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目的探索白细胞介素-25(IL-25)通过nuocyte细胞对哮喘小鼠气道重塑的作用。方法 30只SPF级BALB/c雌性小鼠随机分为对照小鼠组、哮喘小鼠组、抗IL-25小鼠组,每组10只,鸡清卵蛋白(OVA)诱导哮喘模型。分离培养哮喘小鼠组nuocyte细胞,并分为对照细胞组、IL-25细胞组、抗IL-25细胞组。HE染色观察肺组织变化,流式细胞法计数肺泡灌洗液(BALF)中nuocyte细胞,ELISA法检测BALF及细胞上清液中IL-4、IL-13含量,免疫组化、Western blotting及RT-PCR法检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ胶原蛋白(collagen-Ⅰ)在肺组织中的表达。结果与对照小鼠组相比,哮喘小鼠组BALF中IL-4、IL-13表达升高,nuocyte细胞增多,α-SMA和collagen-Ⅰ表达升高(P<0.05),而抗IL-25小鼠组表达无明显变化(P>0.05);与对照细胞组相比,IL-25细胞组上清液IL-4、IL-13表达增高(P<0.05),抗IL-25细胞组上清液IL-4、IL-13表达无明显变化(P>0.05)。结论IL-25可能通过nuocyte细胞促使TH2型细胞因子分泌,导致哮喘小鼠气道重塑。
Objective To explore the effect of interleukin-25 (IL-25) on airway remodeling in asthmatic mice via nuocyte. Methods Thirty SPF BALB / c female mice were randomly divided into control group, asthma group and anti-IL-25 group, with 10 mice in each group. Asthma models were induced by chicken egg white (OVA). The nuocyte cells in asthmatic mice were isolated and cultured, and divided into control group, IL-25 group and anti-IL-25 group. The changes of lung tissue were observed by HE staining. The number of nuocyte in bronchoalveolar lavage fluid (BALF) was counted by flow cytometry. The levels of IL-4 and IL-13 in BALF and supernatant were measured by ELISA. Immunohistochemistry, Western blotting and RT -PCR method was used to detect the expression of α-smooth muscle actin (α-SMA) and collagen-Ⅰ in lung tissue. Results Compared with the control group, the expression of IL-4 and IL-13 in BALF of asthmatic mice increased, the number of nuocyte increased, the expression of α-SMA and collagen-Ⅰ increased (P <0.05) (P> 0.05). Compared with the control group, the expression of IL-4 and IL-13 in the supernatant of IL-25 group increased (P <0.05) The supernatant IL-4, IL-13 expression had no significant change (P> 0.05). Conclusion IL-25 may promote the secretion of TH2 cytokines by nuocyte cells, leading to airway remodeling in asthmatic mice.