抑製效應,隨著染毒時間的延長,睪酮水平逐漸降低.熒光定量PCR以及Western blot測定結果顯示,DBP在10-6、10-4 mol/L染毒時,INSL3 mRNA及蛋白錶達水平較對照組降低,差異有統計學意義(P＜0.05)；隨著MBP染毒劑量的增加,INSL3 mRNA及蛋白錶達水平呈現下降趨勢,但MBP在10-7 mol/L時,與對照組相比,對MA-10細胞中INSL3蛋白錶達錶現齣刺激作用.結論 DBP、MBP染毒對MA-10細胞睪酮分泌錶現齣高劑量抑製、低劑量刺激的雙嚮效應；DBP、MBP染毒可抑製MA-10細胞中INSL3 mRNA和蛋白的錶達.
목적 관찰린분이갑산이정지(dibutyl phthalate,DBP)、린분이갑산단정지(monobutyl phthalate,MBP)대소서고환간질류세포(MA-10)중이도소양인자3(insulin-like factor 3,INSL3) mRNA 급단백적표체량화세포고동분비수평적영향.방법 이소서고환간질류세포(MA-10)위모형,경과DBP、MBP직접염독후,채용매련면역흡부법(ELISA)측정세포배양기상청중고동적함량,채용형광정량PCR측정세포중INSL3 mRNA적표체,채용단백면역인적법(western blot)측정INSL3단백적표체수평.결과 여대조조비교,MA-10세포고동합성량재저염독제량(10-9～10-6 mol/L)시균표현출자격효응,고동함량증가；재고염독제량(10-3 mol/L)시표현억제효응,수착염독시간적연장,고동수평축점강저.형광정량PCR이급Western blot측정결과현시,DBP재10-6、10-4 mol/L염독시,INSL3 mRNA급단백표체수평교대조조강저,차이유통계학의의(P＜0.05)；수착MBP염독제량적증가,INSL3 mRNA급단백표체수평정현하강추세,단MBP재10-7 mol/L시,여대조조상비,대MA-10세포중INSL3단백표체표현출자격작용.결론 DBP、MBP염독대MA-10세포고동분비표현출고제량억제、저제량자격적쌍향효응；DBP、MBP염독가억제MA-10세포중INSL3 mRNA화단백적표체.
Objective To observe the effects ofdibutyl phthalate (DBP) and monobutyl phthalate (MBP) on the mRNA and protein expression of insulin-like factor 3 (INSL3) in the Leydig tumor cells (MA-10) of mice and the level of testosterone secreted from MA-10 cells.Methods The MA-10 cells of mice,used as a cellular model,were exposed to DBP and MBP.The content of testosterone in the supernatant medium was measured by enzyme-linked immunosorbent assay; the mRNA and protein expression levels of INSL3 in MA-10 cells were measured by quantitative PCR and Western Blot.Results Compared with the control group,MA-10 cells showed increased synthesis of testosterone when exposed to low concentrations of DBP and MBP (10-9～10-6 mol/L)and inhibited synthesis of testosterone when exposed to high concentrations of DBP and MBP (l0-3 mol/L),and the typical two-way effects became more significant as the time went one and the concentrations increased (P＜0.05).Compared with the control group,MA-10 cells showed significantly lower mRNA and protein expression levels of INSL3 when exposed to 10-6 and 10-4 mol/L DBP (P＜0.05); MA-10 cells showed increased protein expression of INSL3 when exposed to 10-7 mol/L MBP,and the mRNA and protein expression levels of INSL3 decreased as the concentration of MBP increased.Conclusion DBP and MBP can inhibit the secretion of testosterone from MA-10 cells at high concentrations,but stimulate the secretion of testosterone at low concentrations.Both DBP and MBP have inhibitory effects on the mRNA and protein expression of INSL3 in MA-10 cells.
