黄芩素上调miR-625-5p表达 干预支气管哮喘小鼠的机制研究

The mechanism of baicalein up-regulating miR-625-5p expression in bronchial asthma mice

目的:探讨黄芩素调控miR-625-5p表达对支气管哮喘小鼠气道平滑肌细胞(ASMCs)增殖、迁移及炎性因子的影响。方法:培养小鼠原代ASMCs细胞,作为对照组,重组小鼠血小板衍生生长因子(PDGF)诱导ASMCs增殖,作为PDGF组;分别采用10、20、40μmol/L的黄芩素处理PDGF诱导ASMCs细胞。将miR-NC、miR-625-5p转染至PDGF诱导的ASMCs细胞,作为PDGF+miR-NC、PDGF+miR-625-5p组;将anti-miR-NC、anti-miR-625-5p转染至PDGF诱导ASMCs细胞,以40μmol/L的黄芩素处理,作为PDGF+anti-miR-NC+40μmol/L组、PDGF+anti-miR-625-5p+40μmol/L组。CCK-8法检测细胞增殖;Transwell实验检测细胞迁移;酶联免疫吸附实验检测细胞IL-4、IL-6、IL-8水平;实时荧光定量PCR检测细胞miR-625-5p表达水平。结果:与对照组比较,PDGF组细胞增殖率、迁移率以及IL-4、IL-6、IL-8水平显著升高(均P<0.05);与PDGF组比较,黄芩素低、中、高剂量组细胞增殖率、迁移率以及IL-4、IL-6、IL-8水平显著降低(均P<0.05)。与对照组比较,PDGF组细胞miR-625-5p表达显著降低(P<0.05);与PDGF组比较,黄芩素低、中、高剂量组细胞miR-625-5p表达显著升高(均P<0.05)。与PDGF+miR-NC组比较,PDGF+miR-625-5p组细胞增殖率、迁移率以及IL-4、IL-6、IL-8水平显著降低(均P<0.05)。与PDGF+anti-miR-NC+40μmol/L组比较,PDGF+anti-miR-625-5p+40μmol/L组细胞miR-625-5p表达水平显著降低,细胞增殖率、迁移率以及IL-4、IL-6、IL-8水平显著升高(均P<0.05)。结论:黄芩素通过上调miR-625-5p表达抑制支气管哮喘小鼠的细胞增殖率、迁移率以及炎性因子水平。

Objective:To investigate the effect of baicalein on the proliferation,migration and inflammatory factors of airway smooth muscle cells(ASMCs)in bronchial asthma mice by regulating the expression of miR-625-5p.Methods:Primary mouse ASMCs cells were cultured,as control group,recombinant mouse platelet-derived growth factor(PDGF)induced the proliferation of ASMCs,as PDGF group;10,20,40μmol/L of baicalein treated PDGF to induce ASMCs cells.miR-NC and miR-625-5p were transfected into PDGF-induced ASMCs cells as PDGF+miR-NC,PDGF+miR-625-5p group;anti-miR-NC and anti-miR-625-5p were transfected ASMCs cells were induced by PDGF,and then treated with 40μmol/L of baicalein,as PDGF+anti-miR-NC+40μmol/L group,PDGF+anti-miR-625-5p+40μmol/L group.Cell proliferation was detected by CCK-8 method;Cell migration was detected by Transwell assay;IL-4,IL-6 and IL-8 levels were detected by enzyme-linked immunosorbent assay;Real-time quantitative PCR to detect the expression level of miR-625-5p in cells.Results:Compared with the control group,the cell proliferation rate,migration rate and the levels of IL-4,IL-6 and IL-8 in the PDGF group were significantly increased(all P<0.05).Compared with PDGF group,the proliferation rate,migration rate and IL-4,IL-6 and IL-8 levels of baicalin cells at were significantly decreased(all P<0.05).Compared with the control group,the expression of miR-625-5p in the PDGF group was significantly decreased(P<0.05);compared with the PDGF group,the expression of miR-625-5p in was significantly increased(all P<0.05).Compared with the PDGF+miR-NC group,the cell proliferation rate,migration rate and the levels of IL-4,IL-6 and IL-8 in the PDGF+miR-625-5p group were significantly decreased(all P<0.05).Compared with PDGF+anti-miR-NC+40μmol/L group,the expression level of miR-625-5p in PDGF+anti-miR-625-5p+40μmol/L group was significantly decreased,and the cell proliferation rate,migration rate and IL-4,IL-6,IL-8 levels were significantly increased(all P<0.05).Conclusion:Baicalein inhibited the proliferation rate,migration rate and inflammatory factors of bronchial asthma in mice by up-regulating the expression of miR-625-5p.