本文選題：鉛　切入點：胎盤　出處：《青島大學》2012年碩士論文　論文類型：學位論文

【摘要】：目的觀察胎盤細胞凋亡、Smad4及S100B蛋白在孕期不同階段鉛暴露下大鼠胎盤中的表達特征及其意義。 方法108只大鼠隨機分為4組,于孕期不同階段飲服0.025%醋酸鉛溶液。原子吸收光譜法測定孕末期血鉛水平。Hoechst細胞核染色以及脫氧核糖核苷酸末端轉(zhuǎn)移酶介導的缺口末端標記(TUNEL)技術細胞凋亡檢測法測定各組不同孕期鉛暴露大鼠胎盤細胞凋亡指數(shù)。免疫組化法測定大鼠各組胎盤中Smad4以及S100B蛋白的表達。計量資料組間比較采用單因素方差分析,計數(shù)資料組間比較采用卡方檢驗。 結果①實驗組血鉛水平均高于0.483μmol/L,不同孕期鉛暴露大鼠實驗組血鉛水平與對照組比較差異有統(tǒng)計學意義(F=12.10,P0.01)。②Hoechst染色顯示,鉛暴露組胎盤滋養(yǎng)層凋亡細胞核濃染,部分核內(nèi)可見致密塊狀熒光顆粒,對照組凋亡細胞呈散在分布。TUNEL技術檢測顯示,鉛暴露組大鼠胎盤滋養(yǎng)層凋亡細胞核呈棕黃色,部分核中有棕黃色顆粒。凋亡細胞核較正常增大,呈橢圓形或不規(guī)則形,核內(nèi)染色質(zhì)聚集于邊緣處。兩種凋亡檢測方法均顯示,鉛暴露組胎盤細胞凋亡指數(shù)高于對照組,且孕期全程鉛暴露組高于孕早期和孕晚期鉛暴露組(P0.01)。③Smad4主要表達于大鼠胎盤絨毛滋養(yǎng)細胞、毛細血管及間質(zhì)細胞,在實驗組中的表達較對照組增高,差異有統(tǒng)計學意義(F=10.08,P0.01)。與孕早期鉛暴露組比較,孕晚期、孕期全程鉛暴露組大鼠胎盤的Smad4表達下降,差異有統(tǒng)計學意義(P0.05)。④S100B蛋白主要表達于大鼠胎盤細胞滋養(yǎng)層細胞、蛻膜細胞及血管壁細胞胞質(zhì)中,實驗組陽性表達率高于對照組(52.94%),且孕期全程鉛暴露組(93.33%)高于孕早期(87.50%)和孕晚期鉛暴露組(80.00%),差異有統(tǒng)計學意義(x2=18.62,P0.05)。 結論胎盤細胞凋亡、Smad4及S100B蛋白在孕期不同階段鉛暴露下大鼠胎盤中的表達與孕末期血鉛水平密切相關。細胞凋亡是鉛致胎盤損傷的重要機制之一。Smad4及S100B蛋白在鉛致胎盤細胞凋亡的發(fā)生發(fā)展中具有重要作用。
[Abstract]:Objective to investigate the expression of Smad4 and S100B proteins in placenta of rats exposed to lead during pregnancy. Methods 108 rats were randomly divided into 4 groups. 0.025% lead acetate solution was taken at different stages of pregnancy. Determination of blood lead level. Hoechst nuclear staining by atomic absorption spectrometry and detection of apoptosis by deoxyribonucleotide terminal transferase-mediated Nick end labeling (Tunel) technique. The expression of Smad4 and S100B protein in placenta of rats exposed to lead in each group was determined by immunohistochemical method. The results were analyzed by single factor analysis of variance (ANOVA). The counting data were compared by chi-square test. Results (1) the blood lead levels in the experimental group were higher than 0.483 渭 mol / L, and there was a significant difference between the experimental group and the control group in the blood lead levels during different pregnancy periods. The results of Hoechst staining showed that the apoptotic nuclei of placental trophoblastic layer in the lead exposure group were strongly stained. Dense massive fluorescent granules could be seen in some nuclei. The apoptotic cells in the control group were scattered in distribution. Tunel technique showed that the apoptotic nuclei of placental trophoblastic layer of rats exposed to lead were brownish yellow. There were brown granules in some nuclei. The apoptotic nuclei were oval or irregular in shape. The chromatin in the nuclei gathered at the edge. The apoptotic index of placenta cells in lead exposure group was higher than that in control group, both methods showed that the apoptosis index of placental cells in lead exposure group was higher than that in control group. The results showed that the expression of P0.01C. 3Smad4 in placental villus trophoblast, capillary and interstitial cells was higher in the experimental group than in the control group, and it was higher in the lead exposure group during pregnancy than that in the early and late pregnancy groups, and the expression of Smad4 was higher in the experimental group than in the control group. Compared with the lead exposure group in early pregnancy, the expression of Smad4 in placenta of rats exposed to lead during pregnancy was decreased, and the protein of P0.05, 4S100B was mainly expressed in trophoblast cells of rat placenta. The positive expression rate of decidual cells and vascular wall cells in the experimental group was higher than that in the control group (52.94) and the lead exposure group during pregnancy (93.33) was higher than that in the first trimester of pregnancy (87.50%) and in the late stage of pregnancy (80.005%). The difference was statistically significant. Conclusion the expression of Smad4 and S100B proteins in placenta of rats exposed to lead in different stages of pregnancy is closely related to the level of blood lead in late pregnancy. Apoptosis is one of the important mechanisms of placental injury induced by lead, Smad4 and S100B proteins. Lead-induced placental apoptosis plays an important role in the development of placental cell apoptosis.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R179

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