摘要
目的探讨肿瘤坏死因子-α(TNFα-)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-TNFRSF1B组(C组)。细胞制成2×108/L的细胞悬液后,接种于25 cm2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12~16 h。然后A组换用含体积分数0.02胎牛血清的DMEM高糖培养基继续培养;B组换用含100μg/LTNF-α的培养基培养;C组先加入浓度为50μg/L的Anti-TNFRSF1B作用1 h后,倒出培养基后再加入含100μg/L TNFα-的培养基继续培养。然后采用RT-PCR方法测定各组Ⅰ型胶原和基质金属蛋白酶3(MMP3)mRNA的表达,Western Blot方法测定各组Ⅰ型胶原蛋白和MMP3蛋白的表达。结果 B、C组MMP3 mRNA及蛋白的表达较A组升高,差异均有显著意义(t=-13.413~5.076,P<0.05);B组较C组升高,差异也具有显著意义(t=4.441~5.076,P<0.01);B、C组Ⅰ型胶原的表达较A组降低,差异也均有显著性(t=-4.950~5.808,P<0.05),B组较C组降低,差异也均有显著性(t=-4.950^-3.823,P<0.05)。结论 TNFα-可促进NIH3T3细胞活化。
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.
出处
《齐鲁医学杂志》
2010年第3期235-237,共3页
Medical Journal of Qilu
