【作者】 张林；

【导师】 胡雅毅； 单丹；

【作者基本信息】 四川大学 ， 妇产科学（专业学位）， 2023， 硕士

【摘要】 目的:比较妊娠期肝内胆汁淤积症(Intrahepatic Cholestasis of Pregnancy,ICP)患者和正常孕妇胎盘及血浆硒蛋白P(Selenoprotein P,SELP)、硒蛋白S(Selenoprotein S,SELS)的表达差异,探讨SELP、SELS在ICP的表达及临床意义。材料和方法:1.临床样本资料搜集:选择2021年10月至2022年6月在我院产科门诊建卡的单胎择期剖宫分娩孕妇为研究对象,分为对照组(n=22例)、ICP组(n=12例),收集患者的一般资料及临床资料,采集实验对象的胎盘组织。华西附二院标本库提取24例(对照12例,ICP组12例)早、中、晚孕的血浆用于酶联免疫吸附试验(Enzyme-linked Immunosorbent Assay,ELISA)检测。2.实时荧光核酸扩增检测系统(Real-time quantitative PCR detecting system,RT-qPCR)测定两组胎盘组织SELP mRNA、SELS mRNA表达水平。3.蛋白印迹法(Western blotting,WB)测定两组胎盘组织中SELP、SELS的表达量。4.组织免疫组化法(Immunohistochemistry,IHC)确定胎盘组织中SELP、SELS的表达定位。5.酶联免疫吸附试验(ELISA)测量正常对照组及ICP组妊娠早期、中期、晚期血浆SELP、SELS含量变化。结果:1.研究对象临床特征分析结果:ICP组晚孕期ALT、AST、DBIL、GGT、TBA等生化指标均不同程度高于正常对照组。ICP组ALT(P<0.001)、AST(P<0.001)、DBIL(P=0.01)、GGT(P=0.002)、TBA(P<0.001)等生化检测指标的差异具有统计学意义。2.胎盘SELP mRNA在ICP组(0.58±0.39)相比对照组(0.87±0.68)降低,但两者之间差异无统计学差异(P>0.05);胎盘SELS mRNA在ICP组(2.48±0.77)相比对照组(1.21±1.06)明显升高,差异有统计学意义(P<0.05)。3.ICP组胎盘SELP相比对照组表达减少,差异具有统计学意义(P<0.05)。ICP组胎盘SELS相比对照组表达减少,差异具有统计学意义(P<0.01)。4.胎盘SELP主要位于胎盘合体滋养细胞胞质,表达量较少;SELS多位于胎盘合体滋养细胞胞质及包膜中,ICP组表达少于对照组。5.早孕期ICP组血浆SELP含量为(7.9±3.3)mg/L,对照组血浆SELP含量为(8.1±8.0)mg/L,两组间差异比较无统计学差异(P>0.05);中孕期ICP组血浆SELP含量为(5.7±2.7)mg/L,对照组血浆SELP含量为(4.3±1.9)mg/L,两组间差异比较无统计学差异(P>0.05);晚孕期ICP组血浆SELP含量为(7.8±3.7)mg/L,对照组血浆SELP含量为(8.0±3.5)mg/L,两组间差异比较无统计学差异(P>0.05)。整个孕期血浆SELP含量呈中孕期降低,晚孕期升高趋势。6.早孕期ICP组血浆SELS含量为(43.7±2.7)pg/ml,对照组血浆SELS含量为(42.5±1.6)pg/ml,两组间差异比较无统计学差异(P>0.05);中孕期ICP组血浆SELS含量为(52.4±16.5)pg/ml,对照组血浆SELS含量为(42.7±3.4)pg/ml,ICP组高于对照组,两组间差异比较有统计学差异(P<0.05);晚孕期ICP组血浆SELS含量为(46.9±11.7)pg/ml,对照组血浆SELS含量为(42.0±5.1)pg/ml,两组间差异比较无统计学差异(P>0.05)。结论:1.ICP患者晚孕期胎盘SELS表达降低,推测SELS变化与ICP有关联。2.ICP患者晚孕期胎盘SELS mRNA基因转录水平上升,SELS蛋白水平下降。可能与ICP孕妇胎盘循环血氧储备不足,内质网功能受损,导致基因转录水平和蛋白翻译水平不一致,SELS出现合成障碍有关。3.ICP患者妊娠不同时期的血浆SELP、SELS含量与正常对照组相比均无明显变化,与胎盘中SELP、SELS变化并不完全一致,SELP、SELS可能存在组织特异性表达或合成障碍。孕期血浆SELP、SELS含量与ICP的关联还需增加样本量进一步研究。更多还原

【Abstract】 Objective:To compare the difference between the Selenoprotein P(SELP),Selenoprotein S(SELS)in the placenta and plasma of patients with ICP,and to explore the expression and clinical significance of SELP and SELS in ICP.Materials and Methods:1.Collection of the clinical sample:To select the women with single pregnancy who have prenatal examination in the obstetric clinic of our hospital from October2021 to June 2022,divide them into the control group(n=22)and the ICP group(n=12),collect general and clinical data of the patients,and the placental tissue of the experimental subjects.We extracted the plasma of 24 subjects(12 cases in the control group and 12 cases in the ICP group)in our hospital sample bank with different pregnancy periods for Enzyme-linked immunosorbent assay detection.2.Real-time quantitative PCR detection system(RT-qPCR)was used to detect the gene expression level in the placenta of SELP mRNA and SELS mRNA.3.Western blotting(WB)was uesd to determine the expression of the SELP and SELS in placenta.4.Immunohistochemistry(IHC)was used to reveal the localization of the SELP and SELS in placenta.5.ELISA was used to measure the plasma SELP and SELS content in the early,middle and late pregnancy.Results:1.The ALT,AST,DBIL,GGT TBA and other clinical biochemic indexes in late pregnancy ICP group are higher than those in normal control groups.Above all,there were differences in biochemical tests and treatment of ALT(P<0.001),AST(P<0.001),DBIL(P=0.01),GGT(P=0.002),TBA(P<0.001)in ICP group.2.The placental SELP mRNA in the ICP group(0.58±0.39)was lower than that in the control group(0.87±0.68),but with no significant difference(P>0.05).And the placental SELS mRNA in the ICP group(2.48±0.77)was higher than that in the control group(1.21±1.06),with a statistically significant difference between the two groups(P<0.05).3.The expression of SELP in the placenta of ICP group decreased significantly compared with that of normal control group(P<0.05).And the expression of SELS in placenta of ICP was significantly lower than control group(P<0.01).4.The placental SELP is mainly located in the placental syncytiotrophoblast cytoplasm,with a low expression level,while the SELS was mostly located in the placental syncytiotrophoblast cell membrane and cytoplasm.5.Compare to the plasma SELP content duiring early pregnancy in control group(8.1±8.0)mg/L,the plasma SELP content ICP group was(7.9±3.3)mg/L,which was no statistical difference(P>0.05).Compare to the plasma SELP content duiring middle pregnancy in control group(4.3±1.9)mg/L,the plasma SELP content ICP group was(5.7±2.7)mg/L,which was no statistical difference(P>0.05).Compare to the plasma SELP content late middle pregnancy in control group(8.0±3.5)mg/L,the plasma SELP content ICP group was(7.8±3.7)mg/L,which was no statistical difference(P>0.05).During the whole pregnancy,the plasma SELP content decreased in the middle pregnancy but increased in the late pregnancy.6.The plasma SELP content in the early pregnancy ICP group was(43.7±2.7)pg/ml,and the plasma SELS content in control group was(42.5±1.56)pg/ml,with no statistical difference(P>0.05).The plasma SELP content in the ICP group during the middle pregnancy was(52.4±16.5)pg/ml,and the plasma SELP content in the control group was(42.7±3.4)pg/ml.So the ICP group was higher than the control group,with a statistical significance(P<0.05).The plasma SELP content in the late pregnancy ICP group was(46.9±11.7)pg/ml,and that in control group was(42.0±5.1)pg/ml,with no statistical difference(P>0.05).Conclusions:1.The expression of SELS in the placenta in late pregnancy of ICP patients is decreased,suggesting that the changes in SELS may be related to intrahepatic cholestasis during pregnancy.2.The level of SELS mRNA in the placenta in late pregnancy of ICP patients increases,while the level of SELS protein decreases.Due to the insufficient circulating oxygen reserves in the placenta of the ICP pregnant women,the function of the endoplasmic reticulum is impaired,resulting in inconsistent levels of gene transcription protein translation and the SELS synthesis disorders.3.The plasma levels of SELP and SELS in ICP patients have no significant changes compared to the normal control group at different stages of pregnancy,which are inconsistent with the changes in SELP and SELS in the placenta,indicating that there may be tissue specific expression or synthesis disorders of SELP and SELS.The association between the plasma SELP、SELS and the ICP during pregnancy needs to further elaborate through the enlagred sample.更多还原