目的 观察不同碘营养水平时胰岛素样生长因子Ⅰ(IGF-Ⅰ)和转化生长因子βⅠ(TGF-β1)对胎盘绒毛滋养层细胞(HPT-8)钠碘转运体(NIS)及pendrin mRNA表达的影响.方法 体外培养HPT-8细胞,取对数生长期细胞接种于细胞培养瓶,待细胞贴壁后根据加入培养液的不同碘含量(0、5、50、500、5000 μg/L)分为低碘1组、低碘2组、对照组、高碘1组和高碘2组.培养24h后,每组细胞再以原来的碘水平,分别加入IGF-Ⅰ(0.050 mg/L)、TGF-β1 (0.001 mg/L),即碘+IGF-Ⅰ和碘+TGF-β1.继续培养24 h后,提取细胞总RNA,反转录合成cDNA,采用实时荧光定量PCR方法检测HPT-8细胞NIS及pendrin mRNA的表达.结果 HPT-8细胞NIS mRNA的表达:在不同碘水平,单纯加碘时NIS mRNA表达组间比较差异有统计学意义(F=3.612,P＜0.01).其中低碘1组(0.44±0.21)NIS mRNA表达显著低于对照组(1.25±0.77,P＜ 0.01).在相同碘水平时,低碘1组、高碘1组NIS mRNA表达组内比较差异有统计学意义(F值分别为13.632、6.900,P均＜0.01).其中在低碘1组,碘+ IGF-Ⅰ(1.13±0.38)和碘+TGF-β1 (0.81±0.34) NIS mRNA表达高于单纯加碘(0.44±0.21,P＜ 0.01或＜0.05);在高碘1组,碘+TGF-β1 (0.62±0.30) NIS mRNA表达显著低于单纯加碘(1.23±0.91,P＜ 0.01).HPT-8细胞pendrin mRNA的表达:在不同碘水平,单纯加碘时pendrin mRNA表达组间比较差异有统计学意义(F=12.717,P＜0.01).其中低碘1组(0.59±0.15) pendrin mRNA表达显著低于对照组(1.03±0.14,P＜ 0.01),高碘1组(1.29±0.31)高于对照组(P＜0.05).在相同碘水平时,低碘1组、低碘2组、对照组、高碘1组pendrin mRNA表达组内比较差异有统计学意义(F值分别为12.588、4.588、8.679、8.445,P均＜0.01).其中在低碘1组、低碘2组和对照组,碘+IGF-Ⅰ(1.68±0.82、1.51±0.79、1.50±0.51) pendrin mRNA表达均高于单纯加碘(0.59±0.15、0.89±0.22、1.03±0.14,P均＜0.01);在高碘1组,碘+TGF-β1 (0.78±0.20) pendrin mRNA表达显著低于单纯加碘(1.29±0.31,P＜ 0.01).结论 在碘缺乏情况下,HPT-8细胞NIS、pendrin mRNA表达下降,摄碘能力下降;在轻度碘过量时,HPT-8细胞pendrin mRNA表达增加,摄碘能力增强.细胞因子IGF-Ⅰ及TGF-β1对于HPT-8细胞的碘转运能力均有一定的调节作用,即碘缺乏时增加碘的摄入,在碘过量时减少碘的摄入.
目的 觀察不同碘營養水平時胰島素樣生長因子Ⅰ(IGF-Ⅰ)和轉化生長因子βⅠ(TGF-β1)對胎盤絨毛滋養層細胞(HPT-8)鈉碘轉運體(NIS)及pendrin mRNA錶達的影響.方法 體外培養HPT-8細胞,取對數生長期細胞接種于細胞培養瓶,待細胞貼壁後根據加入培養液的不同碘含量(0、5、50、500、5000 μg/L)分為低碘1組、低碘2組、對照組、高碘1組和高碘2組.培養24h後,每組細胞再以原來的碘水平,分彆加入IGF-Ⅰ(0.050 mg/L)、TGF-β1 (0.001 mg/L),即碘+IGF-Ⅰ和碘+TGF-β1.繼續培養24 h後,提取細胞總RNA,反轉錄閤成cDNA,採用實時熒光定量PCR方法檢測HPT-8細胞NIS及pendrin mRNA的錶達.結果 HPT-8細胞NIS mRNA的錶達:在不同碘水平,單純加碘時NIS mRNA錶達組間比較差異有統計學意義(F=3.612,P＜0.01).其中低碘1組(0.44±0.21)NIS mRNA錶達顯著低于對照組(1.25±0.77,P＜ 0.01).在相同碘水平時,低碘1組、高碘1組NIS mRNA錶達組內比較差異有統計學意義(F值分彆為13.632、6.900,P均＜0.01).其中在低碘1組,碘+ IGF-Ⅰ(1.13±0.38)和碘+TGF-β1 (0.81±0.34) NIS mRNA錶達高于單純加碘(0.44±0.21,P＜ 0.01或＜0.05);在高碘1組,碘+TGF-β1 (0.62±0.30) NIS mRNA錶達顯著低于單純加碘(1.23±0.91,P＜ 0.01).HPT-8細胞pendrin mRNA的錶達:在不同碘水平,單純加碘時pendrin mRNA錶達組間比較差異有統計學意義(F=12.717,P＜0.01).其中低碘1組(0.59±0.15) pendrin mRNA錶達顯著低于對照組(1.03±0.14,P＜ 0.01),高碘1組(1.29±0.31)高于對照組(P＜0.05).在相同碘水平時,低碘1組、低碘2組、對照組、高碘1組pendrin mRNA錶達組內比較差異有統計學意義(F值分彆為12.588、4.588、8.679、8.445,P均＜0.01).其中在低碘1組、低碘2組和對照組,碘+IGF-Ⅰ(1.68±0.82、1.51±0.79、1.50±0.51) pendrin mRNA錶達均高于單純加碘(0.59±0.15、0.89±0.22、1.03±0.14,P均＜0.01);在高碘1組,碘+TGF-β1 (0.78±0.20) pendrin mRNA錶達顯著低于單純加碘(1.29±0.31,P＜ 0.01).結論 在碘缺乏情況下,HPT-8細胞NIS、pendrin mRNA錶達下降,攝碘能力下降;在輕度碘過量時,HPT-8細胞pendrin mRNA錶達增加,攝碘能力增彊.細胞因子IGF-Ⅰ及TGF-β1對于HPT-8細胞的碘轉運能力均有一定的調節作用,即碘缺乏時增加碘的攝入,在碘過量時減少碘的攝入.
목적 관찰불동전영양수평시이도소양생장인자Ⅰ(IGF-Ⅰ)화전화생장인자βⅠ(TGF-β1)대태반융모자양층세포(HPT-8)납전전운체(NIS)급pendrin mRNA표체적영향.방법 체외배양HPT-8세포,취대수생장기세포접충우세포배양병,대세포첩벽후근거가입배양액적불동전함량(0、5、50、500、5000 μg/L)분위저전1조、저전2조、대조조、고전1조화고전2조.배양24h후,매조세포재이원래적전수평,분별가입IGF-Ⅰ(0.050 mg/L)、TGF-β1 (0.001 mg/L),즉전+IGF-Ⅰ화전+TGF-β1.계속배양24 h후,제취세포총RNA,반전록합성cDNA,채용실시형광정량PCR방법검측HPT-8세포NIS급pendrin mRNA적표체.결과 HPT-8세포NIS mRNA적표체:재불동전수평,단순가전시NIS mRNA표체조간비교차이유통계학의의(F=3.612,P＜0.01).기중저전1조(0.44±0.21)NIS mRNA표체현저저우대조조(1.25±0.77,P＜ 0.01).재상동전수평시,저전1조、고전1조NIS mRNA표체조내비교차이유통계학의의(F치분별위13.632、6.900,P균＜0.01).기중재저전1조,전+ IGF-Ⅰ(1.13±0.38)화전+TGF-β1 (0.81±0.34) NIS mRNA표체고우단순가전(0.44±0.21,P＜ 0.01혹＜0.05);재고전1조,전+TGF-β1 (0.62±0.30) NIS mRNA표체현저저우단순가전(1.23±0.91,P＜ 0.01).HPT-8세포pendrin mRNA적표체:재불동전수평,단순가전시pendrin mRNA표체조간비교차이유통계학의의(F=12.717,P＜0.01).기중저전1조(0.59±0.15) pendrin mRNA표체현저저우대조조(1.03±0.14,P＜ 0.01),고전1조(1.29±0.31)고우대조조(P＜0.05).재상동전수평시,저전1조、저전2조、대조조、고전1조pendrin mRNA표체조내비교차이유통계학의의(F치분별위12.588、4.588、8.679、8.445,P균＜0.01).기중재저전1조、저전2조화대조조,전+IGF-Ⅰ(1.68±0.82、1.51±0.79、1.50±0.51) pendrin mRNA표체균고우단순가전(0.59±0.15、0.89±0.22、1.03±0.14,P균＜0.01);재고전1조,전+TGF-β1 (0.78±0.20) pendrin mRNA표체현저저우단순가전(1.29±0.31,P＜ 0.01).결론 재전결핍정황하,HPT-8세포NIS、pendrin mRNA표체하강,섭전능력하강;재경도전과량시,HPT-8세포pendrin mRNA표체증가,섭전능력증강.세포인자IGF-Ⅰ급TGF-β1대우HPT-8세포적전전운능력균유일정적조절작용,즉전결핍시증가전적섭입,재전과량시감소전적섭입.
Objective To observe the effects of insulin-like growth factor-Ⅰ (IGF-Ⅰ) and transforming growth factor-β1 (TGF-β1) on the expressions of sodium iodide symporter(NIS) and pendrin mRNA in a placental villous trophoblast cell line(HPT-8) exposed to different levels of iodine.Methods HPT-8 cells were cultured in vitro in the culture flask and divided into low iodine group-Ⅰ (LI-Ⅰ),low iodine group-Ⅱ (LI-Ⅱ),control group,high iodine group-Ⅰ (HI-Ⅰ) and high iodine group-Ⅱ (HI-Ⅱ) that exposed to different concentrations of iodine (0,5,50,500,5000 μg/L).After cell cultured for 24 h,the followings were added to the culture medium:iodine plus IGF-Ⅰ(0.050 mg/L),iodine plus TGF-β1 (0.001 mg/L).After cultured for another 24 h,total RNA was extracted,the expressions of NIS and pendrin mRNA of HPT-8 cells were determined by real-time quantitative PCR.Results The expression of NIS mRNA in HPT-8 cells:at different levels of iodine,the differences of NIS mRNA expression between groups were statistically significant in group with iodine alone(F =3.612,P ＜ 0.01).The expression of NIS mRNA in LI-Ⅰ group(0.44 ± 0.21) was significantly lower than that of control group(1.25 ± 0.77,P＜ 0.01).At the same level of iodine,in LI-Ⅰ group and HI-Ⅰ group,the differences of NIS mRNA expression within groups were statistically significant (F =13.632,6.900,all P ＜ 0.01).In LI-Ⅰ group,the expressions of NIS mRNA were higher in iodine plus IGF-Ⅰ(1.13 ± 0.38) and iodine plus TGF-β1 (0.81 ± 0.34) than that of pure iodine(0.44 ± 0.21,P ＜ 0.01 or ＜ 0.05);in HI-Ⅰ group,the expression of NIS mRNA was lower in iodine plus TGF-β1 (0.62 ± 0.30) than that of pure iodine(1.23 ± 0.91,P ＜ 0.01).The expression of pendrin mRNA in HPT-8 cells:at different levels of iodine,the differences of pendrin mRNA expression between groups were statistically significant in group with iodine alone(F =12.717,P ＜ 0.01).The expression of pendrin mRNA in LI-Ⅰ group(0.59 ± 0.15) was significantly lower than that of control group(1.03 ± 0.14,P ＜ 0.01) ; HI-Ⅰ group(1.29 ± 0.31) was higher than control group(P ＜ 0.05).At the same level of iodine,the differences of pendrin mRNA expression within groups were statistically significant in LI-Ⅰ,LI-Ⅱ,control and HI-Ⅰ groups (F=12.588,4.588,8.679,8.445,all P ＜ 0.01).In LI-Ⅰ,LI-Ⅱ and control groups,the expressions of pendrin mRNA were significantly higher in iodine plus IGF-Ⅰ(1.68 ± 0.82,1.51 ± 0.79,1.50 ± 0.51) than that of pure iodine(0.59 ± 0.15,0.89 ± 0.22,1.03 ± 0.14,all P ＜ 0.01); in HI-Ⅰ group,the expression of pendrin mRNA was significantly lower in iodine plus TGF-β1 (0.78 ± 0.20) than that of pure iodine(1.29 ± 0.31,P ＜ 0.01).Conclusions In the case of iodine deficiency,the mRNA expressions of NIS and pendrin in HPT-8 cells are decreased and the iodine uptake ability is decreased; the expression of pendrin mRNA in HPT-8 cells is increased and placental iodine uptake is increased under the conditions of mild iodine excessive.IGF-Ⅰ and TGF-β1 play a role in the placental iodine uptake through increasing iodine uptake under the conditions of iodine deficiency and decreasing iodine uptake under the conditions of iodine excessive.
