THE EFFECT OF TUMOUR NECROSIS FACTOR-α ON MATURITY OF NIH3T3 CELLS

Objective To study the effect of tumour necrosis factor-α(TNF-α) on the maturity of NIH3T3 cells. Methods NIH3T3 cells were cultured in vitro,and divided into three groups:group A(control group),group B(group TNF-α) and group C(group TNF-α+Anti-TNFRSF1B).After the cell suspension of 2×108/L had been made,it was then put into 25 cm2 culture flask.When the cells were confluented,they were cultured in serum-free DMEM high-glucose medium for 12-16 hours.The group A was then changed to be cultured in DMEM high-glucose medium with 2% serum;the group B was changed to be cultured in 100 μg/L TNF-α medium;and the group C was cultured in medium with 50 μg/L Anti-TNFRSF1B for one hour and then cultured in medium with 100 μg/L TNF-α.RT-PCR and Western Blot method were applied to detect the expressions of typeⅠcollagen and matrix metalloproteinase 3(MMP3) mRNA and protein in each group. Results The expressions of MMP3 mRNA and protein in groups B and C were significantly different from that in group A(t=-13.413-5.076,P0.05),the difference between groups B and C was also significant(t=4.441-5.076,P0.01).The expressions of both mRNA and protein of type Ⅰ collagen in groups B and C showed difference as compared with the group A(t=-4.950-5.808,P0.05),the statistic difference between groups B and C was also significant(t=-4.950——3.823,P0.05). Conclusion TNF-α can promote the activation of NIH3T3 cells.