子痫前期对滋养细胞脂肪酸氧化的影响及其与p38MAPK信号通路的相关性  

Interaction mechanism and influence between fatty acid oxidation in trophoblast cells and p38MAPK signal transduction pathway of severe preeclampsia

作　　者：孙晓乐[1] 杨孜[1] 王威[1] 王晓晔[1] 王伽略[1] 武淑英[1] 

机构地区：[1]北京大学第三医院妇产科,100191

出　　处：《中华妇产科杂志》2013年第11期853-857,共5页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金（30973204）;北京市自然科学基金（7102162）

摘　　要：目的 探讨重度子痫前期对胎盘滋养细胞线粒体长链脂肪酸氧化酶——长链三羟基酰基辅酶A脱氢酶（LCHAD）表达的影响及其与p38丝裂原活化蛋白激酶（p38 MAPK）信号通路的相关性.方法 体外培养胎盘滋养细胞,分别用早发型重度子痫前期、晚发型重度子痫前期、重度子痫前期伴发HELLP综合征及正常妊娠妇女的血清刺激胎盘滋养细胞（分别命名为早发组、晚发组、HELLP组、正常对照组）,每组再分别加入DMEM/F12培养基、还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶抑制剂（NADPH-Ⅰ）和p38MAPK抑制剂（p38-Ⅰ）处理细胞.采用实时荧光定量PCR技术和蛋白印迹法检测各组滋养细胞LCHAD mRNA和蛋白的表达.结果 （1）LCHAD mRNA的表达：正常对照组＋培养基、早发组＋培养基、早发组＋ NADPH-Ⅰ、早发组＋p38-Ⅰ、晚发组＋培养基、晚发组＋NADPH-Ⅰ、晚发组＋p38-Ⅰ、HELLP组＋培养基、HELLP组＋NADPH-Ⅰ、HELLP组＋p38-Ⅰ滋养细胞LCHAD mRNA的相对表达量分别为1.00±0.03、0.14±0.08、0.95±0.20、1.43±1.02、0.37±0.18、1.51 ±0.36、1.60±0.31、0.10 ±0.04、0.49 ±0.10、0.44±0.21.早发组＋培养基、晚发组＋培养基、HELLP组＋培养基与正常对照组＋培养基比较,LCHAD mRNA的相对表达量均明显降低,分别比较,差异均有统计学意义（P＜0.05）;与正常对照组比较,晚发组＋ NADPH-Ⅰ、晚发组＋p38-Ⅰ LCHADmRNA的相对表达量均有明显升高,差异均有统计学意义（P＜0.05）,而HELLP组LCHAD mRNA的相对表达量均有明显降低,差异也有统计学意义（P＜0.05）.（2） LCHAD蛋白的表达：正常对照组＋培养基、早发组＋培养基、早发组＋ NADPH-Ⅰ、早发组＋p38-Ⅰ、晚发组＋培养基、晚发组＋NADPH-Ⅰ、晚发组＋ p38-Ⅰ、HELLP组＋培养基、HELLP组＋NADPH-Ⅰ、HELLP组＋p38-Ⅰ组LCHAD蛋白的相对表达量分别为19.4±2.2、10.7±1.1、17.9±3.3、19.1 ±2.9、16.4±2Objective To investigate the effects of expression of mitochondria long-chain fatty acid oxidative enzyme （long-chain 3 hyroxyacyl CoA dehydrogenase,LCHAD） and p38 mitogen activated proteinkinase （p38MAPK） signal transduction pathway in severe preeclampsia.Methods Serum-free trophoblast cells cultured in vitro were stimulated by early onset severe preeclampsia serum （E-PE group）,late onset severe preeclampsia serum （L-PE group）,HELLP syndrome serum （HELLP group）,and normal pregnancy serum （NP group） respectively; each group was added DMEM/F12 medium,reduced form of nicotinamide adenine dinucleotide phosphate （NADPH） oxidase inhibitor （NADPH-Ⅰ） and p38 MAPK inhibitor （p38-Ⅰ）to stimulate cells.Expression of mRNA and protein of LCHAD in trophoblast cells were detected by real-time PCR and western blot.Results （1） The expression of mRNA of LCHAD：the level of mRNA of LCHAD in NP＋DMEM,E-PE ＋ DMEM,E-PE ＋ NADPH-Ⅰ,E-PE ＋ p38-Ⅰ,L-PE ＋ DMEM,L-PE ＋ NADPH-Ⅰ,L-PE ＋ p38-Ⅰ and HELLP ＋ DMEM,HELLP ＋ NADPH-Ⅰ,HELLP ＋ p38-Ⅰ groups were 1.00 ± 0.03,0.14 ±0.08,0.95 ±0.20,1.43±1.02,0.37 ±0.18,1.51 ±0.36,1.60 ±0.31,0.10 ±0.04,0.49 ±0.10,0.44 ± 0.21,respectively.The relative expressions of mRNA of LCHAD were significantly reduced in E-PE ＋ DMEM,L-PE ＋ DMEM and HELLP ＋ DMEM groups compared with the NP ＋ DMEM group （P ＜0.05）.Compared with the NP groups,the relative expressions of mRNA of LCHAD were significantly increased in L-PE ＋ NADPH-Ⅰ and L-PE ＋ p38-Ⅰ group （P ＜ 0.05）,while reduced in HELLP groups （P ＜0.05）.（2） The expression of protein of LCHAD：the relative expressions of protein of LCHAD in NP ＋DMEM,E-PE ＋ DMEM,E-PE ＋ NADPH-Ⅰ,E-PE ＋ p38-Ⅰ,L-PE ＋ DMEM,L-PE ＋ NADPH-Ⅰ,L-PE ＋p38-Ⅰ and HELLP ＋ DMEM,HELLP ＋ NADPH-Ⅰ,HELLP ＋ p38-Ⅰ groups were 19.4 ± 2.2,10.7 ± 1.1,17.9±3.3,19.1 ±2.9,16.4 ±2.3,20.3 ±2.3,20.9 ±4.3,12.4 ±2.3,17.6 ±2.6,17.7 ±2.0 respectively.Compared with the NP

关 键 词：先兆子痫 滋养层 3-羟酰CoA脱氢酶类 NADPH P38丝裂原活化蛋白激酶类 脂肪酸氧化 

分 类 号：R743[医药卫生—神经病学与精神病学]