吡美莫司对特异性皮炎合并哮喘小鼠气道重塑的影响研究

Effect of pimecrolimus on interleukin-33 level in mice with atopic dermatitis and asthma

目的观察吡美莫司(PYM)对特异性皮炎合并哮喘小鼠气道重塑的影响.方法健康雌性BALB/c小鼠21只,随机分成对照组、特异性皮炎组和吡美莫司治疗组.瑞氏-吉姆萨染色法计算支气管肺泡灌洗液(BALF)中白细胞总数及白细胞分类计数、苏木精-伊红(HE)染色观察肺组织和皮肤组织炎细胞浸润和组织结构变化、酶联免疫吸附法检测血清中白细胞介素(IL)-33、IL-5和IL-13表达水平.Western blot法观察P38丝裂原激活蛋白激酶(P38MAPK)和应激激活的蛋白激酶1的磷酸化水平.实时荧光定量聚合酶链式反应(PCR)及Western blot法检测Ⅰ型胶原蛋白(Col1)的mRNA和蛋白表达水平.结果与对照组小鼠相比,特异性皮炎组小鼠BALF中白细胞总数、嗜酸性粒细胞数目、血清IL-33、IL-13和IL-5表达水平均明显增高,差异有统计学意义(P<0.05),特异性皮炎组小鼠肺组织中P38MAPK及MSK1蛋白磷酸化增强(1.50±0.43比0.80±0.43、1.390±0.078比0.620±0.082),Col1mRNA及蛋白表达增加(1比3.2±0.59、1.40±0.12比0.16±0.13),差异有统计学意义(P<0.05).吡美莫司外部涂抹治疗后小鼠BALF中白细胞数目和血清IL-33、IL-5、IL-13水平下降、肺组织中P38MAPK及MSK1蛋白磷酸化减弱(0.10±0.04比0.85±0.05)、Col1mRNA及蛋白表达下降(0.61±0.22比0.48±0.08),差异有统计学意义(P<0.05),肺组织和皮肤组织水肿及炎性细胞浸润较前减轻.结论吡美莫司通过降低血清IL-33水平,抑制IL-33相关通路的活化,从而缓解小鼠皮肤和气道炎性反应及气道重塑,进而减少哮喘的发病.

Objective To explore the potential mechanism of pimecrolimus in airway remodeling of atopic dermatitis(AD)and asthma mice.Methods Female BALB/c mice were randomly divided into three groups:control group,atopic dermatitis group and pimecrolimus treatment group.The cell in bronchoalveolar lavage fluid(BALF)was calculated by Ray-Jimsa staining.The structural changes in lung tissue and skin tissue were observed by hematoxylin-eosin(HE)staining,and the level of interleukin(IL)-33,IL-5,IL-13 in serum was detected by enzyme-linked immunosorbent assay(ELISA).The expressions of type 1 collagen(Col1)in the airway of mice were detected by RT-PCR and Western blotting.And western blotting was also used to determine the activation of mitogen-activated protein kinase P38(P38MAPK)and mitogen-and stress-activated protein kinase 1(MSK1)in the lungs of mice.Results Compared with those in the control group,the total number of leukocytes in bronchoalveolar lavage fluid(BALF)and the expression levels of IL-33,IL-13 and IL-5 in serum of atopic dermatitis group were significantly higher,and the difference was statistically significant(P<0.05).In the atopic dermatitis group,the phosphorylation of P38MAPK and MSK1 protein was increased(1.50±0.43 vs.0.80±0.43,1.39±0.08 vs.0.62±0.08)and the expression of Col1 protein and protein(1 vs.3.20±0.59,1.40±0.12 vs.0.13±0.16)was increased,and the difference was statistically significant(P<0.05).After pimecrolimus treatment,the number of leukocytes in BALF,the levels of IL-33,IL-5 and IL-13 in serum(213.13±11.89,657.97±86.47,143.82±33.02),the phosphorylation of P38MAPK and MSK1 protein in lung tissue(0.10±0.04,0.85±0.05),and the expression of Col1 mRNA and Col1 protein were decreased(0.61±0.22,0.48±0.08),and the difference was statistically significant(P<0.05).The edema of lung and skin tissue and the infiltration of inflammatory cells were alleviated.Conclusions Pimecrolimus can alleviate the inflammation and airway remodeling in mice by inhibiting the activation of IL-33 related pathways,thereby reducing the incidence of asthma.