發(fā)布時(shí)間：2019-06-30 22:39

【摘要】：目的探討SD大鼠妊娠期子宮內(nèi)慢性缺氧(CIH)后,其子代大鼠胰腺Pdx-1基因表達(dá)水平。方法選擇處于妊娠期的16只健康SD大鼠為研究對(duì)象,按照隨機(jī)數(shù)字表法,將其隨機(jī)分為CIH組及對(duì)照組,每組各8只。對(duì)CIH組孕鼠建立CIH模型,對(duì)照組孕鼠不進(jìn)行CIH處理。CIH組孕鼠第1天缺氧1h后,對(duì)2組孕鼠均進(jìn)行動(dòng)脈血?dú)夥治觥?組SD大鼠各分娩子代大鼠192與208只。于日齡為56d時(shí),采用隨機(jī)數(shù)字表法隨機(jī)選取CIH組及對(duì)照組子代大鼠各24只,然后處死,摘取胰腺,制作胰腺組織標(biāo)本,分別納入CIH亞組與對(duì)照亞組。對(duì)CIH亞組與對(duì)照亞組的各6例子代大鼠胰腺標(biāo)本,采用蘇木精-伊紅染色法(HE)染色及光鏡觀察子代大鼠胰島細(xì)胞改變情況,并采用Image Pro Plus 6.0圖像分析軟件對(duì)胰島β細(xì)胞面積進(jìn)行定量分析。采用免疫組化、Western blotting法分別檢測(cè)CIH亞組與對(duì)照亞組的各6例子代大鼠胰腺Pdx-1蛋白表達(dá)水平。采用亞硫酸氫鹽測(cè)序法,檢測(cè)CIH亞組與對(duì)照亞組的各6例子代大鼠胰腺Pdx-1基因啟動(dòng)子甲基化水平。采用統(tǒng)計(jì)學(xué)分析方法比較2組孕鼠動(dòng)脈血?dú)夥治鼋Y(jié)果,以及子代大鼠胰島β細(xì)胞面積,胰腺Pdx-1蛋白相對(duì)積分光密度(IOD)值及其表達(dá)水平,胰腺Pdx-1基因啟動(dòng)子甲基化水平。結(jié)果①2組孕鼠動(dòng)脈血?dú)夥治鼋Y(jié)果顯示,CIH組孕鼠動(dòng)脈血氧飽和度(SaO_2)、動(dòng)脈血氧分壓(PaO_2)水平,均較對(duì)照組顯著下降,差異均有統(tǒng)計(jì)學(xué)意義(t=9.345,P0.001;t=2.537,P=0.028)。2組孕鼠動(dòng)脈血二氧化碳分壓(PaCO_2)及動(dòng)脈血pH值分別比較,差異均無(wú)統(tǒng)計(jì)學(xué)意義(t=0.228,P=0.822;t=0.100,P=0.920)。②光鏡下觀察2組子代大鼠胰腺組織切片(HE染色)結(jié)果顯示,CIH亞組子代大鼠胰島細(xì)胞大小不一,多數(shù)胰島細(xì)胞增生肥大;對(duì)照亞組子代大鼠胰島細(xì)胞呈圓形或橢圓形的細(xì)胞團(tuán),胰島細(xì)胞分布均勻。CIH亞組子代大鼠胰島β細(xì)胞面積為(254 216±6 211)μm~2,較對(duì)照亞組的(208 253±2 714)μm~2明顯增大,二者比較,差異有統(tǒng)計(jì)學(xué)意義(t=10.313,P0.001)。③免疫組化結(jié)果顯示,CIH亞組子代大鼠胰腺Pdx-1蛋白的相對(duì)IOD值為(1.84±0.51)×10~4,顯著低于對(duì)照亞組的(3.61±0.71)×10~4,二者比較,差異有統(tǒng)計(jì)學(xué)意義(t=14.177,P0.001)。Western blotting法檢測(cè)結(jié)果顯示,CIH亞組子代大鼠胰腺Pdx-1蛋白表達(dá)水平為(1.08±0.22)mg/(kg·d),較對(duì)照亞組的(2.09±0.36)mg/(kg·d)低,二者比較,差異有統(tǒng)計(jì)學(xué)意義(t=0.787,P=0.040)。④2個(gè)亞組子代大鼠胰腺CpG島1、2甲基化水平分別比較,差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。CIH亞組子代大鼠胰腺CpG島3甲基化水平顯著高于對(duì)照亞組,并且差異有統(tǒng)計(jì)學(xué)意義(t=17.626,P0.001)。結(jié)論 SD大鼠產(chǎn)前發(fā)生CIH,可導(dǎo)致子代大鼠胰Pdx-1基因啟動(dòng)子甲基化表達(dá)上調(diào),Pdx-1基因表達(dá)下降,Pdx-1蛋白表達(dá)減少。
[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，

本文編號(hào)：2508313