自噬基因LC-3與子宮內(nèi)膜異位癥的相關(guān)性研究
發(fā)布時(shí)間：2018-08-08 21:20
【摘要】：研究背景： 子宮內(nèi)膜異位癥（Endometriosis, EMs）是一種常見(jiàn)和多發(fā)的婦科內(nèi)分泌疾病，其引起的不孕及慢性疼痛癥狀嚴(yán)重影響育齡婦女生殖健康和生活質(zhì)量。目前EMs的發(fā)病機(jī)制不明。 程序性細(xì)胞死亡(Programmed ce1l death，，PCD)由多種信號(hào)通路控制并維持細(xì)胞內(nèi)環(huán)境的穩(wěn)定性。PCD分為兩種：細(xì)胞凋亡(屬I(mǎi)型細(xì)胞死亡，由caspase介導(dǎo))和自體吞噬(屬I(mǎi)I型細(xì)胞死亡，非caspase介導(dǎo))。自噬與溶酶體相融合形成自噬溶酶體將吞噬物降解消化，吞噬物包括蛋白質(zhì)及細(xì)胞器等。自噬在多種疾病的發(fā)生發(fā)展中發(fā)揮著作用。 微管相關(guān)輕鏈蛋白-3（Microtubule associated protein light chain3，LC-3）是酵母細(xì)胞自噬相關(guān)基因ATG8的同源基因。LC-3在多種腫瘤疾病中的表達(dá)水平出現(xiàn)異常，而子宮內(nèi)膜異位癥與腫瘤又有相似的生物學(xué)行為，目前細(xì)胞自噬與子宮內(nèi)膜異位癥發(fā)病機(jī)制關(guān)系的研究報(bào)道較少，本實(shí)驗(yàn)對(duì)細(xì)胞自噬泡超微結(jié)構(gòu)及自噬基因LC-3與子宮內(nèi)膜異位癥發(fā)病機(jī)制進(jìn)行相關(guān)性研究。 目的： 探討子宮內(nèi)膜異位癥患者在位內(nèi)膜及異位內(nèi)膜細(xì)胞自噬泡超微結(jié)構(gòu)變化；LC-3mRNA和蛋白在子宮內(nèi)膜異位癥在位內(nèi)膜和異位內(nèi)膜細(xì)胞中的表達(dá)及意義。 方法： 本文收集31例內(nèi)異癥患者的異位內(nèi)膜組織及該患者相應(yīng)的在位內(nèi)膜組織為實(shí)驗(yàn)組，31例正常的子宮內(nèi)膜組織為對(duì)照組，應(yīng)用透射電鏡觀察內(nèi)異癥在位及異位內(nèi)膜中細(xì)胞自噬泡超微結(jié)構(gòu)的改變；RT-PCR、Western-blotting方法檢測(cè)LC-3mRNA和蛋白在內(nèi)異癥組在位內(nèi)膜細(xì)胞、異位內(nèi)膜細(xì)胞及正常子宮內(nèi)膜細(xì)胞中的表達(dá)，利用SPSS19.0統(tǒng)計(jì)學(xué)軟件分析實(shí)驗(yàn)結(jié)果。 結(jié)果 實(shí)驗(yàn)組和對(duì)照組患者的平均年齡分別為：(32.48±4.36)歲；(32.39±2.64)歲，兩組間比較無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05);透射電鏡結(jié)果分析顯示：實(shí)驗(yàn)組的在位內(nèi)膜細(xì)胞及異位內(nèi)膜細(xì)胞中的自噬泡數(shù)目較對(duì)照組的正常子宮內(nèi)膜細(xì)胞的自噬泡數(shù)目有所下調(diào)；Real time-PCR結(jié)果分析顯示：LC-3mRNA在實(shí)驗(yàn)組的在位子宮內(nèi)膜細(xì)胞及異位內(nèi)膜細(xì)胞中表達(dá)均低于對(duì)照組內(nèi)膜細(xì)胞(P=0.015)，實(shí)驗(yàn)組在位子宮內(nèi)膜細(xì)胞的LC-3mRNA較對(duì)照組內(nèi)膜細(xì)胞是下調(diào)的（0.435±0.227; P=0.013），實(shí)驗(yàn)組異位子宮內(nèi)膜細(xì)胞的LC-3mRNA表達(dá)量比對(duì)照組降低(0.429±0.303; P=0.011)，但實(shí)驗(yàn)組在位內(nèi)膜及異位內(nèi)膜細(xì)胞的LC-3mRNA表達(dá)量無(wú)明顯差異（1.282±0.534;P=0.949）；Western Blot結(jié)果分析顯示：LC-3蛋白在實(shí)驗(yàn)組的在位子宮內(nèi)膜細(xì)胞及異位內(nèi)膜細(xì)胞中表達(dá)均低于對(duì)照組內(nèi)膜(P=0.000)，實(shí)驗(yàn)組在位子宮內(nèi)膜細(xì)胞的LC-3蛋白較對(duì)照組內(nèi)膜細(xì)胞是下調(diào)的（0.242±0.08，0.440±0.08；P=0.000）,實(shí)驗(yàn)組異位子宮內(nèi)膜細(xì)胞的LC-3蛋白表達(dá)量比對(duì)照組降低(0.309±0.09；P=0.000)，但同時(shí)實(shí)驗(yàn)組異位內(nèi)膜細(xì)胞的LC-3蛋白表達(dá)量明顯高于在位內(nèi)膜組（P=0.003）。 結(jié)論 子宮內(nèi)膜異位癥患者的異位內(nèi)膜細(xì)胞的自噬活性降低，同時(shí)在位內(nèi)膜細(xì)胞的自噬活性也降低，自噬參與了子宮內(nèi)膜異位癥的發(fā)病。
[Abstract]:Research background:
Endometriosis (EMs) is a common and multiple gynecologic endocrine disease. The symptoms of infertility and chronic pain seriously affect the reproductive health and quality of life of women of childbearing age. The pathogenesis of EMs is unclear.
Programmed ce1l death (PCD) is controlled by a variety of signaling pathways and maintains the stability of the intracellular environment,.PCD is divided into two kinds: apoptosis (I cell death, caspase mediated) and autophagy (II cell death, non caspase mediating). Autophagy and lysosomes are fused to form autophagic lysosomes to reduce phagocytosis. Digestion and phagocytosis include proteins and organelles. Autophagy plays a role in the development of many diseases.
Microtubule related light chain protein -3 (Microtubule associated protein light chain3, LC-3) is the expression level of the homologous gene.LC-3 in yeast cell autophagy related gene ATG8 in a variety of tumor diseases, and endometriosis and tumor have similar biological behavior. At present, the pathogenesis of autophagy and endometriosis is the disease. There are few reports on the mechanism of endometriosis. In this study, the ultrastructure of autophagic vesicles and the correlation between autophagic gene LC-3 and the pathogenesis of endometriosis were studied.
Objective:
To investigate the ultrastructural changes of autophagic vesicles in eutopic and ectopic endometrium cells of endometriosis, and the expression and significance of LC-3mRNA and protein in eutopic and ectopic endometrium cells of endometriosis.
Method:
The ectopic endometrium of 31 patients with endometriosis and the corresponding eutopic endometrium in the experimental group were collected and 31 normal endometrium tissues were used as the control group. The ultrastructural changes of the autophagic vesicles in the eutopic and ectopic endometrium were observed by transmission electron microscopy, and the RT-PCR and Western-blotting methods were used to detect LC-3mRNA and protein. In endometriosis group, the expression of eutopic endometrial cells, ectopic endometrial cells and normal endometrial cells was analyzed by SPSS19.0 statistical software.
Result
The average age of the patients in the experimental group and the control group was (32.48 + 4.36) years and (32.39 + 2.64) years old, and there was no statistical significance between the two groups (P > 0.05). The number of autophagic vacuoles in the eutopic and ectopic endometrium cells of the experimental group was compared with the number of autophagic vesicles in the normal endometrium cells of the control group. The results of Real time-PCR analysis showed that the expression of LC-3mRNA in the eutopic endometrium and ectopic endometrium cells in the experimental group was lower than that of the control group (P=0.015). The LC-3mRNA of the eutopic endometrium cells in the experimental group was lower than that of the control group (0.435 + 0.227; P=0.013), and the endometrium endometrium was fine in the experimental group. The expression of LC-3mRNA in the cell was lower than that of the control group (0.429 + 0.303; P=0.011), but the expression of LC-3mRNA in the eutopic and ectopic endometrium cells of the experimental group was not significantly different (1.282 + 0.534; P=0.949). The analysis of Western Blot results showed that the expression of LC-3 protein in the eutopic and ectopic endometrium cells of the experimental group was lower than that of the control. The LC-3 protein of the eutopic endometrium cells in the experimental group was lower than that of the control group (0.242 + 0.08,0.440 + 0.08; P=0.000). The expression of LC-3 protein in the ectopic endometrium cells in the experimental group was lower than that of the control group (0.309 + 0.09; P=0.000), but the expression of LC-3 protein in the ectopic endometrium cells in the experimental group was obvious at the same time. Higher than the eutopic endometrium group (P=0.003).
conclusion
The autophagy activity of ectopic endometrium cells in patients with endometriosis is reduced and autophagy of the eutopic cells is also reduced, and autophagy is involved in the pathogenesis of endometriosis.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R711.71
本文編號(hào)：2173072
[Abstract]:Research background:
Endometriosis (EMs) is a common and multiple gynecologic endocrine disease. The symptoms of infertility and chronic pain seriously affect the reproductive health and quality of life of women of childbearing age. The pathogenesis of EMs is unclear.
Programmed ce1l death (PCD) is controlled by a variety of signaling pathways and maintains the stability of the intracellular environment,.PCD is divided into two kinds: apoptosis (I cell death, caspase mediated) and autophagy (II cell death, non caspase mediating). Autophagy and lysosomes are fused to form autophagic lysosomes to reduce phagocytosis. Digestion and phagocytosis include proteins and organelles. Autophagy plays a role in the development of many diseases.
Microtubule related light chain protein -3 (Microtubule associated protein light chain3, LC-3) is the expression level of the homologous gene.LC-3 in yeast cell autophagy related gene ATG8 in a variety of tumor diseases, and endometriosis and tumor have similar biological behavior. At present, the pathogenesis of autophagy and endometriosis is the disease. There are few reports on the mechanism of endometriosis. In this study, the ultrastructure of autophagic vesicles and the correlation between autophagic gene LC-3 and the pathogenesis of endometriosis were studied.
Objective:
To investigate the ultrastructural changes of autophagic vesicles in eutopic and ectopic endometrium cells of endometriosis, and the expression and significance of LC-3mRNA and protein in eutopic and ectopic endometrium cells of endometriosis.
Method:
The ectopic endometrium of 31 patients with endometriosis and the corresponding eutopic endometrium in the experimental group were collected and 31 normal endometrium tissues were used as the control group. The ultrastructural changes of the autophagic vesicles in the eutopic and ectopic endometrium were observed by transmission electron microscopy, and the RT-PCR and Western-blotting methods were used to detect LC-3mRNA and protein. In endometriosis group, the expression of eutopic endometrial cells, ectopic endometrial cells and normal endometrial cells was analyzed by SPSS19.0 statistical software.
Result
The average age of the patients in the experimental group and the control group was (32.48 + 4.36) years and (32.39 + 2.64) years old, and there was no statistical significance between the two groups (P > 0.05). The number of autophagic vacuoles in the eutopic and ectopic endometrium cells of the experimental group was compared with the number of autophagic vesicles in the normal endometrium cells of the control group. The results of Real time-PCR analysis showed that the expression of LC-3mRNA in the eutopic endometrium and ectopic endometrium cells in the experimental group was lower than that of the control group (P=0.015). The LC-3mRNA of the eutopic endometrium cells in the experimental group was lower than that of the control group (0.435 + 0.227; P=0.013), and the endometrium endometrium was fine in the experimental group. The expression of LC-3mRNA in the cell was lower than that of the control group (0.429 + 0.303; P=0.011), but the expression of LC-3mRNA in the eutopic and ectopic endometrium cells of the experimental group was not significantly different (1.282 + 0.534; P=0.949). The analysis of Western Blot results showed that the expression of LC-3 protein in the eutopic and ectopic endometrium cells of the experimental group was lower than that of the control. The LC-3 protein of the eutopic endometrium cells in the experimental group was lower than that of the control group (0.242 + 0.08,0.440 + 0.08; P=0.000). The expression of LC-3 protein in the ectopic endometrium cells in the experimental group was lower than that of the control group (0.309 + 0.09; P=0.000), but the expression of LC-3 protein in the ectopic endometrium cells in the experimental group was obvious at the same time. Higher than the eutopic endometrium group (P=0.003).
conclusion
The autophagy activity of ectopic endometrium cells in patients with endometriosis is reduced and autophagy of the eutopic cells is also reduced, and autophagy is involved in the pathogenesis of endometriosis.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R711.71
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