Wang Xun, Li Ji-qiong, Chen Lei, Wang Hai-xia, Sun Bei-bei, Wang Tao, Xu Dan, Wen Fu-qiang
Division of Pulmonary Diseases, National Key Laboratory of Biotherapy of Human Diseases, Department of Respiratory Medicine, West China Hospital of Sichuan University, Chengdu 610041, China.
Zhonghua Yi Xue Za Zhi. 2008 Nov 18;88(42):2988-93.
To examine the effects of YM976, a phosphodiesterase (PDE)4 inhibitor, on mucin hypersecretion of airway stimulated with acrolein.
Forty-eight SD rat were randomly divided into 6 equal groups. Twenty-four rats underwent gastric perfusion of YM976 0.5, 1.5, or 4.5 mgxkg(-1)xd(-1) (n=8 for each group) and then underwent aerosol inhalation of 3.0 ppm acrolein 6 hours per day for 12 days so as to establish rat models of airway mucus hypersecretion. Eight rats underwent aerosol inhalation of 3.0 ppm acrolein only for 12 days to establish rat models of airway mucus hypersecretion without any drug intervention (acrolein model group). Eight rats underwent gastric perfusion of 5% methylcellulose and then acrolein inhalation for 12 days (methylcellulose group), and another 8 rats underwent gastric perfusion of YM976 4.5 mg/kg once a day and then inhalation of normal saline for 12 days (YM976 control group). On the 13th day the rats were killed. Bronchoalveolar lavage fluid (BALF) was acquired for cell count, and ELISA was used to detect the levels of tumor necrosis factor (TNF)-alpha and cytokine-induced neutrophil chemoattractant (CINC)-1 in the BALF. The mucin level in the airway mucous membrane was detected by alcian blue(AB)/periodic acid-Schiff(PAS) method. Muc5ac protein was measured by immunohistochemical staining and Western-blotting, and Muc5ac mRNA was detected by RT-PCR.
The percentages of positive AB/PAS staining of the acrolein model group and the 3 YM976 groups (0.5, 1.5, and 4.5 mgxkg(-1)xd(-1)) were 23.65+/-2.86, 22.63+/-2.12, 16.34+/-1.72, and 5.03+/-0.72 respectively. The integral optical density (IOD) levels of Muc5ac immunohistochemical staining of the methylcellulose group and the 3 YM976 groups were 0.54+/-0.05, 0.49+/-0.06, 0.32+/-0.04, and 0.22+/-0.04 respectively. The IOD of Muc5ac protein expression by Western-blotting of the methylcellulose group and the 3 YM976 groups were 1.177+/-0.190, 0.806+/-0.180, 0.303+/-0.061, and 0.134+/-0.035 respectively. The IOD of Muc5ac mRNA expression by RT-PCR of the methylcellulose group and the 3 YM976 groups were 0.246+/-0.021, 0.223+/-0.020, 0.161+/-0.012, and 0.097+/-0.011 respectively. The TNF-alpha and CINC-1 levels in the BALF of the methylcellulose group and the 3 YM976 groups were (96.77+/-2.31), (87.65+/-2.75), (73.56+/-2.62), and (52.23+/-2.79) microg/L respectively, and (145.75+/-4.43), (139.73+/-5.51), (95.34+/-5.13), and (65.74+/-5.62) microg/L respectively, all significantly higher than those of the normal control group [4.38+/-0.32, 0.12+/-0.02, 0.058+/-0.024, 0.061+/-0.006, (18.23+/-2.32) microg/L and (33.56+/-3.43) microg/L respectively, all P<0.05], but those of the YM976 1.5 mgxkg(-1)xd(-1) and 4.5 mgxkg(-1)xd(-1) groups being significantly lower than those of the methylcellulose group (both P<0.05).
YM976 inhibits airway inflammatory response and airway mucin hypersecretion induced by acrolein.
研究磷酸二酯酶（PDE）4抑制剂YM976对丙烯醛刺激气道黏液高分泌的影响。
48只SD大鼠随机分为6组，每组8只。24只大鼠分别经胃灌注0.5、1.5或4.5mg·kg⁻¹·d⁻¹的YM976，然后每天6小时雾化吸入3.0ppm丙烯醛，共12天，以建立气道黏液高分泌大鼠模型。8只大鼠仅雾化吸入3.0ppm丙烯醛12天，建立无药物干预的气道黏液高分泌大鼠模型（丙烯醛模型组）。8只大鼠经胃灌注5%甲基纤维素后吸入丙烯醛12天（甲基纤维素组），另8只大鼠每天经胃灌注4.5mg/kg的YM976，然后吸入生理盐水12天（YM976对照组）。第13天处死大鼠，获取支气管肺泡灌洗液（BALF）进行细胞计数，采用酶联免疫吸附测定（ELISA）法检测BALF中肿瘤坏死因子（TNF）-α和细胞因子诱导的中性粒细胞趋化因子（CINC）-1水平。采用阿尔辛蓝（AB）/过碘酸-希夫（PAS）法检测气道黏膜中的黏蛋白水平。采用免疫组织化学染色和蛋白质印迹法检测Muc5ac蛋白，采用逆转录-聚合酶链反应（RT-PCR）法检测Muc5ac mRNA。
丙烯醛模型组及3个YM976组（0.5、1.5和4.5mg·kg⁻¹·d⁻¹）AB/PAS染色阳性率分别为23.65±2.86、22.63±2.12、16.34±1.72和5.03±0.72。甲基纤维素组及3个YM976组Muc5ac免疫组织化学染色的积分光密度（IOD）水平分别为0.54±0.05、0.49±0.06、0.32±0.04和0.22±0.04。甲基纤维素组及3个YM976组蛋白质印迹法检测Muc5ac蛋白表达的IOD分别为1.177±0.190、0.806±0.180、0.303±0.061和0.134±0.035。甲基纤维素组及3个YM976组RT-PCR法检测Muc5ac mRNA表达的IOD分别为0.246±0.021、0.223±0.020、0.161±0.012和0.097±0.011。甲基纤维素组及3个YM976组BALF中TNF-α水平分别为（96.77±2.31）、（87.65±2.75）、（73.56±2.62）和（52.23±
