中国临床康复
中國臨床康複
중국림상강복
CHINESE JOURNAL OF CLINICAL REHABILITATION
2006年
24期
187-189
,共3页
沈慧%秦海宏%龙建纲%王福俤%郭俊生
瀋慧%秦海宏%龍建綱%王福俤%郭俊生
침혜%진해굉%룡건강%왕복제%곽준생
锌/代谢%载体蛋白质类%小肠
鋅/代謝%載體蛋白質類%小腸
자/대사%재체단백질류%소장
背景:小肠锌吸收降低会导致皮炎、脱发、生长发育障碍等.低锌状态下如何保持小肠锌内稳态的作用途径至今尚不清楚,锌转运体的发现及相关研究为其提供了新的研究方向.目的:观察低锌浓度对二价金属离子转运体1和锌铁调控蛋白4mRNA表达的影响,分析低锌状态下小肠锌吸收的可能途径.设计:空白对照观察.单位:解放军第二军医大学海医系军队卫生学教研室.材料:实验于2004-10/2005-05在解放军第二军医大学军队卫生学教研室完成,人结肠腺癌细胞系Caco2购自上海中科院细胞所.方法:将Caco2细胞培养至一定浓度.①时间效应:应用反转录聚合酶链反应分别检测10 μmol/L TPEN暴露时,0,2,4,6,8和10 h时相点Caco2细胞二价金属离子转运体1和锌铁调控蛋白4mRNA的表达情况.②剂量效应:应用反转录聚合酶链反应分别检测0,2.5,5,7.5,10 μmol/LTPEN暴露时,各组Caco2细胞二价金属离子转运体1和锌铁调控蛋白4mRNA的表达情况.主要观察指标:锌对Caco2细胞二价金属离子转运体1和锌铁调控蛋白4mRNA表达影响的时间和剂量效应.结果:①时间效应:与0 h比较,2 h和4 h时二价金属离子转运体1mRNA表达水平没有明显变化,6 h,8 h和10 h,二价金属离子转运体1mRNA均有显著升高(P＜0.05).锌铁调控蛋白4mRNA表达水平随着时间的延长而升高,6 h达到峰值,为0 h的2.1倍.②剂量效应:二价金属离子转运体1mRNA的表达量在TPEN浓度为2.5和5 μmol/L时没有明显改变,7.5和10μmol/L时二价金属离子转运体1mRNA的表达量较对照组有显著升高(P＜0.05).锌铁调控蛋白4mRNA的表达量随着TPEN浓度的升高而升高,10 μmol/L时锌铁调控蛋白4mRNA表达量为0μmol/L时的2.7倍.结论:Caco2细胞二价金属离子转运体1和锌铁调控蛋白4mRNA表达受锌的调控,并且有时间和剂量效应,但是锌铁调控蛋白4mRNA变化较二价金属离子转运体1mRNA变化敏感且迅速.低锌状态下,细胞可能通过上调二价金属离子转运体1和锌铁调控蛋白4mRNA表达水平而调节细胞锌内稳态.
揹景:小腸鋅吸收降低會導緻皮炎、脫髮、生長髮育障礙等.低鋅狀態下如何保持小腸鋅內穩態的作用途徑至今尚不清楚,鋅轉運體的髮現及相關研究為其提供瞭新的研究方嚮.目的:觀察低鋅濃度對二價金屬離子轉運體1和鋅鐵調控蛋白4mRNA錶達的影響,分析低鋅狀態下小腸鋅吸收的可能途徑.設計:空白對照觀察.單位:解放軍第二軍醫大學海醫繫軍隊衛生學教研室.材料:實驗于2004-10/2005-05在解放軍第二軍醫大學軍隊衛生學教研室完成,人結腸腺癌細胞繫Caco2購自上海中科院細胞所.方法:將Caco2細胞培養至一定濃度.①時間效應:應用反轉錄聚閤酶鏈反應分彆檢測10 μmol/L TPEN暴露時,0,2,4,6,8和10 h時相點Caco2細胞二價金屬離子轉運體1和鋅鐵調控蛋白4mRNA的錶達情況.②劑量效應:應用反轉錄聚閤酶鏈反應分彆檢測0,2.5,5,7.5,10 μmol/LTPEN暴露時,各組Caco2細胞二價金屬離子轉運體1和鋅鐵調控蛋白4mRNA的錶達情況.主要觀察指標:鋅對Caco2細胞二價金屬離子轉運體1和鋅鐵調控蛋白4mRNA錶達影響的時間和劑量效應.結果:①時間效應:與0 h比較,2 h和4 h時二價金屬離子轉運體1mRNA錶達水平沒有明顯變化,6 h,8 h和10 h,二價金屬離子轉運體1mRNA均有顯著升高(P＜0.05).鋅鐵調控蛋白4mRNA錶達水平隨著時間的延長而升高,6 h達到峰值,為0 h的2.1倍.②劑量效應:二價金屬離子轉運體1mRNA的錶達量在TPEN濃度為2.5和5 μmol/L時沒有明顯改變,7.5和10μmol/L時二價金屬離子轉運體1mRNA的錶達量較對照組有顯著升高(P＜0.05).鋅鐵調控蛋白4mRNA的錶達量隨著TPEN濃度的升高而升高,10 μmol/L時鋅鐵調控蛋白4mRNA錶達量為0μmol/L時的2.7倍.結論:Caco2細胞二價金屬離子轉運體1和鋅鐵調控蛋白4mRNA錶達受鋅的調控,併且有時間和劑量效應,但是鋅鐵調控蛋白4mRNA變化較二價金屬離子轉運體1mRNA變化敏感且迅速.低鋅狀態下,細胞可能通過上調二價金屬離子轉運體1和鋅鐵調控蛋白4mRNA錶達水平而調節細胞鋅內穩態.
배경:소장자흡수강저회도치피염、탈발、생장발육장애등.저자상태하여하보지소장자내은태적작용도경지금상불청초,자전운체적발현급상관연구위기제공료신적연구방향.목적:관찰저자농도대이개금속리자전운체1화자철조공단백4mRNA표체적영향,분석저자상태하소장자흡수적가능도경.설계:공백대조관찰.단위:해방군제이군의대학해의계군대위생학교연실.재료:실험우2004-10/2005-05재해방군제이군의대학군대위생학교연실완성,인결장선암세포계Caco2구자상해중과원세포소.방법:장Caco2세포배양지일정농도.①시간효응:응용반전록취합매련반응분별검측10 μmol/L TPEN폭로시,0,2,4,6,8화10 h시상점Caco2세포이개금속리자전운체1화자철조공단백4mRNA적표체정황.②제량효응:응용반전록취합매련반응분별검측0,2.5,5,7.5,10 μmol/LTPEN폭로시,각조Caco2세포이개금속리자전운체1화자철조공단백4mRNA적표체정황.주요관찰지표:자대Caco2세포이개금속리자전운체1화자철조공단백4mRNA표체영향적시간화제량효응.결과:①시간효응:여0 h비교,2 h화4 h시이개금속리자전운체1mRNA표체수평몰유명현변화,6 h,8 h화10 h,이개금속리자전운체1mRNA균유현저승고(P＜0.05).자철조공단백4mRNA표체수평수착시간적연장이승고,6 h체도봉치,위0 h적2.1배.②제량효응:이개금속리자전운체1mRNA적표체량재TPEN농도위2.5화5 μmol/L시몰유명현개변,7.5화10μmol/L시이개금속리자전운체1mRNA적표체량교대조조유현저승고(P＜0.05).자철조공단백4mRNA적표체량수착TPEN농도적승고이승고,10 μmol/L시자철조공단백4mRNA표체량위0μmol/L시적2.7배.결론:Caco2세포이개금속리자전운체1화자철조공단백4mRNA표체수자적조공,병차유시간화제량효응,단시자철조공단백4mRNA변화교이개금속리자전운체1mRNA변화민감차신속.저자상태하,세포가능통과상조이개금속리자전운체1화자철조공단백4mRNA표체수평이조절세포자내은태.
BACKGROUND: Decreasing of absorption of zinc from small intestine can induce dermatitis, alopecia, growth and developmental disorder and so on. It is not very clear that how to keep homeostasis when there is low zinc concentration. The discoveries of zinc transporters and relative researches provide new study direction.OBJECTIVE: To study the effect of different zinc concentration on the expressions of divalent metal transporter 1 (DMT1) and human zinc-regulated transporter (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) 4mRNA, and analyze the potential pathway of absorption of zinc from small intestine in low zinc concentration.DESIGN: Blank-controlled experiment.SETTING: Department of Military Hygiene, Navy Faculty, Second Military Medical University of Chinese PLA.MATERIALS: The experiment was accomplished in Department of Military Hygiene, Navy Faculty of Second Military Medical University of Chinese PLA between October 2004 and May 2005. Materials were human colon adenocarcinoma cells Caco2 that were purchased from Shanghai Institute of Cell of Chinese Academy of Sciences.Time-dependent effect: The expression of DMT1 and ZIP4 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR) at 0,dependent effect: The expression of DMT1 and ZIP4 mRNA were measured after the 0, 2.5, 5, 7.5 and 10 μmol/L TPEN exposure, respectively, by RT-PCR.MAIN OUTCOME MEASURES: Time-and dose-dependent effect of zinc on the expression of DMT1 and ZIP4 mRNA in Caco2 cells.RESULTS: ①Time-dependent effect: Compared with 0 hour, the expression of DMT1 mRNA obviously increased at 6, 8 and 10 hours (P ＜ 0.05 ), but there was no significant change at 2 and 4 hours. The expression of ZIP4 mRNA markedly increased in all timing, and ZIP4 mRNA level at the 6th hour reached the peak with the prolongation of low zinc duration,which was 2.1 times ofthat at 0 hour. ②Dose-dependent effect: The expression of DMT1 mRNA distinctly increased at the concentration of 7.5 and 10 μmol/L TPEN exposure as compared with that of the control group (P ＜ 0.05), but there was no significant change at 2.5 and 5 μmol/L. The ZIP4 mRNA expression increased with the increasing of the concentration of TPEN, the expression of ZIP4 mRNA was 2.7 times of that at 0 μmol/L. CONCLUSION: Zinc can regulate the expression of DMT1 and ZIP4 mRNA in Caco2 cells, and there is time-and dose-dependent effect. But the mRNA expression of ZIP4 is more sensitive and prompt than DMT1. Cells can upregulate the expression of DMT1 and ZIP4 mRNA to keep the homeostasis in low zinc condition.
