中国农业科学
中國農業科學
중국농업과학
SCIENTIA AGRICULTURA SINICA
2015年
8期
1609-1615
,共7页
王小城%熊霞%杨焕胜%高巍%龚敏%印遇龙
王小城%熊霞%楊煥勝%高巍%龔敏%印遇龍
왕소성%웅하%양환성%고외%공민%인우룡
腐胺%脯氨酸%空肠%多胺%Wnt
腐胺%脯氨痠%空腸%多胺%Wnt
부알%포안산%공장%다알%Wnt
putrescine%proline%jejunum%polyamine%Wnt
【目的】研究腐胺和脯氨酸对哺乳期仔猪空肠绒毛-隐窝轴上皮细胞的多胺（腐胺、亚精胺和精胺）代谢、Wnt信号通路关键基因的mRNA相对表达量的影响。【方法】选取18头0日龄的刚出生的三元杂交（长白×大白×杜洛克）仔猪，随机配对分成3组，每组6个重复，每个重复1头猪，对照组，腐胺组和脯氨酸组，分别灌喂等体积的生理盐水，5 mg·kg-1体重添加剂量的腐胺和25 mg·kg-1体重添加剂量的脯氨酸，到14日龄断奶，断奶后3 d屠宰，分离空肠绒毛-隐窝轴的3个不同分化程度（绒毛顶端，绒毛中段和隐窝）的细胞，分别为F1，F2， F3。高效液相色谱法测定F1，F2，F3中的多胺浓度，RT-PCR测定多胺代谢途径中的相关基因以及Wnt信号通路中关键基因的mRNA相对表达量。【结果】1、在绒毛顶端细胞F1中，与对照组相比，脯氨酸组的腐胺、亚精胺、精胺的浓度均显著增加（P＜0.05），而腐胺组的多胺均无显著差异（P＞0.05）；在绒毛中段细胞F2中，除了腐胺的含量无显著差异外，脯氨酸组和腐胺组的亚精胺、精胺的浓度均显著高于对照组（P＜0.05），且脯氨酸组的亚精胺和精胺的浓度均显著高于腐胺组（P＜0.05）；在隐窝底端细胞F3中，腐胺组和脯氨酸组的腐胺、亚精胺、精胺浓度与对照组的浓度差异均不显著（P＞0.05）。2、鸟氨酸脱羧酶(ODC)基因在绒毛顶端上皮细胞 F1中的 mRNA表达量，腐胺组显著高于脯氨酸组和对照组（P＜0.05）；精氨酸酶（arginase）在F2中的mRNA表达量，脯氨酸组显著高于腐胺组和对照组（P＜0.05）；sFRP3在绒毛中段上皮细胞F2中的mRNA表达量，脯氨酸组的显著高于腐胺组（P＞0.05）；sFRP4在隐窝上皮细胞F3中的mRNA表达量，腐胺组显著高于对照组和脯氨酸组（P＜0.05）。【结论】添加了外源腐胺和脯氨酸，促进了哺乳期仔猪的空肠绒毛-隐窝轴上皮细胞的分化细胞的多胺浓度及促进了多胺的代谢，而对未分化的隐窝细胞没显著影响，同时通过Wnt信号通路调控空肠绒毛-隐窝轴细胞的分化成熟。
【目的】研究腐胺和脯氨痠對哺乳期仔豬空腸絨毛-隱窩軸上皮細胞的多胺（腐胺、亞精胺和精胺）代謝、Wnt信號通路關鍵基因的mRNA相對錶達量的影響。【方法】選取18頭0日齡的剛齣生的三元雜交（長白×大白×杜洛剋）仔豬，隨機配對分成3組，每組6箇重複，每箇重複1頭豬，對照組，腐胺組和脯氨痠組，分彆灌餵等體積的生理鹽水，5 mg·kg-1體重添加劑量的腐胺和25 mg·kg-1體重添加劑量的脯氨痠，到14日齡斷奶，斷奶後3 d屠宰，分離空腸絨毛-隱窩軸的3箇不同分化程度（絨毛頂耑，絨毛中段和隱窩）的細胞，分彆為F1，F2， F3。高效液相色譜法測定F1，F2，F3中的多胺濃度，RT-PCR測定多胺代謝途徑中的相關基因以及Wnt信號通路中關鍵基因的mRNA相對錶達量。【結果】1、在絨毛頂耑細胞F1中，與對照組相比，脯氨痠組的腐胺、亞精胺、精胺的濃度均顯著增加（P＜0.05），而腐胺組的多胺均無顯著差異（P＞0.05）；在絨毛中段細胞F2中，除瞭腐胺的含量無顯著差異外，脯氨痠組和腐胺組的亞精胺、精胺的濃度均顯著高于對照組（P＜0.05），且脯氨痠組的亞精胺和精胺的濃度均顯著高于腐胺組（P＜0.05）；在隱窩底耑細胞F3中，腐胺組和脯氨痠組的腐胺、亞精胺、精胺濃度與對照組的濃度差異均不顯著（P＞0.05）。2、鳥氨痠脫羧酶(ODC)基因在絨毛頂耑上皮細胞 F1中的 mRNA錶達量，腐胺組顯著高于脯氨痠組和對照組（P＜0.05）；精氨痠酶（arginase）在F2中的mRNA錶達量，脯氨痠組顯著高于腐胺組和對照組（P＜0.05）；sFRP3在絨毛中段上皮細胞F2中的mRNA錶達量，脯氨痠組的顯著高于腐胺組（P＞0.05）；sFRP4在隱窩上皮細胞F3中的mRNA錶達量，腐胺組顯著高于對照組和脯氨痠組（P＜0.05）。【結論】添加瞭外源腐胺和脯氨痠，促進瞭哺乳期仔豬的空腸絨毛-隱窩軸上皮細胞的分化細胞的多胺濃度及促進瞭多胺的代謝，而對未分化的隱窩細胞沒顯著影響，同時通過Wnt信號通路調控空腸絨毛-隱窩軸細胞的分化成熟。
【목적】연구부알화포안산대포유기자저공장융모-은와축상피세포적다알（부알、아정알화정알）대사、Wnt신호통로관건기인적mRNA상대표체량적영향。【방법】선취18두0일령적강출생적삼원잡교（장백×대백×두락극）자저，수궤배대분성3조，매조6개중복，매개중복1두저，대조조，부알조화포안산조，분별관위등체적적생리염수，5 mg·kg-1체중첨가제량적부알화25 mg·kg-1체중첨가제량적포안산，도14일령단내，단내후3 d도재，분리공장융모-은와축적3개불동분화정도（융모정단，융모중단화은와）적세포，분별위F1，F2， F3。고효액상색보법측정F1，F2，F3중적다알농도，RT-PCR측정다알대사도경중적상관기인이급Wnt신호통로중관건기인적mRNA상대표체량。【결과】1、재융모정단세포F1중，여대조조상비，포안산조적부알、아정알、정알적농도균현저증가（P＜0.05），이부알조적다알균무현저차이（P＞0.05）；재융모중단세포F2중，제료부알적함량무현저차이외，포안산조화부알조적아정알、정알적농도균현저고우대조조（P＜0.05），차포안산조적아정알화정알적농도균현저고우부알조（P＜0.05）；재은와저단세포F3중，부알조화포안산조적부알、아정알、정알농도여대조조적농도차이균불현저（P＞0.05）。2、조안산탈최매(ODC)기인재융모정단상피세포 F1중적 mRNA표체량，부알조현저고우포안산조화대조조（P＜0.05）；정안산매（arginase）재F2중적mRNA표체량，포안산조현저고우부알조화대조조（P＜0.05）；sFRP3재융모중단상피세포F2중적mRNA표체량，포안산조적현저고우부알조（P＞0.05）；sFRP4재은와상피세포F3중적mRNA표체량，부알조현저고우대조조화포안산조（P＜0.05）。【결론】첨가료외원부알화포안산，촉진료포유기자저적공장융모-은와축상피세포적분화세포적다알농도급촉진료다알적대사，이대미분화적은와세포몰현저영향，동시통과Wnt신호통로조공공장융모-은와축세포적분화성숙。
[Objective]The aim of this paper is to study the effects of putrescine and proline on polyamines (putrescine, spermidine and spermine) metabolites, relative mRNA expression levels of Wnt signaling molecules along the villus-crypt axis in the jejunum of sucking piglet.[Method]Eighteen 0-day old newborn crossbred (Landrace × Yorkshire × Duroc) piglets were randomly divided into three groups:control group, putrescine group and proline group. The piglets in control group were administered an equal volume of saline, and that in other two groups received putrescine (5 mg·kg-1 body weight ) and proline (25 mg·kg-1 body weight ), respectively. Piglets were weaned at 14-day old and slaughtered on 3 days postweaning. The jejuna epithelial cells along the crypt-villus axis were isolated by Krebs Henseleit (KH) solution and yield 3“cell fractions”(Fraction 1, 2 and 3). The concentration of polyamine in F1, F2 and F3 was determined by high performance liquid chromatography, and relative mRNA expression of related genes of polyamine metabolic and wnt signaling pathways were measured by real time PCR. [Result] The concentration of putrescine, spermine and spermidine in F1 were significantly increased (P<0.05) in proline group compared to control group, while there was no significant difference in putrescine group (P>0.05). The concentrations of spermine and spermidine in F2 of proline or putrescine groups were significantly higher than that of control group (P<0.05). the concentrations of putrescine, spermine and spermidine in F3 of three group were not changed significantly (P>0.05). The expression of ornithine decarboxylase (ODC) gene in F1 of putrescine group was significantly higher than that of proline group and control group (P<0.05). The expression of arginase in F2 of proline group was significantly higher than that of putrescine group and control group (P<0.05). The expression of SFRP3 in F2 of proline group was significantly higher than that of putrescine group (P<0.05);the expression of sFRP4 in F3 of putrescine group is significantly higher than that of control group and proline group (P<0.05). [Conclusion] Exogenous putrescine and proline increased the concentration of polyamine in differentiated epithelial cells of jejunum in sucking piglet and promoted the polyamine metabolism, however, no statistically significant effect was observed on crypt cell. In addition, exogenous putrescine and proline promote differentiation in the intestinal epithelial cells by Wnt signaling pathways.
