辛前甘桔汤对慢性鼻窦炎小鼠lncRNA SNHG16、EGFR及MUC5AC mRNA表达的影响

Effects of Xinqian Ganjie Decoction on lncRNA SNHG16,EGFR and MUC5AC mRNA expression in chronic rhinosinusitis mice

目的研究辛前甘桔汤对慢性鼻窦炎(CRS)小鼠长链非编码RNA小核仁RNA宿主基因16(lnc RNA SNHG16)、表皮生长因子受体(EGFR)及黏蛋白5AC(MUC5AC)m RNA表达的影响。方法36只C57BL/6小鼠随机分为正常组、模型组、羧甲司坦组及辛前甘桔汤低、中、高剂量组,每组6只。采用金黄色葡萄球菌鼻腔灌注建立小鼠CRS模型。正常组与模型组小鼠灌胃质量分数为0.9%的氯化钠溶液(0.6 g·kg^(-1)·d^(-1)),辛前甘桔汤低、中、高剂量组分别灌胃辛前甘桔汤水煎液(0.3、0.6、1.2 g·kg^(-1)·d^(-1)),羧甲司坦组灌胃羧甲司坦口服液(0.3 m L·kg^(-1)·d^(-1)),连续干预14 d。采用苏木精-伊红(HE)及阿尔新蓝-过碘酸雪夫(AB-PAS)染色法观察小鼠鼻黏膜结构,免疫组织化学法观察小鼠鼻黏膜EGFR、MUC5AC表达并测量光密度值;酶联免疫吸附试验(ELISA)检测样本中白介素-6(IL-6)、白介素-8(IL-8)、基质金属蛋白酶-9(MMP-9)水平。实时荧光定量逆转录聚合酶链式反应(RT-q PCR)检测样本中lnc RNA SNHG16、EGFR、MUC5AC m RNA的表达。结果与正常组比较,模型组鼻黏膜上皮结构紊乱,IL-6、IL-8、MMP-9含量明显升高(P<0.05),lnc RNA SNHG16、EGFR、MUC5AC m RNA表达明显升高(P<0.05)。与模型组比较,辛前甘桔汤各剂量组IL-6水平显著降低(P<0.05),辛前甘桔汤中、高剂量组MMP-9、IL-8水平降低(P<0.05),辛前甘桔汤各剂量组lnc RNA SNHG16、EGFR、MUC5AC m RNA表达显著降低(P<0.05)。与羧甲司坦组比较,辛前甘桔汤高剂量组lnc RNA SNHG16、EGFR m RNA表达降低(P<0.05)。结论辛前甘桔汤能降低lnc RNA SNHG16、EGFR、MUC5AC m RNA表达,降低EGFR、MUC5AC蛋白表达,减少IL-6、IL-8、MMP-9表达,进而改善鼻黏膜病理改变。

Objective To Study the effects of Xinqian Ganjie Decoction(XQGJD)on long strand non coding RNA small nucleolar RNA host gene 16(lncRNA SNHG16),epidermal growth factor receptor(EGFR)and mucin 5AC(MUC5AC)mRNA expression in mice with chronic rhinosinusitis(CRS).Methods Thirty-six C57BL/6 mice were randomly divided into six groups:normal group,model group,low-dose XQGJD group,medium-dose XQGJD group,high-dose XQGJD group and Carbocisteine group,with 6 mice in each group.The CRS model was established by intranasal infusion of Staphylococcus aureus.Mice in the normal group and model group were given 0.9%NaCl solution by gavage(0.6 g·kg^(-1)·d^(-1)),mice in the XQGJD groups were treated with XQGJD by gavage(0.3,0.6,and 1.2 g·kg^(-1)·d^(-1) respectively),mice in the Carbocisteine group was treated with Carbocisteine by gavage(0.3 mL·kg^(-1)·d^(-1)).The treatment lasted for 14 days.The HE and AB-PAS staining were used to evaluate the structure of mouse nasal mucosa.The immunohistochemical staining was used to observe the expressions of EGFR and MUC5AC,and optical density IOD values were measured.The contents of interleukin-6(IL-6),interleukin-8(IL-8),and human matrix metalloproteinase-9(MMP-9)were determined by ELISA.The expressions of lncRNA SNHG16,EGFR,MUC5AC mRNA were detected by RT-qPCR.Results Compared with the normal group,the nasal mucosa epithelium in the model group was in disorder,with increased IL-6,IL-8,MMP-9 secretion(P<0.05),increased lncRNA SNHG16,EGFR,MUC5AC mRNA expressions(P<0.05).Compared with the model group,the level of IL-6 in each dose group of XQGJD was significantly lower(P<0.05),the levels of MMP-9 and IL-8 in the medium and high dose groups of XQGJD were lower(P<0.05),and the expressions of lncRNA SNHG16,EGFR,MUC5AC mRNA in each dose group of XQGJD were significantly lower(P<0.05).Compared with Carbocisteine group,the expressions of lncRNA SNHG16 mRNA and EGFR mRNA in the high-dose XQGJD group were decreased(P<0.05).Conclusion Xinqian Ganjie Decoction can reduce expressions of lncRNA SNHG16,EGFR and MUC5AC mRNA and EGFR and MUC5AC proteins,lower IL-6,IL-8 and MMP-9 secretion,and improve the pathological changes of nasal mucosa in chronic sinusitis model mice.