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目的:观察抑制环磷酸腺苷(cAMP)直接激活的交换蛋白(Epac)对慢性哮喘小鼠模型气道重塑的影响。方法:18只雌性BALB/c小鼠随机分为正常对照组(PBS组)、哮喘组(OVA组)、Epac抑制剂组(ESI-09组),每组6只。3组小鼠均给予卵清蛋白(OVA)致敏,PBS组给予PBS激发,OVA组和ESI-09组给予OVA激发,其中ESI-09组于每次激发前30 min给予腹腔注射ESI-09(溶于DMSO)处理,PBS组和OVA组对应给予等浓度DMSO处理。末次激发24h后,取小鼠肺组织固定、切片行PAS、Masson三色染色及α-平滑肌肌动蛋白(α-SMA)免疫组织化学检测,以便分别对气道杯状细胞增生、胶原沉积及平滑肌细胞增生肥大等情况进行评估,结果以杯状细胞(PAS+)、胶原(Masson+)及平滑肌细胞(α-SMA+)着色面积/支气管基底膜周长(μm2/μm)表示。结果:OVA组和ESI-09组杯状细胞的增生均显著高于PBS组[3.05±0.12、11.45±1.28比0.00±0.00(PBS组未见PAS+区域),均P<0.01],且ESI-09组显著高于OVA组(P<0.05);Masson染色显示,胶原的沉积OVA组和ESI-09组均显著高于PBS组(3.40±0.35、5.28±0.58比1.10±0.11,均P<0.01),且ESI-09组显著高于OVA组(P<0.01);α-SMA免疫组化可见OVA组α-SMA的表达显著高于PBS组(3.90±0.23比2.48±0.39,P<0.05),且ESI-09组(5.55±0.59)又显著高于OVA组(P<0.05)。结论:抑制Epac明显加重了慢性哮喘小鼠模型气道重塑,因此cAMP-Epac信号通路可能对慢性哮喘小鼠气道重塑起保护作用。
AIM: To observe the effect of Epac, an inhibitor of cAMP activation, on airway remodeling in a mouse model of chronic asthma. Methods: Eighteen female BALB / c mice were randomly divided into normal control group (PBS group), asthma group (OVA group) and Epac inhibitor group (ESI-09 group), with 6 mice in each group. The OVA group and OVA group were given OVA challenge. The ESI-09 group was given intraperitoneal injection of ESI-09 30 min before each challenge (Dissolved in DMSO), PBS group and OVA group corresponding to the same concentration of DMSO treatment. Twenty-four hours after the last challenge, the lung tissues of the mice were fixed and examined by PAS, Masson’s trichrome staining and α-smooth muscle actin (α-SMA) immunohistochemistry in order to detect the proliferation of airway goblet cells, collagen deposition and Smooth muscle cell hyperplasia and hypertrophy were evaluated. The results were expressed as PAS +, Masson + and α-SMA + staining area / bronchial basement membrane perimeter (μm2 / μm). Results: The hyperplasia of goblet cells in OVA group and ESI-09 group was significantly higher than that in PBS group [3.05 ± 0.12, 11.45 ± 1.28 vs 0.00 ± 0.00 (PAS + region was not observed in PBS group, both P <0.01] Masson staining showed that collagen deposition in OVA group and ESI-09 group were significantly higher than those in PBS group (3.40 ± 0.35, 5.28 ± 0.58 vs 1.10 ± 0.11, P <0.01, respectively) ), And the expression of α-SMA in OVA group was significantly higher than that in OVA group (3.90 ± 0.23 vs 2.48 ± 0.39, P <0.05) , And ESI-09 group (5.55 ± 0.59) was significantly higher than OVA group (P <0.05). Conclusion: Inhibition of Epac significantly aggravates airway remodeling in a chronic asthmatic mouse model. Therefore, cAMP-Epac signaling pathway may play a protective role in airway remodeling in chronic asthmatic mice.