微小 RNA-21在小鼠肺成纤维细胞炎症反应中的表达变化及意义

摘要

Objective To investigate the expression changes of microRNAs-21 ( miR-21 ) and its downstream target genes ( a disintegrin and metalloprotease with thrombospond in type 1 motifs, ADAMTS-1) in inflammation reaction of lung fibroblasts cells of mice and the clinical significance .Methods The fibroblast cell line NIH3T3 of mic+9e were cultured in vitro and then were divided into the control group and the TGF-β1 group.The TGF-β1 group was added with 5 μg/L TGF-β1 for stimulating 24, 48, 72, 96 and 120 h.The control group was not treated .The supernatant of the two groups was collected at different time points .Using the enzyme linked immunosorbent assay (ELISA) to detect the levels of tumor necrosis factor-α(TNF-α), inflammatory cytokine interleukin-1β(IL-1β) and interleukin-6 (IL-6).The cells were col-lected at different time points .We used the real-time fluorescence quantitative polymerase chain reaction ( PCR) to detect the expression of miR-21 and ADAMTS-1.Results The expression levels of TNF-a , IL-1β, IL-6 and miR-21 in the TGF-β1 groups at 24, 48, 72, 96 and 120 h were higher than those of the control group;the expression level of ADAMTS-1 was lower than that of the control group (all P<0.05 or P<0.01).Conclusions In the TGF-β1-induced inflammatory responses of lung fibroblast cells of mice , the miR-21 expression is up-regulated, while the ADAMTS-1 expression is down-regulated;miR-21 may be a new molecular target for the prevention and treatment of pulmonary fibrosis.%目的：观察小鼠肺成纤维细胞炎症反应中微小RNA-21（ miR-21）及其下游靶基因含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶（ADAMTS-1）的表达变化，并探讨其临床意义。方法体外培养小鼠NIH3T3成纤维细胞，分为对照组和TGF-β1组。 TGF-β1组加入终浓度为5μg／L的TGF-β1，分别作用24、48、72、96、120 h。对照组不加入TGF-β1。收集两组上清液，采用ELISA法测定TNF-α、IL-1β、IL-6水平，应用qRT-PCR技术检测miR-21及ADAMTS-1的表达。结果 TGF-β1组24、48、72、96、120 h时TNF-α、IL-1β、IL-6水平及miR-21表达均高于对照组，ADAMTS-1表达均低于对照组（P均＜0．05或0．01）。结论在TGF-β1诱导的大鼠肺成纤维细胞的炎症反应中，miR-21表达增高、ADAMTS-1表达降低；miR-21可成为预防和治疗肺纤维化新的分子靶点。