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
2008年
7期
391-394
,共4页
沈福海%范雪云%刘秉慈%贾效伟%高艾%叶萌%杜宏举%尤宝荣%史香林
瀋福海%範雪雲%劉秉慈%賈效偉%高艾%葉萌%杜宏舉%尤寶榮%史香林
침복해%범설운%류병자%가효위%고애%협맹%두굉거%우보영%사향림
石英%细胞周期蛋白质依赖激酶类%细胞周期蛋白D1%转录因子AP-1
石英%細胞週期蛋白質依賴激酶類%細胞週期蛋白D1%轉錄因子AP-1
석영%세포주기단백질의뢰격매류%세포주기단백D1%전록인자AP-1
Quartz%Cyclin-dependent kinases%Cyelin D1%Tmnscdptionfactor AP-1
目的 探讨石英暴露的人胚肺成纤维细胞(human embryonic lung fibroblast,HELF)中细胞周期蛋白D1(cyclin D1)-细胞周期依赖蛋白依赖激酶4(CDK4)蛋白的表达水平,同时探讨细胞外调节蛋白激酶(ERK)、JNK、p38和核转录因子(AP-1)等信号蛋白在石英诱导eyelin D1-CDK4蛋白表达改变中的作用.方法 石英刺激HELF后,收获细胞,检测cyclin D1和CDK4蛋白表达.选用特异化学抑制剂或分子抑制剂抑制ERK、JNK、p38或AP-1的活性后,分别采用免疫细胞化学和免疫蛋白印迹方法检测HELF中cyclin D1和CDK4蛋白表达变化.结果 HELF暴露于石英粉尘2h后,cyclin D1和CDK4蛋白表达水平分别为(7.91±0.29)x103和(5.17±0.28)x104,均明显低于HELF组,差异有统计学意义(P＜0.05).用ERK的化学抑制剂或分子抑制剂抑制ERK的活力后,能够防止石英诱导的cyclin D1和CDK4蛋白表达降低.用SP600125抑制JNK的活力后,可以防止cyclin DI和CDK4蛋白表达降低.但是抑制p38的活力对石英诱导的cyclin D1和CDK4蛋白表达降低均没有作用.用姜黄素抑制AP-1的活性后,只能防止石英诱导的CDK4的表达降低,而对cyefin D1表达降低没有影响.结论 石英诱导HELF中cyclin D1和CDK4蛋白表达降低与ERK和JNK蛋白激酶有关.AP-1与石英诱导的CDK4蛋白表达降低有关.
目的 探討石英暴露的人胚肺成纖維細胞(human embryonic lung fibroblast,HELF)中細胞週期蛋白D1(cyclin D1)-細胞週期依賴蛋白依賴激酶4(CDK4)蛋白的錶達水平,同時探討細胞外調節蛋白激酶(ERK)、JNK、p38和覈轉錄因子(AP-1)等信號蛋白在石英誘導eyelin D1-CDK4蛋白錶達改變中的作用.方法 石英刺激HELF後,收穫細胞,檢測cyclin D1和CDK4蛋白錶達.選用特異化學抑製劑或分子抑製劑抑製ERK、JNK、p38或AP-1的活性後,分彆採用免疫細胞化學和免疫蛋白印跡方法檢測HELF中cyclin D1和CDK4蛋白錶達變化.結果 HELF暴露于石英粉塵2h後,cyclin D1和CDK4蛋白錶達水平分彆為(7.91±0.29)x103和(5.17±0.28)x104,均明顯低于HELF組,差異有統計學意義(P＜0.05).用ERK的化學抑製劑或分子抑製劑抑製ERK的活力後,能夠防止石英誘導的cyclin D1和CDK4蛋白錶達降低.用SP600125抑製JNK的活力後,可以防止cyclin DI和CDK4蛋白錶達降低.但是抑製p38的活力對石英誘導的cyclin D1和CDK4蛋白錶達降低均沒有作用.用薑黃素抑製AP-1的活性後,隻能防止石英誘導的CDK4的錶達降低,而對cyefin D1錶達降低沒有影響.結論 石英誘導HELF中cyclin D1和CDK4蛋白錶達降低與ERK和JNK蛋白激酶有關.AP-1與石英誘導的CDK4蛋白錶達降低有關.
목적 탐토석영폭로적인배폐성섬유세포(human embryonic lung fibroblast,HELF)중세포주기단백D1(cyclin D1)-세포주기의뢰단백의뢰격매4(CDK4)단백적표체수평,동시탐토세포외조절단백격매(ERK)、JNK、p38화핵전록인자(AP-1)등신호단백재석영유도eyelin D1-CDK4단백표체개변중적작용.방법 석영자격HELF후,수획세포,검측cyclin D1화CDK4단백표체.선용특이화학억제제혹분자억제제억제ERK、JNK、p38혹AP-1적활성후,분별채용면역세포화학화면역단백인적방법검측HELF중cyclin D1화CDK4단백표체변화.결과 HELF폭로우석영분진2h후,cyclin D1화CDK4단백표체수평분별위(7.91±0.29)x103화(5.17±0.28)x104,균명현저우HELF조,차이유통계학의의(P＜0.05).용ERK적화학억제제혹분자억제제억제ERK적활력후,능구방지석영유도적cyclin D1화CDK4단백표체강저.용SP600125억제JNK적활력후,가이방지cyclin DI화CDK4단백표체강저.단시억제p38적활력대석영유도적cyclin D1화CDK4단백표체강저균몰유작용.용강황소억제AP-1적활성후,지능방지석영유도적CDK4적표체강저,이대cyefin D1표체강저몰유영향.결론 석영유도HELF중cyclin D1화CDK4단백표체강저여ERK화JNK단백격매유관.AP-1여석영유도적CDK4단백표체강저유관.
Objective To study the expression level of cyclin D1-CDK4 protein in human embryonic lung fthroblasts (HELF) induced by quartz, and to study whether the expression level of eyclin D1-CDK4 protein mediated by mitogen activated protein kinase (MAPK)/(AP-1 )signaling pathways. Methods Cells were harvested after stimulation 2h for the detection of cytokines. Cyclin D1 and CDK4 (cyclin dependent kinase 4)proteins was measured by immunocytochemistry(IC) and Western blot(WB). Results The exposure of HELF to crystalline quartz for 2 hours could cause the decrease of cyclin D1 and cyclin dependent kinase 4 (CDK4)protein expression level, (7.91±0.29)x 103 and (5.17±0.28 )×104 respectively, which was lower than that of the HELF group (P＜0.05). AG126 (chemical inhibitor of the extracel-lular signalregulated protein kinase (ERK) signaling pathway) and the dominant negative mutant of ERK2 (molecular inhibitor of ERK2), prevented the decrease of cyclin D1 and CDK4 protein expression level. The chemical inhibitor of c-Jun NH2-terminal amino kinase (JNK),SP600125,could prevent both eyclin D1 and CDK4 protein expression level decrease. ButSB203580, the chemical inhibitor of p38, prevented neither cyclin D1 nor CDK4 protein expression level decrease. Curcumin could prevent CDK4 protein expression level decrease but not cyclin D1 protein. ConclusionERKs and JNKs, but not p38, are responsible for the decrease of eyclin D1 and CDK4 protein expression level in HELF induced by quartz. AP-1 is responsible for the decrease of CDK4 expression level but not that of cyclin D1.
