發(fā)布時(shí)間：2018-05-06 10:07

  本文選題：子癇前期 + 細(xì)胞因子信號(hào)轉(zhuǎn)導(dǎo)蛋白抑制因子��； 參考：《南京醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的： 探究細(xì)胞因子信號(hào)轉(zhuǎn)導(dǎo)抑制因子3(suppressor of cytokine signaling3, SOCS3)基因在人子癇前期胎盤組織中的表達(dá)情況及其對(duì)HTR-8/SVneo細(xì)胞增殖與遷移能力的影響。 方法： 1.選取2011年10月至2012年10月在南京醫(yī)科大學(xué)第一附屬醫(yī)院住院分娩的15例重度子癇前期孕婦為子癇前期組,15例正常孕婦作為正常妊娠組。 2.采用實(shí)時(shí)熒光定量逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)和Western印跡技術(shù)檢測(cè)兩組孕婦胎盤組織中SOCS3mRNA和蛋白表達(dá)水平。 3.體外培養(yǎng)的HTR-8/SVneo細(xì)胞分別轉(zhuǎn)染SOCS3小分子干擾RNA(實(shí)驗(yàn)組)和無意義的陰性對(duì)照小分子干擾RNA(陰性對(duì)照組)。4.采用實(shí)時(shí)熒光定量逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)和Western印跡技術(shù)檢測(cè)體外培養(yǎng)的HTR-8/SVneo細(xì)胞中SOCS3mRNA和蛋白表達(dá)水平。 5.采用克隆形成實(shí)驗(yàn)和3-(4,5)-二甲基噻唑-(2,5)-二苯基溴化四氮唑藍(lán)法(MTT)檢測(cè)轉(zhuǎn)染后兩個(gè)實(shí)驗(yàn)組中細(xì)胞的增殖能力。 6.應(yīng)用流式細(xì)胞儀對(duì)轉(zhuǎn)染后兩個(gè)實(shí)驗(yàn)組中細(xì)胞進(jìn)行細(xì)胞周期分析。 7.采用transwell小室實(shí)驗(yàn)檢測(cè)轉(zhuǎn)染后兩個(gè)實(shí)驗(yàn)組中細(xì)胞遷移能力。 結(jié)果： 1.子癇前期組的胎盤組織中SOCS3mRNA和SOCS3蛋白水平顯著低于正常組胎盤組織,分別為0.254±0.03和0.21±0.05,均低于正常妊娠組0.71±0.08和0.75±0.12。差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。 2.小分子干擾RNA轉(zhuǎn)染24h后,實(shí)驗(yàn)組SOCS3mRNA水平低于陰性對(duì)照組0.39±0.02與1.0±0.04,蛋白水平也較低(0.0037+0.0014與1.5149±0.0357)。實(shí)驗(yàn)組中SOCS3mRNA和SOCS3蛋白水平表達(dá)均降低,差異均有統(tǒng)計(jì)學(xué)意義(P0.05),證明干擾SOCS3表達(dá)成功。 3.MTT法檢測(cè)證實(shí)實(shí)驗(yàn)組細(xì)胞增殖能力降低,轉(zhuǎn)染后48、72和96h實(shí)驗(yàn)組增殖能力分別為0.23±0.01,0.32±0.02和0.3±0.02,均低于陰性對(duì)照組(分別為0.39±0.02,0.55±0.04和0.86±0.04),差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 4.克隆形成實(shí)驗(yàn)證實(shí)轉(zhuǎn)染10d時(shí)實(shí)驗(yàn)組克隆形成細(xì)胞個(gè)數(shù)顯著低于陰性對(duì)照組,實(shí)驗(yàn)組細(xì)胞數(shù)為116±15,低于陰性對(duì)照組312±24,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 5.應(yīng)用流式細(xì)胞儀分析細(xì)胞周期發(fā)現(xiàn)轉(zhuǎn)染48h后,實(shí)驗(yàn)組G1/Go期細(xì)胞比例為(55.75±2.21)%,高于陰性對(duì)照組[(47.88±1.87)%](t＝45.43,P0.05)；S期細(xì)胞比例為(31.59±0.83)%,低于陰性對(duì)照組[(37.38±1.34)%](t=20.06,P0.05)。兩組分別比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。 6.transwell小室實(shí)驗(yàn)證實(shí)轉(zhuǎn)染48h后,實(shí)驗(yàn)組穿膜細(xì)胞數(shù)為(93±11)個(gè),低于陰性對(duì)照組(167±17)個(gè),兩組相比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論： 1.子癇前期患者胎盤組織中SOCS3水平與正常妊娠孕婦相比較表達(dá)下降,推測(cè)SOCS3可能參與了子癇前期的發(fā)生發(fā)展過程。 2. SOCS3可能通過降低表達(dá)水平來抑制滋養(yǎng)細(xì)胞的增殖和遷移能力,從而在子癇前期的發(fā)病過程中起作用。
[Abstract]:Objective: To investigate the expression of cytokine signal transduction inhibitor 3(suppressor of cytokine signaling3 (SOCS3) gene in human preeclampsia placenta and its effect on the proliferation and migration of HTR-8/SVneo cells. Methods: 1. From October 2011 to October 2012, 15 pregnant women with severe preeclampsia were selected as normal pregnancy group and 15 normal pregnant women in the first affiliated Hospital of Nanjing Medical University. 2. The expression of SOCS3mRNA and protein in placenta of pregnant women was detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction and Western blotting. 3. HTR-8/SVneo cells were transfected with SOCS3 small molecule interference RNAs (experimental group) and meaningless negative control group (negative control group) respectively. The expression of SOCS3mRNA and protein in cultured HTR-8/SVneo cells was detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction and Western blotting. 5. Clone formation assay and MTT assay were used to detect the proliferation of the cells in the two experimental groups after transfection. 6. Flow cytometry was used to analyze the cell cycle in the two experimental groups after transfection. 7. The ability of cell migration in the two experimental groups after transfection was detected by transwell chamber experiment. Results: 1. The levels of SOCS3mRNA and SOCS3 protein in placenta of preeclampsia group were significantly lower than those of normal group (0.254 鹵0.03 and 0.21 鹵0.05, respectively), and were lower than those of normal pregnancy group (0.71 鹵0.08) and 0.75 鹵0.12 (P < 0.05). The difference was statistically significant (P 0.05). 2. 24 hours after transfection with small interfering RNA, the level of SOCS3mRNA in the experimental group was lower than that in the negative control group (0.39 鹵0.02 vs 1.0 鹵0.04), and the protein level was also lower than that in the negative control group (0.0037 0.0014 and 1.5149 鹵0.0357). The expression of SOCS3mRNA and SOCS3 protein decreased in the experimental group, and the difference was statistically significant (P 0.05), which proved the interference of SOCS3 expression was successful. The proliferative ability of the experimental group was 0.23 鹵0.01 鹵0.032 鹵0.02 and 0.3 鹵0.02, respectively, which was significantly lower than that of the negative control group (0.39 鹵0.02 鹵0.55 鹵0.04 and 0.86 鹵0.04) at 48 ~ 72 and 96 h after transfection, respectively (P 0.05). 4. The number of Clone forming cells in the experimental group was significantly lower than that in the negative control group at 10 days after transfection, and the number of the cells in the experimental group was 116 鹵15, which was lower than that in the negative control group (312 鹵24). The difference was statistically significant (P 0.05). 5. Flow cytometry analysis showed that the percentage of G1/Go phase cells in the experimental group was 55.75 鹵2.21 after 48 h transfection, which was higher than that in the negative control group [47.88 鹵1.87%] TX 45.43 P 0.05%, and was lower than that in the negative control group (37.38 鹵1.34%), and the ratio of the cells in the S phase was 31.59 鹵0.83%, which was lower than that in the negative control group (37.38 鹵1.34%). The difference between the two groups was statistically significant (P 0.05). 6.transwell chamber experiment confirmed that the number of perforated cells in the experimental group was 93 鹵11, which was lower than that in the negative control group (167 鹵17) after 48 hours of transfection. The difference between the two groups was statistically significant (P 0.05). Conclusion: 1. The expression of SOCS3 in placenta of preeclampsia patients was lower than that of normal pregnant women. It was speculated that SOCS3 might be involved in the occurrence and development of preeclampsia. 2. SOCS3 may play a role in the pathogenesis of preeclampsia by inhibiting the proliferation and migration of trophoblast.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R714.2

【共引文獻(xiàn)】

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4 李，

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