目的 探讨KDM2A在肺纤维化进展中的表达变化,以期为寻求肺纤维化的发病机制提供具有潜力的新方向.方法 将8～10周的C57BL/6J雄性小鼠随机分配为正常组、14d模型组与21 d模型组.取模型组小鼠麻醉后,气道滴定博来霉素诱导建立肺纤维化模型.分别于造模后第14天、第21天取材14 d模型组与21 d模型组小鼠肺组织,最后取材正常组小鼠肺组织.通过HE染色,Masson染色来评估小鼠肺泡炎及纤维化程度.免疫组化分析3组肺组织α-SMA、FN、TGF-β1及KDM2A蛋白表达情况并进行免疫组化评分.Western Blot法检测α-SMA、FN、TGF-β1及KDM2A蛋白表达量.结果 与正常组比较,14 d模型组肺泡炎评分明显增加(P＜0.001);21 d模型组较14 d模型组比较,肺泡炎评分轻度增加,但差异无统计学意义(P＞0.05).与正常组比较,14 d模型组纤维化评分明显增加(P＜0.01);21 d模型组较14 d模型组比较,纤维化评分明显增加(P＜0.01).免疫组织化学染色结果显示:与正常组比较,14 d 模型组 α-SMA(P＜0.01)、FN(P＜0.01)、TGF-β1(P＜0.01)、KDM2A(P＜0.01)蛋白免疫组化评分明显升高;21 d 模型组较 14 d 模型组比较,α-SMA(P＜0.01)、FN(P＜0.05)、TGF-β1(P＜0.01)、KDM2A(P＜0.01)蛋白免疫组化评分升高.Western Blot 结果显示:与正常组比较,14 d 模型组 α-SMA(P＜0.01)、FN(P＜0.01)、TGF-β1(P＜0.01)、KDM2A(P＜0.01)蛋白表达量明显升高;21 d 模型组较 14 d 模型组相比,α-SMA(P＜0.01)、FN(P＜0.01)、TGF-β1(P＜0.01)、KDM2A(P＜0.01)蛋白表达量升高.结论 KDM2A在BLM诱导的肺纤维化小鼠肺组织中表达增强,其可能通过调控TGF-β1的表达来参与肺纤维化的形成.
Expression changes of KDM2A in bleomycin-induced pulmonary fibrosis in mice
Objective To investigate the expression changes of KDM2A in the progression of pulmonary fibrosis,thus providing a potential new direction for unveiling the pathogenesis of pulmonary fibrosis.Methods Male C57BL/6J mice from 8 weeks to 10 weeks of age were randomly assigned to 3 groups:normal group(n=11),14d model group(n=11)and 21 d model group(n=11).After anesthesia,intratracheal bleomycin(BLM)was used to induce pulmonary fibrosis in the model group.On the 14th and 21st day after modeling,mouse lung tissues in the 14d model group and 21 d model group were collected,respectively.Mouse lung tissues in the normal group were collected at the end of the experimental period.The hematoxylin and eosin(H&E)staining and Masson staining were used to evaluate the degree of alveolitis and fibrosis in mice,respectively.The protein expressions of alpha-smooth muscle actin(α-SMA),fibronectin(FN),transforming growth factor-beta1(TGF-β1)and KDM2A in lung tissues were analyzed by immunohistochemistry,and immunohistochemical scores were graded.The protein expression levels of α-SMA,FN,TGF-β1 and KDM2A were detected by Western blot as well.Results Compared with that of the normal group,the alveolitis score in the 14d model group was significantly higher(P＜0.01).It was higher in the 21 d model group than that of 14d model group,but the difference was not statistically significant(P＞0.05).Compared with that of the normal group,the fibrosis score of the 14d model group was significantly higher(P＜0.01).It was significantly higher in the 21d model group than that of 14d model group(P＜0.01).Immunohistochemical scores of α-SMA(P＜0.01),FN(P＜0.01),TGF-β1(P＜0.01)and KDM2A(P＜0.01)in 14d model group were significantly higher than those of normal group.Immunohistochemical scores of α-SMA(P＜0.01),FN(P＜0.05),TGF-β1(P＜0.01)and KDM2A(P＜0.01)in the 21 d model group were significantly higher than those of 14d model group.Western blot results showed that compared with those of the normal group,the protein expressions of α-SMA(P＜0.01),FN(P＜0.01),TGF-p1(P＜0.01)and KDM2A(P＜0.01)in the 14d model group were significantly upregulated.The protein expressions ofα-SMA(P＜0.01),FN(P＜0.01),TGF-β1(P＜0.01)and KDM2A(P＜0.01)were significantly upregulated in the 21d model group compared with those of 14d model group.Conclusion KDM2A is overexpressed in the lung tissues of BLM-induced pulmonary fibrosis in mice by regulating the expression of TGF-β1.
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