中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2013年
11期
853-857
,共5页
孙晓乐%杨孜%王威%王晓晔%王伽略%武淑英
孫曉樂%楊孜%王威%王曉曄%王伽略%武淑英
손효악%양자%왕위%왕효엽%왕가략%무숙영
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
先兆子癇%滋養層%3-羥酰CoA脫氫酶類%NADPH%p38絲裂原活化蛋白激酶類%脂肪痠氧化
선조자간%자양층%3-간선CoA탈경매류%NADPH%p38사렬원활화단백격매류%지방산양화
Pre-eclampsia%Trophoblasts%3-hyroxyacyl CoA dehydrogenases%NADPH%p38 mitogen-activated protein kinases%Fatty acid oxidation
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±2.3、20.3±2.3、20.9 ±4.3、12.4±2.3、17.6±2.6、17.7 ±2.0.与正常对照组比较,早发组+培养基、晚发组+培养基及HELLP组LCHAD蛋白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),而早发组+NADPH-Ⅰ、早发组+p38-Ⅰ、晚发组+NADPH-Ⅰ、晚发组+p38-Ⅰ LCHAD蛋白的相对表达量无明显变化(P＞0.05).早发组、晚发组、HELLP组中,NADPH-Ⅰ亚组、p38-Ⅰ亚组与培养基亚组比较,LCHAD蛋白的相对表达量均有明显升高,差异均有统计学意义(P＜0.05).结论 重度子痫前期对胎盘滋养细胞中的脂肪酸氧化代谢有一定的影响,尤其以HELLP综合征最为明显.NADPH-Ⅰ、p38-Ⅰ能缓解早、晚发型重度子痫前期和HELLP综合征中的脂肪酸氧化代谢障碍,p38MAPK信号通路可能参与了该过程.
目的 探討重度子癇前期對胎盤滋養細胞線粒體長鏈脂肪痠氧化酶——長鏈三羥基酰基輔酶A脫氫酶(LCHAD)錶達的影響及其與p38絲裂原活化蛋白激酶(p38 MAPK)信號通路的相關性.方法 體外培養胎盤滋養細胞,分彆用早髮型重度子癇前期、晚髮型重度子癇前期、重度子癇前期伴髮HELLP綜閤徵及正常妊娠婦女的血清刺激胎盤滋養細胞(分彆命名為早髮組、晚髮組、HELLP組、正常對照組),每組再分彆加入DMEM/F12培養基、還原型煙酰胺腺嘌呤二覈苷痠燐痠氧化酶抑製劑(NADPH-Ⅰ)和p38MAPK抑製劑(p38-Ⅰ)處理細胞.採用實時熒光定量PCR技術和蛋白印跡法檢測各組滋養細胞LCHAD mRNA和蛋白的錶達.結果 (1)LCHAD mRNA的錶達:正常對照組+培養基、早髮組+培養基、早髮組+ NADPH-Ⅰ、早髮組+p38-Ⅰ、晚髮組+培養基、晚髮組+NADPH-Ⅰ、晚髮組+p38-Ⅰ、HELLP組+培養基、HELLP組+NADPH-Ⅰ、HELLP組+p38-Ⅰ滋養細胞LCHAD mRNA的相對錶達量分彆為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.早髮組+培養基、晚髮組+培養基、HELLP組+培養基與正常對照組+培養基比較,LCHAD mRNA的相對錶達量均明顯降低,分彆比較,差異均有統計學意義(P＜0.05);與正常對照組比較,晚髮組+ NADPH-Ⅰ、晚髮組+p38-Ⅰ LCHADmRNA的相對錶達量均有明顯升高,差異均有統計學意義(P＜0.05),而HELLP組LCHAD mRNA的相對錶達量均有明顯降低,差異也有統計學意義(P＜0.05).(2) LCHAD蛋白的錶達:正常對照組+培養基、早髮組+培養基、早髮組+ NADPH-Ⅰ、早髮組+p38-Ⅰ、晚髮組+培養基、晚髮組+NADPH-Ⅰ、晚髮組+ p38-Ⅰ、HELLP組+培養基、HELLP組+NADPH-Ⅰ、HELLP組+p38-Ⅰ組LCHAD蛋白的相對錶達量分彆為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.與正常對照組比較,早髮組+培養基、晚髮組+培養基及HELLP組LCHAD蛋白的相對錶達量均明顯降低,分彆比較,差異均有統計學意義(P＜0.05),而早髮組+NADPH-Ⅰ、早髮組+p38-Ⅰ、晚髮組+NADPH-Ⅰ、晚髮組+p38-Ⅰ LCHAD蛋白的相對錶達量無明顯變化(P＞0.05).早髮組、晚髮組、HELLP組中,NADPH-Ⅰ亞組、p38-Ⅰ亞組與培養基亞組比較,LCHAD蛋白的相對錶達量均有明顯升高,差異均有統計學意義(P＜0.05).結論 重度子癇前期對胎盤滋養細胞中的脂肪痠氧化代謝有一定的影響,尤其以HELLP綜閤徵最為明顯.NADPH-Ⅰ、p38-Ⅰ能緩解早、晚髮型重度子癇前期和HELLP綜閤徵中的脂肪痠氧化代謝障礙,p38MAPK信號通路可能參與瞭該過程.
목적 탐토중도자간전기대태반자양세포선립체장련지방산양화매——장련삼간기선기보매A탈경매(LCHAD)표체적영향급기여p38사렬원활화단백격매(p38 MAPK)신호통로적상관성.방법 체외배양태반자양세포,분별용조발형중도자간전기、만발형중도자간전기、중도자간전기반발HELLP종합정급정상임신부녀적혈청자격태반자양세포(분별명명위조발조、만발조、HELLP조、정상대조조),매조재분별가입DMEM/F12배양기、환원형연선알선표령이핵감산린산양화매억제제(NADPH-Ⅰ)화p38MAPK억제제(p38-Ⅰ)처리세포.채용실시형광정량PCR기술화단백인적법검측각조자양세포LCHAD mRNA화단백적표체.결과 (1)LCHAD mRNA적표체:정상대조조+배양기、조발조+배양기、조발조+ NADPH-Ⅰ、조발조+p38-Ⅰ、만발조+배양기、만발조+NADPH-Ⅰ、만발조+p38-Ⅰ、HELLP조+배양기、HELLP조+NADPH-Ⅰ、HELLP조+p38-Ⅰ자양세포LCHAD mRNA적상대표체량분별위1.00±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.조발조+배양기、만발조+배양기、HELLP조+배양기여정상대조조+배양기비교,LCHAD mRNA적상대표체량균명현강저,분별비교,차이균유통계학의의(P＜0.05);여정상대조조비교,만발조+ NADPH-Ⅰ、만발조+p38-Ⅰ LCHADmRNA적상대표체량균유명현승고,차이균유통계학의의(P＜0.05),이HELLP조LCHAD mRNA적상대표체량균유명현강저,차이야유통계학의의(P＜0.05).(2) LCHAD단백적표체:정상대조조+배양기、조발조+배양기、조발조+ NADPH-Ⅰ、조발조+p38-Ⅰ、만발조+배양기、만발조+NADPH-Ⅰ、만발조+ p38-Ⅰ、HELLP조+배양기、HELLP조+NADPH-Ⅰ、HELLP조+p38-Ⅰ조LCHAD단백적상대표체량분별위19.4±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.여정상대조조비교,조발조+배양기、만발조+배양기급HELLP조LCHAD단백적상대표체량균명현강저,분별비교,차이균유통계학의의(P＜0.05),이조발조+NADPH-Ⅰ、조발조+p38-Ⅰ、만발조+NADPH-Ⅰ、만발조+p38-Ⅰ LCHAD단백적상대표체량무명현변화(P＞0.05).조발조、만발조、HELLP조중,NADPH-Ⅰ아조、p38-Ⅰ아조여배양기아조비교,LCHAD단백적상대표체량균유명현승고,차이균유통계학의의(P＜0.05).결론 중도자간전기대태반자양세포중적지방산양화대사유일정적영향,우기이HELLP종합정최위명현.NADPH-Ⅰ、p38-Ⅰ능완해조、만발형중도자간전기화HELLP종합정중적지방산양화대사장애,p38MAPK신호통로가능삼여료해과정.
Objective 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 groups,the protein expressions of LCHAD were significantly remarkably reduced in E-PE + DMEM,L-PE + DMEM and HELLP groups (P ＜ 0.05).Compared with the DMEM groups,the protein expressions of LCHAD were significantly increased in NADPH-Ⅰ and p38-Ⅰ groups of E-PE,L-PE and HELLP groups (P ＜ 0.05).Conclusions These studies demonstrate that long chain fatty acid oxidation was involved in the pathogenesis and development of preeclampsia.The expressions of gene and protein of LCHAD were remarkably affected by early onset severe preeclampsia and HELLP syndrome.NADPH-Ⅰ and p38-Ⅰ may allay the disorder of fatty acid oxidation.p38MAPK signal transduction pathway may contributed in this process.
