The aim of this study was to investigate whether transforming growth factor-h1 (TGF-h1) could induce alveolar epithelial to mesenchymal transition (EMT) in vitro. Alveolar epithelial cells (AECs) from SD rats were isolated by elastase cell dispersion and IgG panning. Expression of a-smooth muscle actin (a-SMA) was assayed using Western blotting and immunostaining analysis. Morphological changes, the markers of epithelial cell (E-cadherin), and stress fiber by actin reorganization were detected by an indirect immunostaining. The contents of collagen I were determined by spectrophotometry. The levels of endogenous TGF-h1 were measured with ELISA. Incubation of AECs with TGF-h1 (0.1～10ng/mL) induced abundant expression of a-SMA protein, and a-SMA expression in AECs reached a plateau when TGF-h1 was N 3ng/mL. Furthermore, we found that TGF-h1 (3ng/mL) exposure of AECs induced an authentic EMT characterized by abundant expression of a-smooth muscle actin, transformation of myofibroblastic morphology, increased formation of stress fiber by actin reorganization, and loss of epithelial marker E-cadherin. Meanwhile, significant increase in the levels of collagen I from 32.0 F 6.6mg/g in control to 98 F 10.8mg/g in TGF-h1-treated group was found over a 72h incubation period. Moreover, following stimulated by TGF-h1 (3ng/mL), a marked and time-dependent increase in endogenous TGF-h1 released from AECs was observed. At time points 72h, TGFh1 release mounted to 3451pg/ml, which was much enough to induce EMT in vitro. These results demonstrated that AECs, under stimulation of TGF-h1, underwent a conversion process into myofibroblasts in vitro.

本研究目的旨在观察PDE IV 抑制剂咯利普兰对沙丁胺醇（SB）耐受性的影响。用豚鼠离体气管标本，预先接触SB1μmol•L-1，1h可使SB对抗乙酰甲胆碱气管收缩作用的剂量反应曲线右移5倍，最大松弛率下降30%，形成SB气管松弛作用的耐受性。磷酸二酯酶（PDE）IV抑制剂咯利普兰能翻转SB气管松弛作用的耐受性，但PDE Ⅲ抑制剂氰胍哒嗪则不能。虽然蛋白合成抑制剂环己酰亚胺对SB耐受性无预防作用，但上述结果仍提示SB气管松弛作用的耐受性可能与PDEIV活性的增高有关。

探讨隐孔菌发酵物（Cryp top orus volvatus ferment substance, CVFS）对中性粒细胞释放白三烯的影响。方法：给大鼠腹腔注射糖原诱导中性粒细胞聚集，16h后收集腹腔灌洗液，分离中性粒细胞，以calcium ionopho re A 23187刺激离体中性粒细胞释放白三烯，再用HPLC法测定中性粒细胞中白三烯（LT）B4、L TC4和LTD4的含量。结果：CVFS 0.25、1、4mg•L-1呈浓度依赖抑制中性粒细胞释放LTB4、LTC4和LTD4，对LTB4抑制率分别为27.4%、54.2%和78.8%；对LTC4抑制率分别为65.1%、74.3 和79.0%；对LTD4抑制率分别为5516%、60.9%和72.8%。与生理盐水对照组比较有显著性差异（P<0.05）。结论：CVFS抑制中性粒细胞释放LTB4、LTC4和LTD4可能是其抗炎平喘作用机制之一。

探讨致敏后重复抗原攻击大鼠肺组织CGMP-PDE、CAMP-PDE活性变化。方法：HPLC法测定致敏后重复抗原攻击的大鼠（抗原攻击组）、致敏不攻击的大鼠（致敏组）和正常大鼠（正常组）肺组织CGMP-PDE、CAMP-PDE活性。结果：抗原攻击组肺组织CGMP-PDE、CAMP-PDE活性分别为798.76-529.23、597.50-220.75nmol•g-1protein•min-1，其中抗原攻击组大鼠肺组织CGM P-PDE活性较正常组明显升高（P<0.05），抗原攻击组大鼠肺组织CAMP-PDE活性较致敏组和正常组均明显升高（P<0.05）。结论：大鼠哮喘发病过程中肺组织CGMP-PDE、CAMP-PDE活性明显升高。

目的研究环孢素A（CsA）气雾吸入对抗原诱导的大鼠过敏性气道炎症的作用。方法用卵白蛋白（OA）致敏大鼠，2周后气雾吸入CsA（5，10，20 g•L-1），每天1次，连续7d。大鼠致敏后d20和d21用OA（10g•L-1，每天1次）攻击，观察第2次OA攻击24h后支气管肺泡灌洗液及外周血中嗜酸性粒细胞的数量和支气管肺组织病理学改变情况，测定支气管肺泡灌洗液中TNF-α含量。结果CsA气雾吸入能明显降低支气管肺泡灌洗液及外周血中嗜酸性粒细胞的数量，减轻肺组织中炎症细胞特别是嗜酸性粒细胞的浸润，减轻组织水肿及上皮损伤等气道炎症状况，降低支气管肺泡灌洗液中TNF-α含量。结论CsA气雾吸入对大鼠过敏性气道炎症具有抑制作用，其作用机制与细胞因子TNF-α释放减少有关。