目的：比较RR-福莫特罗和rac-福莫特罗对人支气管舒张作用。方法：将内径2～4mm、长15mm的人支气管螺旋条置入持续供氧的浴槽内，静息张力为1g，以蓄积给药法测定RR-福莫特罗（10 pmol•L-1～3.2 Lmol•L-1）和rac-福莫特罗（10 pmol•L-1～3.2 Lmol•L-1）在静息情况下，及氨甲酰胆碱（10 Lmo l•L-1）或组胺（100 Lmol•L-1）引起收缩情况下的张力改变。结果：RR-福莫特罗在静息状况下支气管舒张作用强于rac-福莫特罗（P<0.05）。RR-福莫特罗和rac-福莫特罗对抗由氨甲酰胆碱和组胺引起的收缩，且RR-福莫特罗的对抗作用强于rac-福莫特罗（P<0.05）。结论：RR-福莫特罗对人支气管的舒张作用强于rac-福莫特罗。

To explore the changes of leukotrienes (LT) in cerebral cortex and lung tissues in ovalbumin-induced rat asthma model and effects of different anti-asthma drugs on the changes. METHODS: Aerosol antigen-induced changes of inflammation in bronchoalveolar lavage fluids (BALF), pulmonary and brain histologic section in sensitized rats were investigated. Changes of LTB4 and LTC4 in lung and cerebral cortex homogenates were analyzed by reverse-phase high performance liquid chromatography (RP-HPLC). RESULTS: The number of inflammatory cells in BALF and the score of lung and brain histological examination from antigen- challenged rats were significantly higher than that from control group (P<0.05). Dexamethason (DXM, 0.5mg/kg, ip) and ketotifen fumarate (KF, 5mg/kg, ig) markedly reduced total leukocyte number in BALF, and inhibited eosinophil accumulation, reduced the infiltration of eosinophils, and improved mucous edema and epithelial lesion of bronchi and bronchioles. In addition, RP-HPLC results shown LTB4 in lung and cerebral cortex homogenates were increased in antigenchallenged rats [(4.1

To study the inductive expression of human phosphodiesterase 4A (hPDE4A) in yeast cell GL62 and investigate the inhibitory effects of theophylline, rolipram, and acetamide-45 on PDE4A activity of the expressed product in yeast cell GL62. METHODS: Yeast cell GL62 were transfected with human PDE4A gene cloned in the expression plasmid p138NB. Expression was induced by adding CuSO4 to a final concentration of 150μmol/L, and the expressed product was extracted. The activity of PDE4A was detected by HPLC. RESULTS: Yeast cell GL62 expressed a certain protein at CuSO4 150μmol/L, the size of the expressed product was between 62 kDa and 83 kDa, the activity of PDE4A of the expressed product at 3 h was in maximum (188±23)μmol×g-1×min-1, and the Km was (17.7±2.6) mmol/L. Theophylline, rolipram, and acetamide-45 could inhibit the activity of PDE4A extracted from yeast cell GL62. The IC50 (95% confidence limits) of theophylline, rolipram, and acetamide-45 were 1642 (989-2727), 4.58 (3.45-6.08), and 275 (170-444) mmol/L respectively. CONCLUSION: PDE4A expressed in yeast cell GL62 is biologically active. Theophylline, rolipram, and acetamide-45 can inhibit the PDE4A activity. The expressed product in yeast cell GL62 may be used in the research work of PDE4 and its inhibitors.

目的：评价环孢素A气雾给药对豚鼠哮喘模型的药效。方法：用乙酰胆碱（ACh）或组胺诱导抗原攻击后的致敏豚鼠气道阻力pC200、支气管肺泡灌洗液（BALF）和肺组织切片中的嗜酸性粒细胞（EOS）变化观察环孢素A气雾给药后的抗气道高反应性和炎症作用。结果：环孢素AlOg•L-1、20g•L-1气雾给药和地塞米松（0.5mg-1kg ip）增加PC200值，能预防ACh或组胺引起的气道高反应性。环孢素A5g-L-1对组胺引起的气道高反应性也有作用，对Ach不显著．环孢素A10g•L-1、20g•L-1气雾给药能明显减少BAIF中的EOS浸润。与溶媒组比较，地塞米松0.5mg•kg-1增加了BALF中的中性粒细胞数日，与三组环孢素A比较有显著差异。在肺组织学研究中，环孢素A20g•L-1和地塞米松0.5mg•kg-1可抑制支气管和细支气管上皮和上皮表面结缔组织的EOS浸润。结论：环孢素A气雾吸入给药能明显对抗致敏豚鼠气道高反应性和炎症反应，为其治疗哮喘提供了一个可选择的给药途径。

目的：探讨致敏大鼠抗原攻击后脑皮层和肺气道中干扰素-γ（-IFN-γ）和白介素-4（IL-4）出现的相关性变化。方法：观察致敏大鼠吸入抗原诱导的支气管肺灌洗液（BAIF）和肺组织切片炎症变化，用ELISA法测定BALF和脑皮层-IFN-γ和JL-4水下变化。结果：抗原攻击组BAIF中的炎症细胞数目明显高于对照未攻击组（p<0.05）。地塞米松（DXM，0.5mg/kg，ip）明显减少BAIF中的白细胞总数，几乎完全抑制嗜酸性粒细胞（EOS）和淋巴细胞的聚集，但增加中性粒细胞数目。抗原攻击组的组织学检查积分（EOS浸润、粘膜水肿和上皮损伤）也明显高于对照未攻击组（P<O.05）。 DXM （0.5mg/kg，ip）减少支气管和细支气管的：EOS数目，改善粘膜水肿和上皮损伤。致敏人鼠抗原攻击后。BAIJF中的IFN，7水平降低伴随IL-4升高导致了IFN-γ/IL-4比例下降与此同时，脑皮层匀浆中也出现相似的改变。DXM（0.5mg/kg，Ip）能反转BALF和脑皮层匀浆中的IFN-γ/IL-4比例下降。结论：致敏大鼠抗原攻击后脑皮层和肺气道中的IFN-γ/IL-4出现相关性变化。

To compare the bronchodilating and antiinflammatory effects of oral racemic formoterol (rac-FMT) and (R,R)-formoterol (R,R-FMT). METHODS: The changes of lung resistance (RL), dynamic lung compliance (Cdyn), and the accumulation of inflammatory cells in bronchoalveolar lavage fluids (BALF) induced by ovalbumin aerosol in sensitized guinea pigs and mice were investigated in vivo. RESULTS: Mean value increase of RL and mean value reduction of Cdyn from 1 to 30 min after antigen challenge were up to 101%

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.

目的：研究硝普钠（SNP）与异丙肾上腺素（ISO）对犬气管平滑肌松弛作用是否存在协同关系，并分析协同的机制。方法：采用放射免疫分析法测定犬气管平滑肌环核苷酸量，并用磷酸二酯酶（PDE）同工酶抑制剂为工具分析SNP与ISO相互作用的机制。结果：SNP（1～100Lmol•L-1）增强ISO（30Lmol•L-1）对3Lmol•L-1乙酰甲胆碱预收缩的犬气管平滑肌的松弛作用。SNP（100Lmol•L-1） 与ISO（30Lmol•L-1）对犬气管平滑肌松弛的协同作用伴有气管平滑肌中CAMP积聚比SNP与ISO单独时分别增加112倍与014倍。PDEÍ 抑制剂扎普司特（3 Lmol•L-1）使SNP的气管平滑肌CGM P积聚由214倍增加到416倍，可进一步增加气管松弛作用112倍。合并PDEË抑制剂氰胍哒嗪（30Lmol•L-1）和PDEÌ抑制剂咯利普兰（30Lmol•L-1）或合并SNP（100Lmol•L-1） 和咯利普兰（30Lmol•L-1）均能进一步增强ISO的气管cAMP积聚与松弛作用。结论：SNP是通过气管平滑肌cGMP含量增加而抑制PDEË，导致SNP与ISO对气管松弛的协同作用。

human PDE4A was improved by ammonium sulfate fractionation, DEAE Sephadex A-50 chromatography, and Sephadex G-100 chromatography. The activity of PDE4A was measured by high performance liquid chromatography (HPLC). RESULTS: Induced PDE4A activity expressed in crude yeast cell GL62 supernatant and pellet was (340±21) nmol•g-1•min-1 and (250±25) nmol•g-1•min-1 respectively. The specific activity of recombinant PDE4A in supernatant was improved 6.4 fold. Ciclamilast, piclamilast, and rolipram could inhibit PDE4A activity. The IC50 values (95% confidence limits) of ciclamilast, piclamilast, and rolipram were 1.27 (0.84-1.91), 66.4 (33.3-132.2), and 3.73 (2.51-5.53) μmol/L respectively. Zaprinast, aspirin, and indomethacin had no obvious inhibitory effect on PDE4A activity. CONCLUSION: The specific activity of PDE4A expressed in yeast cell GL62 can be improved by ammonium sulfate fractionation, DEAE Sephadex A-50 chromatography, and Sephadex G-100 chromatography. Ciclamilast, piclamilast, and rolipram can inhibit PDE4A activity while zaprinast, aspirin, and indomethacin have no obvious inhibitory effect on PDE4A activity. Human PDE4A expressed in GL62 might be useful in the research and screening of new selective PDE4 inhibitors.

上皮完整或去上皮的豚鼠离体气管以300Lmol•L-1硝普钠（SNP）预处理1h，使SNP对抗乙醋甲胆碱气管收缩作用的剂量反应曲线右移1.3-1.5倍，最大舒张率下降41%-58%，形成SNP气管松弛作用的耐受性82溴环鸟苷酸可模拟SNP在豚鼠离体气管形成的SNP耐受性，谷胱甘肽（1mmol•L-1）及环核苷酸磷酸二酯酶（PDE）V抑制剂扎普司特（30Lmol•L-1）均可部分翻转SNP的气管松弛作用耐受性，而蛋白合成抑制剂环己酰亚胺（10Lmol•L-1）对SNP的耐受性无预防作用。结果表明SNP可产生豚鼠离体气管松弛耐受性，这可能是由于气管平滑肌中环鸟苷酸（CGMP）积聚而下调鸟苷酸环化酶（GC）活性和上调PDEÍ活性。