地塞米松治疗孕鼠肝内胆汁淤积症中的调控机制探讨

发布时间：2018-06-16 06:12

本文选题：地塞米松 + 孕激素　；参考：《新乡医学院》2017年硕士论文

【摘要】：背景妊娠期肝内胆汁淤积症(Intrahepatic Cholestasis of Pregnancy,ICP)是孕期发生的一种临床综合症,如:孕妇肝损伤出现,肝酶升高,总胆酸异常,胎儿、新生儿缺氧死亡发生等。既往研究表明该病可诱使复杂的胆汁酸基因调控网络发生异常,继发围产儿不良预后结局出现,并且通过现有临床技术手段难以提前预测,找到一种较好的治疗方法或安全有效的预防方式是产科有待解决的一大难题。目的探讨法尼醇受体(Farnesoid X receptor,FXR)/胆盐输出泵(Bile salt export pump,BSEP)对孕激素诱导的妊娠期SD大鼠肝内胆汁淤积症的调控作用及地塞米松的干预机制。方法选择清洁级SD孕鼠45只,随机区组分成3组(阴性对照组:对照组;阳性对照组:模型组;治疗组),每组15只,妊娠第10-14天,每天腹腔注射1次:对照组生理盐水2.5ml/kg/d;模型组及治疗组注射孕酮225mg/kg/d建立动物模型。各组孕鼠于妊娠第10天及第15天分别采心脏血2ml,测定血生化指标。治疗组造模成功后于妊娠d15-19天,每天1次大腿内侧肌肉注射地塞米松1mg/kg,妊娠第20天采集心脏血2mL备用。3组孕鼠出现临产征兆时立即行剖宫终止妊娠,记录胎鼠数,死胎数,测量胎鼠体重;取孕鼠肝脏组织观察病理学变化,采用逆转录聚合酶链反应技术检测FXR、BSEP两者mRNA的表达,蛋白质印迹法检测两者蛋白的表达。结果(1)各组孕鼠血生化指标用药前差异无统计学意义(P0.05),模型组用药后血清总胆酸(Total bile acid,TBA)及谷丙转氨酶(Alanine aminotransferase,ALT)明显升高(P0.05),地塞米松干预前后比较,差异有统计学意义(P0.05)。(2)HE染色:模型组孕鼠汇管区肝细胞出现水肿,肝血窦间隙变窄,少许肝细胞出现脂肪变性;治疗组孕鼠肝细胞排列整齐无明显水肿,无脂肪变性,肝血窦间隙正常。(3)死胎率比较:模型组显著高于对照组,治疗组显著低于模型组(P0.05)。(4)各组胎鼠体重比较:模型组明显低于对照组和治疗组,差异有统计学意义(P0.05)。(5)模型组孕鼠肝脏中FXR、FXRmRNA的表达上调,BSEP、BSEPmRNA的表达下调(P0.05);治疗组与模型组比较孕鼠肝脏中FXR、FXRmRNA表达下调,BSEP、BSEPmRNA的表达上调(P0.05)。结论地塞米松干预ICP的调控机制可能是通过减少了妊娠大鼠肝脏FXR的表达,上调BSEP的表达,降低了机体对孕激素及其代谢产物的敏感性,从而对胆汁淤积孕鼠的肝脏功能起到了保护作用,并有效减少胎鼠不良预后的出现。
[Abstract]:Intrahepatic Cholestasis of Pregnancy (ICP) is a clinical syndrome occurring during pregnancy, such as the occurrence of liver injury in pregnant women, the increase of liver enzyme, the abnormal total cholic acid, fetal and neonatal hypoxia death. Previous studies have shown that the disease can induce the complex bile acid gene regulation network to occur abnormality. Poor prognosis of perinatal fetus appears, and it is difficult to predict in advance through existing clinical techniques. Finding a better treatment method or safe and effective prevention is a major problem to be solved in obstetrics. The purpose of this study is to explore the Farnesoid X receptor (FXR) / bile salt output pump (Bile salt export pump, BSEP) for pregnancy The regulation of intrahepatic cholestasis of SD rats and the intervention mechanism of dexamethasone. Methods 45 clean SD pregnant rats were selected and divided into 3 groups (negative control group: control group; positive control group: model group; treatment group), 15 rats in each group, 10-14 days of pregnancy, 1 times per day: saline 2.5ml/kg in control group. /d, the model group and the treatment group were injected with progesterone 225mg/kg/d to establish an animal model. Each group of pregnant rats took the heart blood 2ml for the tenth and 15 days of pregnancy, respectively, and measured the blood biochemical indexes. After the success of the treatment group, 1 times every day, 1 times the medial thigh muscles were injected with dexamethasone 1mg/kg, and the 2mL spare.3 group of the heart blood was collected for twentieth days of pregnancy. The gestational termination of pregnancy was performed immediately, the number of fetal rats, the number of stillbirths, the weight of fetal rats were recorded, the pathological changes of the liver tissues of pregnant rats were observed. The expression of FXR, BSEP mRNA were detected by reverse transcription polymerase chain reaction (RT), and the expression of egg white was detected by Western blot. Results (1) the blood biochemical indexes of pregnant rats were used before drug use. The difference was not statistically significant (P0.05). The serum total cholic acid (Total bile acid, TBA) and alanine transaminase (Alanine aminotransferase, ALT) in the model group were significantly increased (P0.05). The difference was statistically significant (P0.05) before and after the intervention of dexamethasone. (2) HE staining: the liver cells of the model group of pregnant rats were edema and the hepatic sinusoidal space narrowed, In the treatment group, the liver cells in the treatment group had no obvious edema, no fat degeneration, and the hepatic sinusoidal space was normal. (3) the rate of stillbirth was compared: the model group was significantly higher than the control group, and the treatment group was significantly lower than the model group (P0.05). (4) the weight comparison of the mice in each group was significantly lower than the control group and the treatment group, the difference was statistically significant. (5) (5) the expression of FXR, FXRmRNA, BSEP and BSEPmRNA was down regulated in the liver of the model group, and the expression of FXR, FXRmRNA, BSEP and BSEPmRNA in the liver of the pregnant rats was down regulated (P0.05) in the treatment group. Expression, up - regulation the expression of BSEP, reduce the body's sensitivity to progestin and its metabolites, thus protecting the liver function of gestation rats, and effectively reducing the occurrence of bad prognosis in fetal mice.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.255

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