本文關(guān)鍵詞： 氯乙烯單體 細(xì)胞周期 G1/S關(guān)卡 細(xì)胞周期蛋白依賴性激酶　出處：《山西醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 通過研究氯乙烯單體(vinyl chloride monomer, VCM)對(duì)大鼠肝細(xì)胞P53介導(dǎo)的細(xì)胞周期G1/S關(guān)卡的影響和相關(guān)蛋白表達(dá)的變化,以期探索VCM可能的致癌機(jī)制。 材料與方法 將64只SD大鼠隨機(jī)分成4組,分別為5、25、125mg/kg體重3個(gè)染毒組和對(duì)照組,對(duì)照組為同中劑量染毒組相同體積的清潔空氣,采用腹腔注射染毒,每周3次,持續(xù)12周。實(shí)驗(yàn)過程中記錄大鼠的一般情況。染毒6周和12周分別隨機(jī)處死32只大鼠(雌雄各半),流式細(xì)胞術(shù)檢測(cè)大鼠肝細(xì)胞周期和凋亡,Western blot檢測(cè)肝細(xì)胞周期相關(guān)蛋白(正性調(diào)控cyclinD1和負(fù)性調(diào)控P53、P21)的表達(dá),免疫組織化學(xué)法檢測(cè)P16蛋白表達(dá)的變化。所有數(shù)據(jù)均采用SPSS16.0統(tǒng)計(jì)軟件進(jìn)行分析。 結(jié)果 1.大鼠的一般狀況在染毒過程中大鼠未發(fā)現(xiàn)明顯中毒癥狀,體重正常增加,4組間差異沒有統(tǒng)計(jì)學(xué)意義(P0.05)。 2.大鼠染毒6周后,大鼠肝細(xì)胞G0/G1期分布差異P=0.055,各染毒組的G0/G1期分布百分比均高于對(duì)照組。染毒12周時(shí),S期細(xì)胞百分比隨著染毒劑量的增加而增加(P=0.016)。 3.VCM對(duì)大鼠染毒6周后,大鼠肝細(xì)胞P21蛋白表達(dá)量隨染毒劑量的增加而降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05),P53蛋白表達(dá)量隨染毒劑量的增加而增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05), cyclinA和cyclinD1蛋白表達(dá)量均隨染毒劑量的增加而增加,差異均有統(tǒng)計(jì)學(xué)意義(P0.001)。 4.VCM對(duì)大鼠染毒12周后,P21蛋白的表達(dá)量隨染毒劑量的增加而增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05),中、高劑量組P21蛋白表達(dá)量高于對(duì)照組,高劑量組高于低劑量組,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)；P53蛋白的表達(dá)量隨染毒劑量的增加呈上升趨勢(shì),高劑量組高于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05),但總體差異沒有統(tǒng)計(jì)學(xué)意義(P0.05)。cyclinA和cyclinD1蛋白表達(dá)量隨染毒劑量增加而增加,差異均有統(tǒng)計(jì)學(xué)意義(P0.05),其中高劑量組表達(dá)量高于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.001)。 5.免疫組化法檢測(cè)VCM染毒12周,P16蛋白表達(dá)量各組間差異沒有統(tǒng)計(jì)學(xué)意義(P0.05),CDK4蛋白表達(dá)量隨染毒劑量的增加呈降低趨勢(shì),染毒組的蛋白表達(dá)量均較對(duì)照組降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 6.組織病理學(xué)檢查發(fā)現(xiàn)VCM染毒后,對(duì)照組和低劑量組肝細(xì)胞形態(tài)基本無變化,中劑量和高劑量組肝細(xì)胞有氣球樣變,匯管區(qū)變大,纖維化等病理改變。 結(jié)論 1.VCM在低劑量早期暴露時(shí),能夠影響細(xì)胞周期分布,引起G0/G1期阻滯,隨著染毒劑量的增加和染毒時(shí)間的延長G1期阻滯消失,發(fā)生S期阻滯。 2.VCM染毒對(duì)大鼠肝細(xì)胞早期凋亡有影響,對(duì)晚期凋亡的影響不大。 3. cyclin A、cyclinD1和P21蛋白的過表達(dá)可能與氯乙烯導(dǎo)致的細(xì)胞增殖失控有關(guān)。
[Abstract]:Purpose Vinyl chloride monomer (VCM) vinyl chloride monomer was studied. The effect of VCM on p53 mediated G1 / S level and the expression of related proteins in rat hepatocytes were studied in order to explore the possible carcinogenic mechanism of VCM. Materials and methods 64 Sprague-Dawley rats were randomly divided into 4 groups: the control group and the control group. The rats in the control group were given the same volume of clean air at the same dose. The rats were injected intraperitoneally three times a week for 12 weeks. During the experiment, 32 rats (half of male and half female) were randomly killed at 6 and 12 weeks of exposure. Flow cytometry was used to detect hepatocyte cycle and apoptosis. Western blot was used to detect hepatocyte cycle associated protein (positive regulation of cyclinD1 and negative regulation of p53). The expression of P21) and the expression of P16 protein were detected by immunohistochemistry. All the data were analyzed by SPSS16.0 software. Results 1. During the course of exposure, no obvious toxic symptoms were found in rats, and there was no significant difference among the four groups in normal weight gain (P 0.05). 2. After 6 weeks of exposure, the difference of G0 / G1 phase distribution in rat hepatocytes was 0.055. The percentage of G0 / G1 phase distribution in each exposure group was higher than that in the control group. At 12 weeks of exposure, the percentage of G0 / G1 phase distribution was higher than that of the control group. The percentage of S phase cells increased with the increase of exposure dose. 3. After 6 weeks of VCM exposure, the expression of P21 protein decreased with the increase of the dose of VCM, and the difference was statistically significant (P 0.05). The expression of p53 protein increased with the increase of exposure dose, the difference was statistically significant (P0.05). The expression of cyclinA and cyclinD1 protein increased with the increase of exposure dose, and the difference was statistically significant (P 0.001). 4. After 12 weeks of VCM exposure, the expression of P21 protein increased with the increase of the dose of VCM, and the difference was statistically significant (P 0.05). The expression of P21 protein in the high dose group was higher than that in the control group, and in the high dose group was higher than that in the low dose group (P 0.05). The expression of p53 protein in high dose group was higher than that in control group, the difference was statistically significant (P 0.05). But the total difference was not statistically significant (P0.05A. Cyclin A and cyclinD1 protein expression increased with the increase of exposure dose, the difference was statistically significant (P0.05). The expression level of high dose group was higher than that of control group, the difference was statistically significant (P 0.001). 5. There was no significant difference in the expression of P16 protein between the groups exposed to VCM for 12 weeks by immunohistochemical method. The expression of CDK4 protein decreased with the increase of the dose of VCM. The expression of protein in the exposed group was lower than that in the control group, and the difference was statistically significant (P 0.05). 6. Histopathological examination showed that the morphology of hepatocytes in the control group and the low dose group was almost unchanged after VCM exposure, and the hepatic cells in the middle and high dose groups had balloon changes and the catchment area became larger. Fibrosis and other pathological changes. Conclusion 1. VCM could affect the cell cycle distribution and induce G _ 0 / G _ 1 phase arrest during early exposure at low dose, which disappeared with the increase of exposure dose and the prolongation of exposure time. Stage S block occurred. 2. VCM exposure had an effect on early apoptosis of rat hepatocytes, but had little effect on late apoptosis. 3. Overexpression of cyclin cyclin D1 and P21 proteins may be related to uncontrolled cell proliferation induced by vinyl chloride.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 任雪峰;氯乙烯的基因毒性[J];國外醫(yī)學(xué)(衛(wèi)生學(xué)分冊(cè));2000年04期

2 吳維皚,李霜;氯乙烯仍受關(guān)注的原因[J];國外醫(yī)學(xué)(衛(wèi)生學(xué)分冊(cè));2001年03期

3 王民生,蔣曉紅,王湘蘇,徐德洲,P．Schmezer,R．G．Klein;氯乙烯致大鼠肝細(xì)胞和外周血淋巴細(xì)胞DNA損傷的活體實(shí)驗(yàn)研究[J];工業(yè)衛(wèi)生與職業(yè)病;1998年04期

4 王民生,翟明芬,張若陽,王湘蘇,徐德州;用單細(xì)胞微量凝膠電泳檢測(cè)氯乙烯暴露工人DNA損傷的研究[J];癌變.畸變.突變;1999年06期

5 劉俊玲,保毓書;氯乙烯對(duì)女工生殖健康的影響及其預(yù)防的研究[J];河南預(yù)防醫(yī)學(xué)雜志;1997年04期

6 葉菲;金銀姬;金曉明;王天真;;周期蛋白A、E在胃低分化腺癌中表達(dá)的研究[J];哈爾濱醫(yī)科大學(xué)學(xué)報(bào);2006年01期

7 吳小明;p73基因與p53抑癌基因的比較[J];惠州學(xué)院學(xué)報(bào)(自然科學(xué)版);2003年06期

8 王民生;蔣曉紅;常元?jiǎng)?;氯乙烯致癌作用與危險(xiǎn)度評(píng)價(jià)[J];江蘇預(yù)防醫(yī)學(xué);2012年02期

9 朱爭(zhēng)艷,杜智,孫銘,高英堂,齊之麗;p16真核表達(dá)載體的構(gòu)建及其對(duì)肝癌細(xì)胞的作用[J];臨床肝膽病雜志;2003年01期

10 任雪峰,柴尚健,萬俊香,帕爾哈提,吳翠娥,夏昭林,David Christiani;氯乙烯作業(yè)工人職業(yè)接觸評(píng)定與健康危害調(diào)查[J];勞動(dòng)醫(yī)學(xué);2001年01期

，

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