Experimental Study Of Biologicd Effects Of Tumour Necrosis Factor-α On NIH3T3 Cells

Objective Investigate the biologic effects of tumour necrosis factor-α(TNF-α) and its signal transduction pathway-specific kinase inhibitors on NIH3T3 cells.Methods Culture NIH3T3 cells invitro, cells were divided into three groups.: group A,control group; group B,TNF-αgroup; group C,TNF-α+Anti-TNFRSF1B group.The cells first were made into 2×105/ml cell suspension, then were planted into 25 cm2 culture flask, when confluenting, the cells were maintained in DMEM high-glucose medium without serum for 12-16h. The control group was cultured in DMEM high-glucose medium with 2% serum; the TNF-agroup was cultured in 100μg/L TNF-αmedium; the TNF-α+Anti-TNFRSF1B group firstly were cultured in medium with 50μg/L Anti-TNFRSF1B for 1h, and then were cultured in medium with 100μg/L TNF-α.And then RT-PCR and Western blot methods were used to evaluated the expression of typeⅠcollagen and matrix metalloproteinase 3 (MMP3) mRNA and protein in each group.Results The results showed that either mRNA or protein expression of MMP3 in group B and group C compared with the group A were significantly different(t=-13.413-5.076,P＜0.05),group B compared with group C was also significant differences (t=4.441 -5.076, P＜0.01). The expression of both mRNA and protein of typeⅠcollagen in group B and group C compared with the group A were also significantly different (t=-4.950-5.808, P＜0.05), the statistic differences exist between group B and group C (t=-4.950--3.823,P＜0.05).Conclusion TNF-αpromoted the activation of NIH3T3 cells.