本文選題：核轉(zhuǎn)錄因子kappa + B��； 參考：《中南大學(xué)》2014年碩士論文

【摘要】：目的： 1、探討NMB介導(dǎo)的NF-κB及AP-1的表達(dá)量變化和相關(guān)性與分娩啟動(dòng)的關(guān)系。 2、研究子宮下段平滑肌細(xì)胞中NMB介導(dǎo)的NF-κB及AP-1發(fā)生細(xì)胞內(nèi)易位與分娩啟動(dòng)的關(guān)系。 方法： 1、組織塊貼壁法原代培養(yǎng)人妊娠晚期子宮下段平滑肌細(xì)胞,采用免疫細(xì)胞化學(xué)方法進(jìn)行鑒定。 2、臨產(chǎn)前后子宮下段平滑肌組織采用石蠟包埋固定,采用蘇木精-伊紅染色方法進(jìn)行切片質(zhì)量檢測(cè)。 3、采用免疫組織化學(xué)方法對(duì)自然臨產(chǎn)前后子宮下段平滑肌細(xì)胞中NF-κB及AP-1表達(dá)進(jìn)行測(cè)定。 4、采用免疫細(xì)胞熒光對(duì)NMB誘導(dǎo)前后子宮下段平滑肌細(xì)胞中NF-κB(p65)及AP-1(c-Jun)表達(dá)進(jìn)行測(cè)定,并通過(guò)多重免疫熒光染色進(jìn)行NF-κB及AP-1的細(xì)胞內(nèi)共定位。 5、采用Image-Pro Plus6.0軟件進(jìn)行NF-κB及AP-1表達(dá)的累積光密度值(IOD)測(cè)定,采用SPSS19.0軟件分析NF-κB及AP-1表達(dá)的差異及相關(guān)性。 結(jié)果： 1、組織塊貼壁法培養(yǎng)的原代子宮平滑肌細(xì)胞生長(zhǎng)狀態(tài)良好,經(jīng)免疫細(xì)胞化學(xué)法α-SMA鑒定及NMBR檢測(cè),細(xì)胞純度近100%,說(shuō)明培養(yǎng)的細(xì)胞是表達(dá)NMBR的子宮平滑肌細(xì)胞。 2、組織經(jīng)石蠟包埋固定,行HE染色檢測(cè)。子宮下段平滑肌細(xì)胞結(jié)構(gòu)清晰,說(shuō)明組織固定包埋方法可行,組織切片適于進(jìn)行免疫組織化學(xué)染色檢測(cè)。 3、自然臨產(chǎn)前后子宮下段平滑肌細(xì)胞中p65、c-Jun定位：未臨產(chǎn)組p65與c-Jun主要位于細(xì)胞漿,臨產(chǎn)組p65與c-Jun主要位于細(xì)胞核。并且臨產(chǎn)組與未臨產(chǎn)組p65、c-Jun表達(dá)量存在差異,臨產(chǎn)組p65及c-Jun表達(dá)量高于未臨產(chǎn)組,差異具有統(tǒng)計(jì)學(xué)意義,P0.05。 4、自然臨產(chǎn)前后子宮平滑肌細(xì)胞中p65與c-Jun表達(dá)量Pearson相關(guān)性分析：未臨產(chǎn)組及臨床組的p65及c-Jun表達(dá)均存在正相關(guān),相關(guān)系數(shù)分別為0.659、0.879,p0.05。 5、NMB誘導(dǎo)前后p65、c-Jun定位：未加NMB的對(duì)照組位于細(xì)胞漿；NMB4h組主要位于核膜周邊區(qū)域；NMB8h組主要位于細(xì)胞核,細(xì)胞漿呈低表達(dá)。 6、NMB誘導(dǎo)前后的p65、c-Jun表達(dá)：NMB8h組IOD值明顯高于對(duì)照組及NMB4h組,差異具有統(tǒng)計(jì)學(xué)意義,P0.05;NMB4h組與對(duì)照組之間差異無(wú)統(tǒng)計(jì)學(xué)意義,p0.05。 7、p65與c-Jun共定位：對(duì)照組、NMB4h組、NMB8h組,多重免疫熒光染色顯示二者在細(xì)胞內(nèi)表達(dá)部位一致,NMB誘導(dǎo)前二者主要位于細(xì)胞漿,NMB誘導(dǎo)后8小時(shí)二者主要位于細(xì)胞核,表達(dá)部位幾乎完全重疊。 8、NMB誘導(dǎo)的p65與c-Jun表達(dá)量Pearson相關(guān)性分析：對(duì)照組、NMB4h組、NMB8h組中,p65與c-Jun均呈正相關(guān),相關(guān)系數(shù)分別為0.587、0.653、0.874,P0.05。 結(jié)論： 1.分娩啟動(dòng)前,NF-κB及AP-1定位于細(xì)胞漿,分娩啟動(dòng)后NF-κB及AP-1遷移進(jìn)入細(xì)胞核。 2.NMB可同時(shí)誘導(dǎo)NF-κB及AP-1易位,二者定位及遷移方向呈同步變化,表達(dá)量同時(shí)增高,推測(cè)NF-κB與AP-1可能存在協(xié)同作用。
[Abstract]:Objective: 1. To investigate the relationship between the expression of NF- 魏 B and AP-1 mediated by NMB and the onset of labor. 2. To study the relationship between the intracellular translocation of NF- 魏 B and AP-1 mediated by NMB and the initiation of labor in the lower uterine smooth muscle cells. Methods: 1. The primary culture of human lower uterine smooth muscle cells in late pregnancy was identified by immunocytochemistry. 2. The smooth muscle tissue of the lower segment of uterus was fixed with paraffin and stained with hematoxylin-eosin before and after labor. 3. The expression of NF- 魏 B and AP-1 in the smooth muscle cells of the lower segment of uterus before and after spontaneous labor was determined by immunohistochemical method. (4) the expression of NF- 魏 B p65) and AP-1Tc-Jun) in the smooth muscle cells of the lower uterine segment before and after NMB induction were detected by immunofluorescence, and the intracellular localization of NF- 魏 B and AP-1 were detected by multiple immunofluorescence staining. 5. The cumulative optical density (IOD) of NF- 魏 B and AP-1 expression was determined by Image-Pro Plus6.0 software, and the difference and correlation of NF- 魏 B and AP-1 expression were analyzed by SPSS19.0 software. Results: 1. The primary uterine smooth muscle cells cultured by tissue mass adherence method grew well. The purity of the cells was nearly 100 by immunocytochemistry and NMBR detection, indicating that the cultured cells were uterine smooth muscle cells expressing NMBR. 2. The tissues were fixed with paraffin and stained with HE. The clear structure of smooth muscle cells in the lower segment of uterus showed that the method of tissue fixation and embedding was feasible and the tissue sections were suitable for immunohistochemical staining. 3. The localization of p65 c-Jun in the smooth muscle cells of the lower segment of uterus before and after spontaneous labor: p65 and c-Jun were mainly located in the cytoplasm in the non-parturient group, and p65 and c-Jun in the nucleus in the parturient group. The expression of p65 c-Jun in parturient group was higher than that in non-parturient group, and the difference was statistically significant (P 0.05). 4. Pearson correlation analysis of p65 and c-Jun expression in uterine smooth muscle cells before and after spontaneous labor: there was a positive correlation between p65 and c-Jun expression in unpartum group and clinical group, the correlation coefficient was 0.659 ~ 0.879p 0.05, respectively. Before and after induction of NMB, the position of c-Jun in the control group without NMB was mainly located in the nucleus in the peripheral region of the nuclear membrane, and the cytoplasm of the control group was low in the cytoplasm of NMB4 h group. The expression of p65 c-Jun was significantly higher in the 8h group than that in the control group and NMB4h group before and after induction, and there was no significant difference between the control group and the control group (P 0.05). 7p65 and c-Jun co-localization: in the control group of 4 h after NMB induction, multiple immunofluorescence staining showed that both of them were mainly located in the nucleus 8 hours after induction of NMB in cytoplasm. The site of expression was almost completely overlapped. Analysis of Pearson correlation between p65 and c-Jun expression induced by NMB: there was a positive correlation between p65 and c-Jun in control group (n = 4), and the correlation coefficient was 0.5870.653U 0.874P 0.05, respectively. Conclusion: 1. NF- 魏 B and AP-1 were located in cytoplasm before parturition, and NF- 魏 B and AP-1 migrated into nucleus after parturition. 2.NMB could induce the translocation of NF- 魏 B and AP-1 at the same time. The localization and migration direction of NF- 魏 B and AP-1 were changed synchronously, and the expression of NF- 魏 B was increased simultaneously. It is speculated that NF- 魏 B and AP-1 might have synergistic effect.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R714

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相關(guān)期刊論文 前1條

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本文編號(hào)：1971894