子宫内慢性缺氧对子代大鼠胰腺Pdx-1基因表观遗传机制改变的研究
发布时间：2019-06-30 22:39
【摘要】：目的探讨SD大鼠妊娠期子宫内慢性缺氧(CIH)后,其子代大鼠胰腺Pdx-1基因表达水平。方法选择处于妊娠期的16只健康SD大鼠为研究对象,按照随机数字表法,将其随机分为CIH组及对照组,每组各8只。对CIH组孕鼠建立CIH模型,对照组孕鼠不进行CIH处理。CIH组孕鼠第1天缺氧1h后,对2组孕鼠均进行动脉血气分析。2组SD大鼠各分娩子代大鼠192与208只。于日龄为56d时,采用随机数字表法随机选取CIH组及对照组子代大鼠各24只,然后处死,摘取胰腺,制作胰腺组织标本,分别纳入CIH亚组与对照亚组。对CIH亚组与对照亚组的各6例子代大鼠胰腺标本,采用苏木精-伊红染色法(HE)染色及光镜观察子代大鼠胰岛细胞改变情况,并采用Image Pro Plus 6.0图像分析软件对胰岛β细胞面积进行定量分析。采用免疫组化、Western blotting法分别检测CIH亚组与对照亚组的各6例子代大鼠胰腺Pdx-1蛋白表达水平。采用亚硫酸氢盐测序法,检测CIH亚组与对照亚组的各6例子代大鼠胰腺Pdx-1基因启动子甲基化水平。采用统计学分析方法比较2组孕鼠动脉血气分析结果,以及子代大鼠胰岛β细胞面积,胰腺Pdx-1蛋白相对积分光密度(IOD)值及其表达水平,胰腺Pdx-1基因启动子甲基化水平。结果①2组孕鼠动脉血气分析结果显示,CIH组孕鼠动脉血氧饱和度(SaO_2)、动脉血氧分压(PaO_2)水平,均较对照组显著下降,差异均有统计学意义(t=9.345,P0.001;t=2.537,P=0.028)。2组孕鼠动脉血二氧化碳分压(PaCO_2)及动脉血pH值分别比较,差异均无统计学意义(t=0.228,P=0.822;t=0.100,P=0.920)。②光镜下观察2组子代大鼠胰腺组织切片(HE染色)结果显示,CIH亚组子代大鼠胰岛细胞大小不一,多数胰岛细胞增生肥大;对照亚组子代大鼠胰岛细胞呈圆形或椭圆形的细胞团,胰岛细胞分布均匀。CIH亚组子代大鼠胰岛β细胞面积为(254 216±6 211)μm~2,较对照亚组的(208 253±2 714)μm~2明显增大,二者比较,差异有统计学意义(t=10.313,P0.001)。③免疫组化结果显示,CIH亚组子代大鼠胰腺Pdx-1蛋白的相对IOD值为(1.84±0.51)×10~4,显著低于对照亚组的(3.61±0.71)×10~4,二者比较,差异有统计学意义(t=14.177,P0.001)。Western blotting法检测结果显示,CIH亚组子代大鼠胰腺Pdx-1蛋白表达水平为(1.08±0.22)mg/(kg·d),较对照亚组的(2.09±0.36)mg/(kg·d)低,二者比较,差异有统计学意义(t=0.787,P=0.040)。④2个亚组子代大鼠胰腺CpG岛1、2甲基化水平分别比较,差异均无统计学意义(P0.05)。CIH亚组子代大鼠胰腺CpG岛3甲基化水平显著高于对照亚组,并且差异有统计学意义(t=17.626,P0.001)。结论 SD大鼠产前发生CIH,可导致子代大鼠胰Pdx-1基因启动子甲基化表达上调,Pdx-1基因表达下降,Pdx-1蛋白表达减少。
[Abstract]:Objective to investigate the expression of Pdx-1 gene in pancreas of SD rats after chronic hypoxia (CIH) during pregnancy. Methods Sixteen healthy SD rats during pregnancy were randomly divided into CIH group (n = 8) and control group (n = 8). The CIH model of pregnant rats in CIH group was established, while the pregnant rats in control group were not treated with CIH. The pregnant rats in cih group were exposed to hypoxia for 1 hour on the first day, and the arterial blood gas analysis was performed in both groups. There were 192 and 208 SD rats in each offspring of the two groups. At the age of 56 days, 24 rats in CIH group and 24 offspring in control group were randomly selected by random digital table method, then killed, pancreas was extracted, pancreatic tissue specimens were made and included into CIH subgroup and control subgroup, respectively. The pancreatic specimens of 6 offspring rats in CIH subgroup and control subgroup were observed by (HE) staining and light microscope, and the area of islet 尾 cells was quantitatively analyzed by Image Pro Plus 6.0 image analysis software. The expression of Pdx-1 protein in pancreas of 6 offspring of CIH subgroup and 6 offspring of control subgroup was detected by immunohistochemical, Western blotting method. The methylation level of pancreatic Pdx-1 gene promoter in 6 offspring of CIH subgroup and control subgroup was detected by bisulfite sequencing. The results of arterial blood gas analysis, the area of islet 尾 cells, the relative integral optical density (IOD) of pancreatic Pdx-1 protein and its expression, and the methylated level of pancreatic Pdx-1 gene promoter were compared by statistical analysis. Results the results of arterial blood gas analysis in 12 groups showed that the levels of arterial oxygen saturation (SaO_2) and arterial partial pressure of oxygen (PaO_2) in CIH group were significantly lower than those in control group (t 鈮，
本文编号：2508313
[Abstract]:Objective to investigate the expression of Pdx-1 gene in pancreas of SD rats after chronic hypoxia (CIH) during pregnancy. Methods Sixteen healthy SD rats during pregnancy were randomly divided into CIH group (n = 8) and control group (n = 8). The CIH model of pregnant rats in CIH group was established, while the pregnant rats in control group were not treated with CIH. The pregnant rats in cih group were exposed to hypoxia for 1 hour on the first day, and the arterial blood gas analysis was performed in both groups. There were 192 and 208 SD rats in each offspring of the two groups. At the age of 56 days, 24 rats in CIH group and 24 offspring in control group were randomly selected by random digital table method, then killed, pancreas was extracted, pancreatic tissue specimens were made and included into CIH subgroup and control subgroup, respectively. The pancreatic specimens of 6 offspring rats in CIH subgroup and control subgroup were observed by (HE) staining and light microscope, and the area of islet 尾 cells was quantitatively analyzed by Image Pro Plus 6.0 image analysis software. The expression of Pdx-1 protein in pancreas of 6 offspring of CIH subgroup and 6 offspring of control subgroup was detected by immunohistochemical, Western blotting method. The methylation level of pancreatic Pdx-1 gene promoter in 6 offspring of CIH subgroup and control subgroup was detected by bisulfite sequencing. The results of arterial blood gas analysis, the area of islet 尾 cells, the relative integral optical density (IOD) of pancreatic Pdx-1 protein and its expression, and the methylated level of pancreatic Pdx-1 gene promoter were compared by statistical analysis. Results the results of arterial blood gas analysis in 12 groups showed that the levels of arterial oxygen saturation (SaO_2) and arterial partial pressure of oxygen (PaO_2) in CIH group were significantly lower than those in control group (t 鈮，
本文编号：2508313
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