ADAM33对人气道平滑肌细胞增殖、凋亡的作用及IL-4对其表达的影响

发布时间：2018-04-06 06:22

本文选题：解整合素-金属蛋白酶　切入点：哮喘　出处：《上海交通大学学报(医学版)》2014年07期

【摘要】：目的探讨解整合素-金属蛋白酶33(ADAM33)对人气道平滑肌细胞(HASMCs)增殖和凋亡的作用,以及白介素-4(IL-4)对ADAM33表达的影响。方法分别以1、10、50和100 ng/mL浓度的IL-4刺激HASMCs,以未刺激细胞组作为对照,分别采用Real-Time PCR和Western blotting法检测ADAM33 mRNA和蛋白的表达。设计并合成ADAM33-siRNA,瞬时转染HASMCs,Real-Time PCR法和Western blotting法筛选出抑制率最高的siRNA,以抑制率最高的siRNA瞬时转染HASMCs;MTS和流式细胞术分别检测细胞的增殖和凋亡情况。结果当不同浓度的IL-4刺激HASMCs时,ADAM33 mRNA和蛋白的表达均呈浓度依赖性。与对照组比较,IL-4浓度为50 ng/mL组和100 ng/mL组mRNA和蛋白的相对表达量分别增加了5.94、7.6倍和3.72、4.57倍,差异均有统计学意义(P0.05);但1 ng/mL组和10 ng/mL组无明显升高,差异均无统计学意义(P0.05)。在3条干扰RNA中,ADAM33-siRNA-575的抑制效率最高,在mRNA水平的抑制率为81.08%。在24、48和72 h,干扰组的细胞增殖明显低于阴性对照组(P0.05),增殖抑制率分别为15.38%、32.83%和22.78%;干扰组的细胞凋亡率为(25.88±3.37)%,明显高于阴性对照组的(8.95±0.51)%(P0.05)。结论 ADAM33可促进HASMCs增殖,而细胞因子IL-4可促进其表达,两者可能在气道重塑中起着重要的作用。
[Abstract]:Objective to investigate the effect of integrin-metalloproteinase-33 (ADAM33) on proliferation and apoptosis of human airway smooth muscle cells (HASMCs) and the effect of interleukin-4 (IL-4) on the expression of ADAM33.Methods the expression of ADAM33 mRNA and protein were detected by Real-Time PCR and Western blotting in the unstimulated cells group and the unstimulated cells group, respectively. The HASMCs were stimulated with IL-4 at the concentrations of 1 ng/mL and 100 ng/mL, and the expression of ADAM33 mRNA and protein were detected by Real-Time PCR and Western blotting respectively.ADAM33-siRNAs were designed and synthesized. The siRNAs with the highest inhibition rate were screened by transient transfection of HASMCsSU Real-Time PCR and Western blotting. The proliferation and apoptosis of the cells were detected by flow cytometry and transient transfection of siRNA with the highest inhibition rate.Results the expression of ADAM33 mRNA and protein was in a concentration-dependent manner when HASMCs was stimulated with different concentrations of IL-4.Compared with the control group, the relative expression of mRNA and protein in 50 ng/mL group and 100 ng/mL group increased 5.947.6-fold and 3.72U 4.57 times, respectively, and the difference was statistically significant (P 0.05), but 1 ng/mL group and 10 ng/mL group had no significant increase, the difference was not statistically significant (P 0.05).Among the three interfering RNA, ADAM33-siRNA-575 had the highest inhibitory efficiency, and the inhibition rate at mRNA level was 81.08%.At 24: 48 and 72 h, the cell proliferation in the interference group was significantly lower than that in the negative control group (P 0.05), the inhibition rate of proliferation was 15.38% and 32.83%, and the cell apoptosis rate in the interference group was 25.88 卤3.37%, which was significantly higher than that in the negative control group (P 0.05).Conclusion ADAM33 can promote the proliferation of HASMCs and cytokine IL-4 can promote its expression. Both of them may play an important role in airway remodeling.
【作者单位】：重庆市涪陵中心医院呼吸内科;
【基金】：重庆市卫生局基金(07-2-222)~~
【分类号】：R562.25

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