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[摘要] 目的 观察姜黄素对哮喘大鼠气道平滑肌细胞（ASMC）增殖及TGFβ1表达的影响。 方法 将24只雄性SD大鼠分为对照组、哮喘组和姜黄素组。以卵清白蛋白（OVA）致敏激发建立哮喘大鼠气道重塑模型，图像分析技术测定支气管壁厚度（Wat）和平滑肌厚度（Wam），免疫组织化学法（IHC）测定肺组织 TGFβ1 蛋白表达。酶联免疫吸附法（ELISA）测定细胞培养液中TGFβ1含量，实时定量 PCR（Real-time PCR）检测ASMC中TGFβ1mRNA表达，CCK-8分析ASMC增殖程度。 结果 姜黄素组与哮喘组相比，Wat及Wam厚度减少（P<0.01），仍高于对照组（P<0.01）；IHC显示：姜黄素组肺组织中 TGFβ1蛋白表达与哮喘组比较明显减少（P<0.01），仍高于对照组（P<0.01）。姜黄素组大鼠ASMC中TGFβ1和TGFβ1mRNA表达均降低，与对照组比较差异有统计学意义（P<0.01）；姜黄素组ASMC增殖明显受到抑制（P<0.01）。 结论 姜黄素能抑制哮喘大鼠ASMC增殖，其机制与降低TGFβ1的表达有关。
[关键词] 转化生长因子β1；大鼠；哮喘；气道重塑；姜黄素
[中图分类号] R285.5 [文献标识码] A [文章编号] 1673-9701（2016）19-0034-04
[Abstract] Objective To study the role of curcumin on proliferation mechanism of airway smooth musle cell （ASMC） and TGFβ1 expression in asthmatic rats. Methods A total of 24 Sprague-Dawley（SD） rats were sensitized and challenged by OVA to establish asthmatic model were divided into three groups with 8 rats in each group. The total bronchial wall thickness（Wat） and the airway smooth musle thickness（Wam） were measured by image analysis system. The TGFβ1 expressions were determined by immunohistochemistry. Vitro experiments were conducted to determine the direct effect of curcumin on TGFβ1 expression by ASMC. Real-time PCR was employed to detect the expression of TGFβ1 mRNA levels. TGFβ1 concentrations were determined by sandwich enzyme-linked immunosorbent assay（ELISA）. Cell Counting Kit-8（CCK-8） was used to detect ASMC proliferation. Results Wat and Wam of curcumin group were significantly thiner than asthma group （P<0.01）； but those were significantly thicker than control group （P<0.01）. The TGFβ1 expressions in the curcumin group was significantly lower than those of the asthmatic group（P<0.01）； but they were higher than the control group （P<0.01）. Compared to control group， curcumin decreased the protein expressions and mRNA expressions of TGFβ1 of ASMC （P<0.01）， inhibited ASMC proliferation （P<0.01）. Conclusion Curcumin could inhibit ASMC proliferation in asthmatic rats， which is associated withthe decrease of TGFβ1.
[Key words] TGFβ1； Rat； Asthma； Airway remodeling； Curcumin
支气管哮喘（简称哮喘）（bronchial asthma）的病理特征主要包括气道炎症和气道重塑。气道的结构细胞参与了气道重塑，其中平滑肌细胞（airway smooth muscle cell，ASMC）发生表型改变[1]，参与气道炎症的进程，对哮喘气道重塑发挥重要的作用[2]。转换生长因子（transforming growth factor，TGF）β是一类生物学功能复杂的细胞因子，其中TGFβ1具有促进平滑肌细胞的分化、生长与增殖，肺纤维化等[3]多种生物学效应。由于姜黄素在抗纤维化、抗炎等方面都发挥显著作用，近年来引起了越来越多学者的关注，尤其在哮喘方面的研究。姜黄素既往的研究主要以体内干预为主，但其对特定气道结构细胞培养水平如何发挥抗炎、抗纤维化的作用机制尚未阐明。本研究通过体外培养平滑肌细胞，探讨姜黄素对哮喘大鼠ASMC增殖及TGFβ1表达的影响，揭示姜黄素抑制哮喘气道重塑中ASMC增殖的机制，为其防治哮喘提供科学的实验依据。 1 材料与方法
1.1 实验动物
4～6周龄SPF级雄性Sprague-Dawley（SD）大鼠24只，体质量（100～120 g），购自上海斯莱克实验动物有限责任公司，动物许可证号：SCXK（沪）2008-003。
1.2 主要药品与试剂
姜黄素、卵清蛋白（OVA）购自美国sigma公司。TGFβ1小鼠抗大鼠单克隆抗体采购于美国Abcam公司，检测TGFβ1蛋白含量的ELISA试剂盒采购于美国R
[关键词] 转化生长因子β1；大鼠；哮喘；气道重塑；姜黄素
[中图分类号] R285.5 [文献标识码] A [文章编号] 1673-9701（2016）19-0034-04
[Abstract] Objective To study the role of curcumin on proliferation mechanism of airway smooth musle cell （ASMC） and TGFβ1 expression in asthmatic rats. Methods A total of 24 Sprague-Dawley（SD） rats were sensitized and challenged by OVA to establish asthmatic model were divided into three groups with 8 rats in each group. The total bronchial wall thickness（Wat） and the airway smooth musle thickness（Wam） were measured by image analysis system. The TGFβ1 expressions were determined by immunohistochemistry. Vitro experiments were conducted to determine the direct effect of curcumin on TGFβ1 expression by ASMC. Real-time PCR was employed to detect the expression of TGFβ1 mRNA levels. TGFβ1 concentrations were determined by sandwich enzyme-linked immunosorbent assay（ELISA）. Cell Counting Kit-8（CCK-8） was used to detect ASMC proliferation. Results Wat and Wam of curcumin group were significantly thiner than asthma group （P<0.01）； but those were significantly thicker than control group （P<0.01）. The TGFβ1 expressions in the curcumin group was significantly lower than those of the asthmatic group（P<0.01）； but they were higher than the control group （P<0.01）. Compared to control group， curcumin decreased the protein expressions and mRNA expressions of TGFβ1 of ASMC （P<0.01）， inhibited ASMC proliferation （P<0.01）. Conclusion Curcumin could inhibit ASMC proliferation in asthmatic rats， which is associated withthe decrease of TGFβ1.
[Key words] TGFβ1； Rat； Asthma； Airway remodeling； Curcumin
支气管哮喘（简称哮喘）（bronchial asthma）的病理特征主要包括气道炎症和气道重塑。气道的结构细胞参与了气道重塑，其中平滑肌细胞（airway smooth muscle cell，ASMC）发生表型改变[1]，参与气道炎症的进程，对哮喘气道重塑发挥重要的作用[2]。转换生长因子（transforming growth factor，TGF）β是一类生物学功能复杂的细胞因子，其中TGFβ1具有促进平滑肌细胞的分化、生长与增殖，肺纤维化等[3]多种生物学效应。由于姜黄素在抗纤维化、抗炎等方面都发挥显著作用，近年来引起了越来越多学者的关注，尤其在哮喘方面的研究。姜黄素既往的研究主要以体内干预为主，但其对特定气道结构细胞培养水平如何发挥抗炎、抗纤维化的作用机制尚未阐明。本研究通过体外培养平滑肌细胞，探讨姜黄素对哮喘大鼠ASMC增殖及TGFβ1表达的影响，揭示姜黄素抑制哮喘气道重塑中ASMC增殖的机制，为其防治哮喘提供科学的实验依据。 1 材料与方法
1.1 实验动物
4～6周龄SPF级雄性Sprague-Dawley（SD）大鼠24只，体质量（100～120 g），购自上海斯莱克实验动物有限责任公司，动物许可证号：SCXK（沪）2008-003。
1.2 主要药品与试剂
姜黄素、卵清蛋白（OVA）购自美国sigma公司。TGFβ1小鼠抗大鼠单克隆抗体采购于美国Abcam公司，检测TGFβ1蛋白含量的ELISA试剂盒采购于美国R