孕鼠肝内胆汁淤积症对新生鼠肺水通道蛋白3的影响

Effects of Intrahepatic Cholestasis in Pregnant Rats on Pulmonary Aquaporin 3 in Fetal Rats.

目的探讨孕鼠肝内胆汁淤积症对新生鼠肺水通道蛋白3表达的影响。方法健康清洁级孕龄14日的SD大鼠随机分成ICP组和对照组。ICP组利用17α-乙炔基雌二醇(EE)诱导处理建立妊娠期肝内胆汁淤积症大鼠模型。于预产当日行剖宫产娩出新生鼠,并断头处死孕鼠。取新生鼠肺组织进行干湿重测定,HE染色行Holfbauer病理评分,TUNEL检测细胞凋亡,免疫荧光、Western-blotting、荧光定量PCR检查AQP3表达。结果ICP组新生鼠肺脏湿/干比(3.27±0.35)与对照组(1.98±0.07)相比显著增加,差异有统计学意义(P<0.01)。ICP组Holfbauer病理评分(1.60±0.52)分与对照组(0.80±0.63)分相比明显增加,差异有统计学意义(P<0.05)。ICP组凋亡指数(10.31±5.35)%较对照组(1.03±0.47)%明显升高,差异有统计学意义(P<0.01)。免疫荧光染色、Western-blotting、荧光定量PCR检测均提示ICP组新生鼠肺AQP3表达较对照组明显下降(P<0.01)。以新生鼠肺细胞凋亡率为一变量,肺组织免疫荧光检测AQP3表达量(IOD值)为另一变量进行相关性分析,结果显示新生鼠肺组织凋亡与肺细胞AQP3表达量呈负相关(R=-0.621,P<0.01)。结论孕鼠肝内胆汁淤积症可导致新生鼠肺水通道蛋白3的表达下降,细胞凋亡增加。

Objective To investigate the effect of intrahepatic cholestasis in pregnant rats on the expression of aquaporin 3 in neonatal rat lung.Method Healthy SD rats at 14 days of gestational age were randomly divided into ICP group and control group.The ICP group was treated with 17α-acetylenestradiol(EE)to establish the model of intrahepatic cholestasis during pregnancy.The newborn rats were delivered by cesarean section on the day of pre-delivery,and the pregnant rats were killed by decapitation.Lung tissue of neonatal rats was collected for dry and wet weight measurement,and Holfbauer pathological score was obtained by HE staining.Cell apoptosis was detected by TUNEL,and the expression of AQP3 was detected by immunofluorescence,Western-blotting and fluorescence quantitative PCR.Results Compared with the control group(1.98±0.07),the lung wet/dry ratio of neonatal rats in ICP group was significantly increased(3.27±0.35),and the difference was statistically significant(P<0.01).Holfbauer pathological score in ICP group(1.60±0.52)was significantly higher than that in control group(0.80±0.63),and the difference was statistically significant(P<0.05).The apoptosis index of ICP group(10.31±5.35)%was significantly higher than that of control group(1.03±0.47)%,and the difference was statistically significant(P<0.01).Immunofluorescence staining,Western Blot and quantitative PCRshowed that the expression of AQP3 in neonatal lung of ICP group was significantly lower than that of control group(P<0.01).Using the apoptosis rate of neonatal lung cells as one variant and the expression of AQP3(IOD value)in lung tissue as the other variant,the correlation analysis showed that the apoptosis of neonatal lung tissue was negatively correlated with the expression of AQP3 in lung cells(r=-0.621,P<0.01).Conclusion Intrahepatic cholestasis in pregnant rats can lead to decreased expression of aquaporin 3 in neonatal rat lung,with increased apoptosis.