摘要
目的:(1)分析神经肽 Y(NPY)和血红素加氧酶-1(HO-1)在妊娠期肝内胆汁淤积症(ICP)孕鼠胎盘中表达及 NPY 在胎鼠脑组织的表达,探讨 ICP 的发病机制。(2)分析 NPY 和 HO-1 在 ICP 孕妇胎盘中的表达和血清甘胆酸的关系,进一步探讨 NPY、HO-1 在 ICP 中作用。
方法:(1)建立孕鼠 ICP 模型,观察孕鼠肝脏和胎盘的病理变化;应用免疫组织化学染色方法及图像分析技术检测 ICP 孕鼠(10 例,实验组)及正常孕鼠(10 例,对照组)胎盘中 NPY、HO-1 表达及定量;NPY 在胎鼠脑组织中的表达及定量(2)收集 ICP 孕妇胎盘标本,检测轻度 ICP(20 例)、重度 ICP(20 例)、正常妊娠组(30 例)胎盘中 NPY和 HO-1 的表达。(3)用放射免疫法测定三组孕妇血清甘胆酸水平。
结果:①孕鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆汁酸(TBA)明显升高;肝细胞水肿,胞质内出现空泡,毛细胆管扩张、胆汁淤积;胎盘绒毛肿胀、增粗,绒毛间隙变窄,滋养细胞增生。②ICP 孕鼠组胎盘 NPY 免疫反应呈强阳性;对照组部分滋养细胞的 NPY 免疫反应阳性,绒毛的毛细血管内皮细胞呈弱阳性或阴性。图像分析结果显示:ICP 组 NPY 平均光密度值(OD 值)为(1.6421±0.0412)高于对照组(0.7820±0.0229),两组比较有统计学意义 P<0.01。ICP 孕鼠组胎盘部分滋养细胞的 HO-1 呈弱阳性或阴性,对照组 HO-1免疫反应呈阳至强阳性。图像分析结果显示:ICP 组 HO-1 的 OD 值
Objective:(1) to detect the expression of neuropeptideY (NPY) and heme oxygenase-1 (HO-1) in placentas of rats with intrahepatic cholestasis of pregnancy (ICP) and the expression of NPY in fetal brains of rats with ICP, to explore the reasons of intrauterine growth retardation and intrauterine fetal death in ICP. (2) to analyze the correlation between serum levels of glycocholic acid (CG) in patients with ICP and expression level of NPY、HO-1 in placentas of patients with ICP.
Materials and Methods :(1): Rats fifteen days pregnant were administered Estrogen and Progesterone to induce ICP. The structural changes of liver and placenta of the pregnant rats were observed. And biochemical tests of blood were conducted. The placentas of normal pregnant rats (n=10) and ICP (n=10) were used for detecting expression and quantification of NPY and HO-1 by immunohistochemistry staining and image analysis. The fetal brains of normal pregnant rats (n=10) and ICP (n=10) were used for detecting expression and quantification of NPY
by immunohistochemistry staining and image analysis.(2) the placentas of patients with ICP were collected . The placentas of normal pregnancy (n=30)、mild ICP(n=20) and severe ICP(n=20) were used for detecting expression and quantification of NPY and HO-1 by immunohistochemistry staining and image analysis.(3) Serum levels of CG in these three groups were examined by radio-immunoassay before delivery.
result:(1) in the experimental group ,the levels of serum total bile salt (TBA), alanine aminotransferase (ALT), aspartate transaminase(AST) were significantly increased P〈0.05, structural changes of hepatocytes of ICP rats include swelling , bubbling ,capillary expanding ,dilation of the bile canaliculi. The placentas of ICP showed the villis were swelling , the size of intervilli space was smaller ,the trophocytes were hyperplastic.(2)Almost all placenta villis of ICP group showed NPY strong positive immunoreaction ,but in control group ,some trophocytes appeared NPY positive ,and capillary endothelium in placental villi showed NPY weak positive or negative. average value of optical density(OD) of NPY by image analysis in ICP group and in the control group was (1.6421±0.0412),(0.7820±0.0229),respectively. The OD value in ICP group was significantly higher than that in the control group P<0.01. Placenta villis of ICP group showed HO-1 weak positive or negative immunoreaction, but in control group, some trophocytes appeared positive. average value of optical density(OD) of HO-1 by image analysis in ICP group and in the
control group was (0.9264±0.0250), (1.3271±0.0178),respectively. The OD value in ICP group was significantly lower than that in the control group P<0.05. (3) Fetal brain tissues of ICP group showed NPY positive immunoreaction, but showed negative in control group. average value of optical density(OD) of NPY in ICP group and in the control group was (1.0591±0.0185), (0.6302±0.0259),respectively. The OD value in ICP group was significantly higher than that in the control group P<0.05. (4) Placenta villis of patients with ICP showed NPY immunoreaction was increased with the staging of disease . average value of optical density(OD) of NPY in normal pregnancy, mild ICP and severe ICP was (0.7230±0.1452)、(1.0873±0.1394)、(1.6300±0.1281),respectively (P>0.05 between normal pregnancy and mild ICP, P<0.01 between normal and severe ICP, P<0.05 between mild and severe; HO-1 immunoreaction of placenta villis was lightly increased in mild ICP than normal, but was significantly reduced in severe ICP. average value of optical density(OD) of HO-1 in normal pregnancy、mild ICP and severe ICP was(1.1025±0.1106)、(1.3532±0.1208)、(0.7393±0.1008),respectively P>0.05 between normal pregnancy and mild ICP, P<0.01between normal and severe ICP, P<0.05between mild and severe . (5) Serum level of CG in normal pregnancy、mild ICP and severe ICP was 142.5±80.1 ug/dl,1498.5±948.57 ug/dl,1869.6±954.02 ug/dl, respectively. A significant positive correlation between CG level and NPY expression level (r=0.4
引文
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