肿瘤坏死因子-α对NIH3T3细胞成熟化作用的实验研究

发布时间：2018-06-17 14:14

  本文选题：肿瘤坏死因子α + NIH3T3细胞　； 参考：《青岛大学》2010年硕士论文

【摘要】： 目的：探讨肿瘤坏死因子-α(TNF-α)及其信号传导途径中特异性激酶抑制剂对NIH3T3细胞成熟化所起的作用。 方法：体外培养NIH3T3成纤维细胞,将细胞分为空白对照组(A组)、TNF-α组(B组)及TNF-α+Anti-TNFRSFIB组(C组)。细胞制成2×105/mL的细胞悬液后,接种于25cm2培养瓶中,待细胞呈汇合状态时,换用无血清DMEM高糖培养基培养12-16h。然后A组换用含体积分数为0.02胎牛血清的DMEM高糖培养基继续培养；B组换用含100μg/L TNF-α的培养基培养；C组先加入浓度为50u g/L的Anti-TNFRSFIB作用于细胞1h后,倒出培养基后再加入含100μg/L TNF-α的培养基继续培养。然后采用RT-PCR法测定各组Ⅰ型胶原和基质金属蛋白酶3 (MMP3) mRNA的表达,Western Blot法测定各组Ⅰ型胶原蛋白和MMP3蛋白的表达。 结果：MMP3无论是mRNA还是蛋白的表达B组和C组与A组相比均有显著性差异(t=-13.413-5.076,P0.05)。其中B组与C组相比,差异也具有显著性(t=4.441-5.076,P0.01)；Ⅰ型胶原二者的表达B组和C组与A组相比也均有显著性差异(t=-4.950-5.808,P0.05),B组与C组民较,差异显著(t=-4.950--3.823,P0.05)。 结论：与对照组相比,经TNF-α干预后,NIH3T3成纤维细胞中Ⅰ型胶原蛋白表达明显减少,而MMP3的表达量则增多。TNF-α促进了NIH3T3的活化。
[Abstract]:Aim: to investigate the effect of specific kinase inhibitor on the maturation of NIH3T3 cells by tumor necrosis factor- 伪 (TNF- 伪) and its signal transduction pathway. Methods: NIH3T3 fibroblasts were cultured in vitro. The cells were divided into two groups: control group (group A) and TNF- 伪 Anti-TNFRSFIB group (group C). Cells were prepared into 2 脳 105 / mL cell suspensions and inoculated in 25cm2 culture flask. When the cells were confluent, the cells were cultured in serum-free high glucose medium for 12-16 hours. Then group A was treated with DMEM high sugar medium containing 0.02 fetal bovine serum by volume fraction. Group B was transferred to culture medium containing 100 渭 g / L TNF- 伪. Group C was treated with 50 u g / L Anti-TNFRSFIB for 1 h. Culture medium containing 100 渭 g / L TNF- 伪 was added. Then the mRNA expression of collagen 鈪，

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