CAJ | 학술논문

目的探讨不同链长游离脂肪酸对人滋养细胞线粒体脂肪酸β氧化循环中长链三羟基酰基辅酶A脱氢酶(LCHAD)基因和蛋白表达变化的影响.方法体外无血清培养人绒毛滋养细胞,将细胞分为五组,每组用不同链长脂肪酸孵育细胞,分别为无游离脂肪酸( F-FFA)、短链脂肪酸(SC-FFA)、中链脂肪酸(MC-FFA)、长链脂肪酸(LC-FFA)、极长链脂肪酸(VLC-FFA).采用实时荧光定量PCR和蛋白印迹法检测各组LCHAD基因和蛋白的表达变化.结果与F-FFA、SC -FFA、MC-FFA 3组比较,LC-FFA组LCHAD基因和蛋白明显降低(P＜0.05),VLC-FFA组LCHAD基因明显升高(P＜0.05),而蛋白表达差异无统计学意义(P＞0.05).F-FFA、SC-FFA、MC-FFA 3组之间LCHAD基因和蛋白表达差异无统计学意义(P＞0.05).与LC-FFA组比较,VLC-FFA组LCHAD基因和蛋白明显升高(P＜0.05).结论游离脂肪酸对人滋养细胞线粒体脂肪酸β氧化循环LCHAD的基因蛋白表达变化存在影响;LC-FFA刺激下的胎盘滋养细胞中存在LC-FFA线粒体β-氧化代谢障碍.VLC-FFA与脂肪酸β-氧化代谢存在一定的相关性,其相互作用机制还有待进一步研究.短链、中链脂肪酸未显示对线粒体脂肪酸β-氧化代谢的影响.
목적 탐토불동련장유리지방산대인자양세포선립체지방산β양화순배중장련삼간기선기보매A탈경매(LCHAD)기인화단백표체변화적영향.방법 체외무혈청배양인융모자양세포,장세포분위오조,매조용불동련장지방산부육세포,분별위무유리지방산( F-FFA)、단련지방산(SC-FFA)、중련지방산(MC-FFA)、장련지방산(LC-FFA)、겁장련지방산(VLC-FFA).채용실시형광정량PCR화단백인적법검측각조LCHAD기인화단백적표체변화.결과 여F-FFA、SC -FFA、MC-FFA 3조비교,LC-FFA조LCHAD기인화단백명현강저(P＜0.05),VLC-FFA조LCHAD기인명현승고(P＜0.05),이단백표체차이무통계학의의(P＞0.05).F-FFA、SC-FFA、MC-FFA 3조지간LCHAD기인화단백표체차이무통계학의의(P＞0.05).여LC-FFA조비교,VLC-FFA조LCHAD기인화단백명현승고(P＜0.05).결론 유리지방산대인자양세포선립체지방산β양화순배LCHAD적기인단백표체변화존재영향;LC-FFA자격하적태반자양세포중존재LC-FFA선립체β-양화대사장애.VLC-FFA여지방산β-양화대사존재일정적상관성,기상호작용궤제환유대진일보연구.단련、중련지방산미현시대선립체지방산β-양화대사적영향.
Objective To explore the interacting mechanisms and influences of different chain lengths of fatty acids and the expression of mitochondria long-chain 3 hydroxyacyl CoA dehydrogenase (LCHAD) in trophoblast cells.Methods The serum-free trophoblast cells cultured in vitro were divided into 5 groups to receieve the stimulations of DMEM/F12 medium without FFA (F-FFA),short-chain fatty acids (SC-FFA),medium-chain fatty acids (MC-FFA),long-chain fatty acids (LC-FFA),very long-chain fatty acids (VLC-FFA).The expressions of mRNA and protein of LCHAD in trophoblast cells were detected by real-time polymerase chain reaction (PCR) and Western blot.Results Compared with the F-FFA,SCFFA and MC-FFA groups,the expressions of gene and protein of LCHAD significantly decreased (P ＜0.05) in the LC-FFA group.The expression of gene of LCHAD increased significantly in the VLC-FFA group (P ＜ 0.05).But no difference existed in protein expression between the VLC-FFA group and other three groups (P ＞ 0.05 ).Gene expression of LCHAD had no difference among the F-FFA,SC-FFA,MCFFA groups ( P ＞ 0.05 ).Compared with the LC-FFA group,the expression of gene of LCHAD increased significantly in the VLC-FFA group ( P ＜ 0.05).Conclusion Free fatty acids may affect the expression of mitochondrial β-oxidation enzyme of LCHAD in trophoblast cells.Long-chain fatty acid alters the LCHAD gene protein expression.The correlation between very long chain fatty acids and the gene expression of LCHAD has been detected and their interactions needs further explorations.Short or medium chain fatty acids have no significant effect on the mitochondrial metabolism of fatty acid β-oxidation in trophoblast cells.