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香烟烟雾刺激培养的牛支气管上皮细胞释放中性粒细胞趋化活性。

Cigarette smoke stimulates release of neutrophil chemotactic activity from cultured bovine bronchial epithelial cells.

作者信息

Shoji S, Ertl R F, Koyama S, Robbins R, Leikauf G, Von Essen S, Rennard S I

机构信息

University of Nebraska Medical Center, Omaha, USA.

出版信息

Clin Sci (Lond). 1995 Mar;88(3):337-44. doi: 10.1042/cs0880337.

Abstract
  1. Recruitment of neutrophils into the airway is a prominent feature of chronic bronchitis, a syndrome often associated with exposure to cigarette smoke. Since bronchial epithelial cells are the 'first' lung cells to come into contact with smoke, these cells may be responsible for neutrophil recruitment into the airway by release of neutrophil chemotactic activity in response to cigarette smoke. 2. To evaluate this hypothesis, we prepared bovine bronchial epithelial cells and measured their ability to release neutrophil chemotactic activity following exposure to cigarette smoke. Bronchial epithelial cells were prepared by overnight digestion with protease, filtered through 100-microns Nitex mesh and resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum and antibiotics and cultured at 2 x 10(6) cells in 2 ml of medium in 35-mm culture dishes. After 4 days, dishes were rinsed and refed with medium without fetal calf serum and incubated with and without 1:10 diluted smoke extract for 6 h at 37 degrees C. The neutrophil chemotactic activity of the supernatant fluids was measured by the blindwell chamber technique. 3. Cigarette smoke itself added to medium did not stimulate chemotaxis. In contrast, cigarette smoke did stimulate the release of neutrophil chemotactic activity from bovine bronchial epithelial cells [15 +/- 3 (control) versus 74 +/- 5 (smoke), P < 0.01]. 4. This neutrophil chemotactic activity was dialysable, pepsin and acid stable, heat sensitive and lipid extractable. Sephadex G-75 column chromatography demonstrated two peaks of neutrophil chemotactic activity.(ABSTRACT TRUNCATED AT 250 WORDS)
摘要
  1. 中性粒细胞募集到气道是慢性支气管炎的一个显著特征,慢性支气管炎是一种常与接触香烟烟雾相关的综合征。由于支气管上皮细胞是肺部最先接触烟雾的细胞,这些细胞可能通过释放中性粒细胞趋化活性来促使中性粒细胞募集到气道,以应对香烟烟雾。2. 为了评估这一假设,我们制备了牛支气管上皮细胞,并测量了它们在接触香烟烟雾后释放中性粒细胞趋化活性的能力。通过用蛋白酶过夜消化来制备支气管上皮细胞,将其通过100微米的尼泰克斯网过滤,并重悬于补充有10%胎牛血清和抗生素的杜尔贝科改良伊格尔培养基中,以2×10⁶个细胞/2毫升培养基的密度接种于35毫米培养皿中培养。4天后,冲洗培养皿,更换为不含胎牛血清的培养基,并在37℃下分别与1:10稀释的烟雾提取物孵育和不孵育6小时。通过盲孔室技术测量上清液的中性粒细胞趋化活性。3. 添加到培养基中的香烟烟雾本身并不会刺激趋化作用。相反,香烟烟雾确实能刺激牛支气管上皮细胞释放中性粒细胞趋化活性[对照组为15±3,烟雾组为74±5,P<0.01]。4. 这种中性粒细胞趋化活性是可透析的,对胃蛋白酶和酸稳定,对热敏感且可被脂质提取。葡聚糖G-75柱色谱显示出两个中性粒细胞趋化活性峰。(摘要截断于250字)

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