STAT3信号通路对滋养细胞VEGF、sFlt-1表达的影响

发布时间：2018-05-23 07:57

本文选题：STAT3 + VEGF　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:滋养细胞表达异常可能影响胎盘功能,引发妊娠相关疾病。本研究通过检测在Poly(I:C)诱导下抑制人绒毛滋养层细胞HTR8-SVneo细胞中STAT3通路后VEGF、s Flt-1表达的变化,了解STAT3通路对滋养细胞VEGF、s Flt-1表达的影响。方法:1 Poly(I:C)刺激HTR8-SVneo细胞,收集刺激0h、2h、4h、6h、12h、24后的细胞,RT-PCR检测s Flt-1,VEGF mRNA表达变化;2实验分为五组:(1)HTR8-SVneo细胞;(2)HTR8-SVneo细胞+poly(I:C);(3)HTR8-SVneo细胞+si RNA;(4)HTR8-SVneo细胞+si RNA STAT3;(5)HTR8-SVneo细胞+si RNA STAT3+poly(I:C);3 Western法测量(1)(3)(4)组STAT3及p-STAT3,了解是否转染成功,且转染本身对实验有无影响;测量(1)(2)组的STAT3及p-STAT3,了解Poly(I:C)在滋养细胞HTR8-SVneo中是否影响了STAT3的表达;4 RT-PCR法测量(1)(2)(4)(5)组的s Flt-1,VEGF mRNA表达变化,了解STAT3在滋养细胞HTR8-SVneo中对VEGF及s Flt-1的影响。结果:1 SFlt-1 mRNA表达水平在Poly(I:C)(10ug/m1)刺激2h,4h,12h与对照组相比明显升高(t1=5.59,P1=0.005;t2=3.75,P2=0.020;t3=2.979,P3=0.041);VEGF mRNA表达在刺激2h,4h,12h显著受到抑制(t1=3.02,P1=0.039;t2=3.004,P2=0.040;t3=2.979,P3=0.410);从时效性来说,Poly(I:C)对s Flt-1、VEGF mRNA表达均在2h和4h效应最明显。2 HTR8-SVneo细胞在转染STAT3 si RNA 48h后,Western blot检测(1)(3)(4)组STAT3,p-STAT3,结果表明相对于(1)组及(3)组而言,转染STAT3si RNA后HTR8-SVneo细胞中STAT3,p-STAT3蛋白明显降低,提示转染成功。3 Western blot检测(1)(2)组STAT3,p-STAT3,结果表明(2)组与(1)组比较,STAT3表达无明显变化,但p-STAT3明显降低,有统计学意义;4(4)组与(1)组比较,s Flt-1mRNA显著升高(t=3.66,P=0.021),VEGF mRNA显著降低(t=3.03,P=0.038),提示STAT3在滋养细胞HTR8-SVneo中对VEGF、s Flt-1有影响;5 Poly(I:C)(10ug/ml)诱导后,(5)组表达s Flt-1mRNA较(2)组升高,有统计学意义(t=2.80,P=0.048),VEGFmRNA较(2)组降低,有统计学意义(t1=2.79,P=0.049),提示在Poly(I:C)可通过STAT3通路影响VEGF及s Flt-1。结论:1抑制STAT3通路可降低Poly(I:C)介导的滋养细胞VEGF表达;2抑制STAT3通路可增强Poly(I:C)介导的滋养细胞s Flt-1表达;3 STAT3信号通路改变可能影响滋养细胞的功能。
[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.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.2

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