CAJ | 학술논문

背景:近年的研究表明,肿瘤坏死因子α对不同组织成纤维细胞的作用具有组织特异性及浓度依赖性.目的:观察肿瘤坏死因子α及其信号传导途径中特异性激酶抑制剂对小鼠胚胎成纤维细胞成熟化所起的作用.方法:体外培养小鼠胚胎成纤维细胞,将细胞分为3组:第1组用含体积分数2%血清的DMEM高糖培养基培养作为空白对照组;第2组用含100 μg/L肿瘤坏死因子α的培养基培养;第3组先加入质量浓度为50 μg/L的Anti-TNFRSF1B作用1 h后,倒出培养基再加入含有肿瘤坏死因子α的培养基继续培养.采用RT-PCR法测定各组Ⅰ型胶原蛋白和基质金属蛋白酶3 mRNA表达、Western Blot法测定各组Ⅰ型胶原蛋白和基质金属蛋白酶3蛋白表达.结果与结论:小鼠胚胎成纤维细胞在一定质量浓度肿瘤坏死因子α作用下,其信号传导途径特异性激酶发生磷酸化或蛋白被激活,信号通路被激活,促进基质金属蛋白酶3的活化,明显降低Ⅰ型胶原的表达.加入其信号传导途径的抑制剂Anti-TNFRSF1B后,肿瘤坏死因子α的效应得到了一定的抑制,但并未完全消除,这更进一步证明肿瘤坏死因子α对小鼠胚胎成纤维细胞活化的作用.
배경:근년적연구표명,종류배사인자α대불동조직성섬유세포적작용구유조직특이성급농도의뢰성.목적:관찰종류배사인자α급기신호전도도경중특이성격매억제제대소서배태성섬유세포성숙화소기적작용.방법:체외배양소서배태성섬유세포,장세포분위3조:제1조용함체적분수2%혈청적DMEM고당배양기배양작위공백대조조;제2조용함100 μg/L종류배사인자α적배양기배양;제3조선가입질량농도위50 μg/L적Anti-TNFRSF1B작용1 h후,도출배양기재가입함유종류배사인자α적배양기계속배양.채용RT-PCR법측정각조Ⅰ형효원단백화기질금속단백매3 mRNA표체、Western Blot법측정각조Ⅰ형효원단백화기질금속단백매3단백표체.결과여결론:소서배태성섬유세포재일정질량농도종류배사인자α작용하,기신호전도도경특이성격매발생린산화혹단백피격활,신호통로피격활,촉진기질금속단백매3적활화,명현강저Ⅰ형효원적표체.가입기신호전도도경적억제제Anti-TNFRSF1B후,종류배사인자α적효응득도료일정적억제,단병미완전소제,저경진일보증명종류배사인자α대소서배태성섬유세포활화적작용.
BACKGROUND:In recent years,studies have shown the effects of tumor necrosis factor-a(TNF-а)on fibroblasts at different organizations in a tissue-specific and dose-dependent manner.OBJECTIVE:TO investigate the biologic effects of TNF-а and its signal transduction pathway-specific kinase inhibitors on mouse embryonic fibroblasts(NIH3T3).METHODS:NIH3T3 cells were cultured in vitro,and then the cells were assigned into 3 groups.Cells in the control group were cultured in DMEM high-glucose medium with 2%serum;those in the TNF-а group were cultured in 100 μg/L TNF-аmedium;those in the TNF-а+Anti-TNFRSF 1 B group were firstly cultured in medium with 50 μg/L Anti-TNFRSF 1 B for 1 hour,and then placed in the medium with 100 μg/L TNF-а.RT-PCR and Western blot methods were used to evaluate mRNA and protein expressions of type Ⅰ collagen and matrix metalloproteinase 3(MMP3)in each group.RESULTS AND CONCLUSION:In this experiment,NlH3T3 cells cultured with a certain concentration of TNF-а,the specificity kinase signal of transduction pathway presented with phosphorylation or protein activation,and the signal pathway was activated,which promoted MMP3 activation,and significantly reduced the expression of type Ⅰ collagen.The effect of TNF-а was certainly inhibited,but not completely eliminated after adding its signal transduction pathway inhibitor Anti-TNFRSF1 B.This further proves the role of TNF-а on NIH3T3 activation.