布地奈德通过下调miR-620抑制PDGF诱导的小鼠气道平滑肌细胞增殖和炎性因子表达

Budesonide inhibiting PDGF-induced mouse airway smooth muscle cell proliferation and expression of inflammatory factor by down-regulating miR-620

目的探讨布地奈德(Bud)对血小板衍生生长因子(PDGF)诱导的小鼠气道平滑肌细胞(ASMC)增殖和炎性因子表达的影响及可能机制。方法体外培养小鼠ASMC,不同剂量(200、400、600μg/mL)的Bud作用于PDGF诱导的ASMC、PDGF诱导转染miR-620抑制剂的ASMC或600μg/mL的Bud作用于PDGF诱导的转染miR-620模拟物的ASMC,CCK-8法检测细胞增殖、Western Blot检测细胞中CyclinD1和P21蛋白表达,试剂盒检测细胞培养上清液中IL-4、IL-5和IL-13水平,RT-qPCR检测细胞中miR-620表达。结果不同浓度(200、400、600μg/mL)的Bud降低了PDGF诱导的ASMC细胞OD值及CyclinD1蛋白和miR-620表达(P<0.05),而促进了P21蛋白表达(P<0.05),降低了细胞培养上清液中IL-4、IL-5和IL-13水平(P<0.05)。沉默miR-620降低了PDGF诱导的ASMC细胞OD值及CyclinD1蛋白表达(P<0.05),促进了P21蛋白表达(P<0.05),降低了细胞培养上清液中IL-4、IL-5和IL-13水平(P<0.05)。过表达miR-620逆转了600μg/mL的Bud对PDGF诱导的ASMC细胞OD值、CyclinD1和P21的蛋白及IL-4、IL-5和IL-13表达的影响(P<0.05)。结论Bud可能通过下调miR-620表达,抑制PDGF诱导的小鼠气道平滑肌细胞增殖和炎性因子表达。

Objective To investigate the effect of budesonide(Bud)on the proliferation and the expression of inflammatory factors in mouse airway smooth muscle cells(ASMC)induced by platelet-derived growth factor(PDGF)and its possible mechanism.Methods Mouse ASMCs were cultured in vitro.Different doses(200,400,600μg/mL)of Bud were given to PDGF-induced ASMC,PDGF-induced transfection of miR-620 inhibitor ASMC or 600 Bud acted on PDGF-induced transfection of miR-620 simulated ASMC.CCK-8 method was used to detect cell proliferation,western blot was used to detect the protein expression of CyclinD1 and P21 in cells,kits were used to detect the levels of IL-4,IL-5 and IL-13 in cell culture supernatant,and RT-qPCR was used to detect the expression of miR-620 in cells.Results Bud at different concentrations(200,400,600μg/mL)reduced the OD value of PDGF-induced ASMC cells and the expression of CyclinD1 protein and miR-620(P<0.05),promoted the expression of P21 protein(P<0.05),and reduced the levels of IL-4,IL-5 and IL-13 in the culture supernatant(P<0.05).Silencing miR-620 reduced the OD value of PDGF-induced ASMC cells and the expression of CyclinD1 protein(P<0.05),promoted the expression of P21 protein(P<0.05),and reduced the levels of IL-4,IL-5 and IL-13 in the culture supernatant(P<0.05).Over-expressing miR-620 reversed the effect of 600μg/mL Bud on PDGF-induced ASMC cell OD value,the protein expression of CyclinD1 and P21,and the levels of IL-4,IL-5 and IL-13(P<0.05).Conclusion Bud may inhibit PDGF-induced airway smooth muscle cell proliferation and the expression of inflammatory factor by down-regulating miR-620.