本文選題：STAT3 + VEGF��； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:滋養(yǎng)細(xì)胞表達(dá)異�？赡苡绊懱ケP功能,引發(fā)妊娠相關(guān)疾病。本研究通過檢測在Poly(I:C)誘導(dǎo)下抑制人絨毛滋養(yǎng)層細(xì)胞HTR8-SVneo細(xì)胞中STAT3通路后VEGF、s Flt-1表達(dá)的變化,了解STAT3通路對滋養(yǎng)細(xì)胞VEGF、s Flt-1表達(dá)的影響。方法:1 Poly(I:C)刺激HTR8-SVneo細(xì)胞,收集刺激0h、2h、4h、6h、12h、24后的細(xì)胞,RT-PCR檢測s Flt-1,VEGF mRNA表達(dá)變化;2實(shí)驗(yàn)分為五組:(1)HTR8-SVneo細(xì)胞;(2)HTR8-SVneo細(xì)胞+poly(I:C);(3)HTR8-SVneo細(xì)胞+si RNA;(4)HTR8-SVneo細(xì)胞+si RNA STAT3;(5)HTR8-SVneo細(xì)胞+si RNA STAT3+poly(I:C);3 Western法測量(1)(3)(4)組STAT3及p-STAT3,了解是否轉(zhuǎn)染成功,且轉(zhuǎn)染本身對實(shí)驗(yàn)有無影響;測量(1)(2)組的STAT3及p-STAT3,了解Poly(I:C)在滋養(yǎng)細(xì)胞HTR8-SVneo中是否影響了STAT3的表達(dá);4 RT-PCR法測量(1)(2)(4)(5)組的s Flt-1,VEGF mRNA表達(dá)變化,了解STAT3在滋養(yǎng)細(xì)胞HTR8-SVneo中對VEGF及s Flt-1的影響。結(jié)果:1 SFlt-1 mRNA表達(dá)水平在Poly(I:C)(10ug/m1)刺激2h,4h,12h與對照組相比明顯升高(t1=5.59,P1=0.005;t2=3.75,P2=0.020;t3=2.979,P3=0.041);VEGF mRNA表達(dá)在刺激2h,4h,12h顯著受到抑制(t1=3.02,P1=0.039;t2=3.004,P2=0.040;t3=2.979,P3=0.410);從時(shí)效性來說,Poly(I:C)對s Flt-1、VEGF mRNA表達(dá)均在2h和4h效應(yīng)最明顯。2 HTR8-SVneo細(xì)胞在轉(zhuǎn)染STAT3 si RNA 48h后,Western blot檢測(1)(3)(4)組STAT3,p-STAT3,結(jié)果表明相對于(1)組及(3)組而言,轉(zhuǎn)染STAT3si RNA后HTR8-SVneo細(xì)胞中STAT3,p-STAT3蛋白明顯降低,提示轉(zhuǎn)染成功。3 Western blot檢測(1)(2)組STAT3,p-STAT3,結(jié)果表明(2)組與(1)組比較,STAT3表達(dá)無明顯變化,但p-STAT3明顯降低,有統(tǒng)計(jì)學(xué)意義;4(4)組與(1)組比較,s Flt-1mRNA顯著升高(t=3.66,P=0.021),VEGF mRNA顯著降低(t=3.03,P=0.038),提示STAT3在滋養(yǎng)細(xì)胞HTR8-SVneo中對VEGF、s Flt-1有影響;5 Poly(I:C)(10ug/ml)誘導(dǎo)后,(5)組表達(dá)s Flt-1mRNA較(2)組升高,有統(tǒng)計(jì)學(xué)意義(t=2.80,P=0.048),VEGFmRNA較(2)組降低,有統(tǒng)計(jì)學(xué)意義(t1=2.79,P=0.049),提示在Poly(I:C)可通過STAT3通路影響VEGF及s Flt-1。結(jié)論:1抑制STAT3通路可降低Poly(I:C)介導(dǎo)的滋養(yǎng)細(xì)胞VEGF表達(dá);2抑制STAT3通路可增強(qiáng)Poly(I:C)介導(dǎo)的滋養(yǎng)細(xì)胞s Flt-1表達(dá);3 STAT3信號通路改變可能影響滋養(yǎng)細(xì)胞的功能。
[Abstract]:Objective: abnormal expression of trophoblast may affect placental function and cause pregnancy related diseases. This study was to detect the changes of VEGF, s Flt-1 expression after the inhibition of STAT3 pathway in human chorionic trophoblast HTR8-SVneo cells under the induction of Poly (I:C), and to understand the effect of STAT3 pathway on the expression of nourishing cell VEGF and s Flt-1. Method: 1 Poly TR8-SVneo cells, cells that stimulate 0h, 2h, 4h, 6h, 12h, 24, RT-PCR detect s Flt-1, VEGF mRNA expression changes; 2 experiments are divided into five groups: (2) 3); (5) 4 cells (1) (1) (1) (3) (1) 4) group STAT3 and p-STAT3, to understand whether the transfection was successful, and whether the transfection was affected by the transfection itself; to measure the STAT3 and p-STAT3 of group (1) (2), to understand whether Poly (I:C) affects the expression of STAT3 in the trophoblast HTR8-SVneo; 4 RT-PCR method (1) (1) (2) (4) (5) s Flt-1. The effect of GF and s Flt-1. Results: 1 SFlt-1 mRNA expression level in Poly (10ug/m1) stimulates 2h, 4h, 12h is significantly higher than the control group. Poly (I:C) expressed s Flt-1 and VEGF mRNA in 2H and 4H effect most obviously.2 HTR8-SVneo cells after transfection STAT3 Si. Lot (1) (2) group STAT3, p-STAT3, the results showed that (2) compared with (1) group, the expression of STAT3 had no obvious change, but p-STAT3 significantly decreased, and there was a significant statistical significance. The 4 (4) group compared with (1) group, s Flt-1mRNA significantly increased (t=3.66, P=0.021), VEGF mRNA significantly decreased (t=3.03,). After the induction of 5 Poly (I:C) (10ug/ml), the expression of s Flt-1mRNA in group 5 was higher than that in group (2), and was statistically significant (t=2.80, P=0.048), VEGFmRNA was lower than that in group (2), and was statistically significant (t1=2.79, P=0.049), suggesting that Poly (I:C) could reduce the expression of nourishing cells mediated by the 1 inhibition pathway; 2 inhibition of STAT3 pathway can enhance Poly (I:C) - mediated s Flt-1 expression in trophoblastic cells, and 3 STAT3 signaling pathway may affect the function of trophoblast cells.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R714.2

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