CAJ | 학술논문

目的观察不同碘营养水平时体外培养哺乳期乳腺细胞钠碘转运体(NIS)表达水平,以及肿瘤坏死因子α(TNF-α)对其的影响作用.方法体外培养小鼠原代哺乳期乳腺细胞,分为低碘1组、低碘2组、适碘组、高碘1组、高碘2组,分别加入含碘0、5、50、3000、10 000μg/L的DEME/F12培养液培养24 h,然后部分换成含对应剂量的碘加TNF-α(10-2 mg/L)的DEME/F12培养液,继续培养24 h.用实时荧光定量PCR方法检测哺乳期乳腺细胞NIS mRNA表达水平,并用In-Cell Western方法检测NIS蛋白表达水平.结果单纯加碘时低碘1组[(64.66±14.99)×10-4]乳腺细胞NIS mRNA表达水平高于适碘组[(22.76±7.36)×10-4,P＜0.05],高碘1组[(10.18±3.53)×10-4]、高碘2组[(8.59±2.89)×10-4]低于适碘组(P均＜0.05);随着碘剂量升高,乳腺细胞NIS mRNA表达水平呈下降趋势.在碘加TNF-α时,低碘1组、低碘2组、适碘组[(2.72±0.45)、(2.69±0.68)、(1.80±0.67)×10-4]乳腺细胞NIS mRNA表达水平低于单纯加碘时对应各组[(64.66±14.99)、(29.82±4.47)、(22.76±7.36)×10-4,P均＜0.05];高碘1组[(6.58±2.87)×10-4]、高碘2组[(7.04±.1.36)×10-4]高于适碘组(P均＜0.05);随着碘缺乏和碘过量程度加重,乳腺细胞NIS mRNA表达水平呈上升趋势.随着碘剂量升高,单纯加碘时乳腺细胞NIS蛋白表达水平呈下降趋势.碘加TNF-α时各组乳腺细胞NIS蛋白表达水平与单纯加碘时对应各组比较有降低的趋势.碘加TNF-α时,随着碘缺乏和碘过量程度加重,乳腺细胞NIS蛋白表达水平均呈上升趋势.结论随着碘剂量升高,哺乳期乳腺细胞NIS mRNA和蛋白表达水平均呈下降趋势.TNF-α对不同碘营养水平下哺乳期乳腺细胞NIS mRNA和蛋白表达具有抑制作用.
목적 관찰불동전영양수평시체외배양포유기유선세포납전전운체(NIS)표체수평,이급종류배사인자α(TNF-α)대기적영향작용.방법 체외배양소서원대포유기유선세포,분위저전1조、저전2조、괄전조、고전1조、고전2조,분별가입함전0、5、50、3000、10 000μg/L적DEME/F12배양액배양24 h,연후부분환성함대응제량적전가TNF-α(10-2 mg/L)적DEME/F12배양액,계속배양24 h.용실시형광정량PCR방법검측포유기유선세포NIS mRNA표체수평,병용In-Cell Western방법검측NIS단백표체수평.결과 단순가전시저전1조[(64.66±14.99)×10-4]유선세포NIS mRNA표체수평고우괄전조[(22.76±7.36)×10-4,P＜0.05],고전1조[(10.18±3.53)×10-4]、고전2조[(8.59±2.89)×10-4]저우괄전조(P균＜0.05);수착전제량승고,유선세포NIS mRNA표체수평정하강추세.재전가TNF-α시,저전1조、저전2조、괄전조[(2.72±0.45)、(2.69±0.68)、(1.80±0.67)×10-4]유선세포NIS mRNA표체수평저우단순가전시대응각조[(64.66±14.99)、(29.82±4.47)、(22.76±7.36)×10-4,P균＜0.05];고전1조[(6.58±2.87)×10-4]、고전2조[(7.04±.1.36)×10-4]고우괄전조(P균＜0.05);수착전결핍화전과량정도가중,유선세포NIS mRNA표체수평정상승추세.수착전제량승고,단순가전시유선세포NIS단백표체수평정하강추세.전가TNF-α시각조유선세포NIS단백표체수평여단순가전시대응각조비교유강저적추세.전가TNF-α시,수착전결핍화전과량정도가중,유선세포NIS단백표체수평균정상승추세.결론 수착전제량승고,포유기유선세포NIS mRNA화단백표체수평균정하강추세.TNF-α대불동전영양수평하포유기유선세포NIS mRNA화단백표체구유억제작용.
Objective To observe the expression of Na+/I- symporter(NIS) in cultured lactating mammary cells with different levels of iodine and the effect of tumor necrosis factor-α(TNF-α). Methods Original generation of mouse lactating mammary cells cultured in vitro were divided into low iodine group Ⅰ (LI-Ⅰ), low iodine group Ⅱ (LI-Ⅱ), adequate iodine group(AI), high iodine group Ⅰ(HI-Ⅰ), and high iodine group Ⅱ(HI-Ⅱ). Cells were cultured in DEME/F12 culture medium for 24 h with different concentrations of iodine (0,5,50,3000 and 10 000 μg/L, respectively), and TNF-α( 10-2 mg/L) was added to some of cultured cells for 24 h. The expression of NIS mRNA of lactating mammary cells was determined by real-time quantitative PCR and the expression of NIS protein was detected by In-Cell Western. Results In iodine alone group, the expression of NIS mRNA in LI-Ⅰ group [ (64.66 ± 14.99) x 10-4] was higher than that of AI group[ (22.76 ± 7.36) × 10-4, P ＜ 0.01 ]; HI-I group[ (10.18 ±3.53) × 10-4] and HI-Ⅱ group[ (8.59 ± 2.89) × 10-4] were lower than that of AI group(all P ＜ .0.05); With increased iodine concentration, the expression of NIS mRNA decreased. The expression of NIS mRNA in LI-Ⅰ group [(2.72 ± 0.45) × 10-4], LI-Ⅱ group[ (2.69 ± 0.68) × 10-4] and AI group[(1.80 ± 0.67) × 10-4] with iodine plus TNF-o were all lower than that of LI-Ⅰ group, LI-Ⅱ group[ (29.82 ± 4.47 ) × 10-4], and AI group without TNF-α (all P ＜ 0.01). In iodine plus TNF-α, the expression of NIS mRNA in HI-Ⅰ group[(6.58 ± 2.87) × 10-4] and HI-Ⅱ[(7.04 ± 1.36) × 10-4] group were all higher than that of AI group(all P ＜ 0.05); With increased iodine deficiency or iodine excess, the expression of NIS mRNA increased. With increased iodine concentration, the expression of NIS protein decreased in iodine alone group. The expression of NIS protein in iodine plus TNF-α was all lower than that in iodine alone group. In iodine plus TNF-α, the expression of NIS protein increased in both iodine deficiency and iodine excess conditions. Conclusions Iodine may decrease the expression of NIS mRNA and protein of lactating mammary cells. The expression of NIS mRNA and protein of lactating mammary cells was inhibited by TNF-α under different levels of iodine.