Ma J Y, Cheng Y H, Barger M W, Xin W F, Lewis D M, Castranova V, Ma J K
School of Pharmacy, West Virginia University, Morgantown 26506, USA.
Exp Lung Res. 1995 Sep-Oct;21(5):771-90. doi: 10.3109/01902149509050841.
Bis-basic ethers of fluorene and fluoren-9-substituted derivatives such as tilorone have been reported to inhibit silica-induced fibrosis in rats. The potential antifibrotic potency of 2,7-bis(diethylamino)ethoxy fluorene (F-9-H,H), fluorenone (F-9-one), fluorenoxime (F-9-oxime), and fluorenol (F-9-ol) was F-9-oxime > F-9-one approximately F-9-H,H >> F-9-ol. Since the release of reactive oxygen species and growth factors from alveolar macrophages (AM) in response to silica exposure has been linked to the development of pulmonary fibrosis, the present study was carried out to determine the inhibitory effects of these compounds on rat AM activity in vitro. The following parameters were monitored: (1) cellular viability; (2) zymosan-induced respiratory burst activity (superoxide and hydrogen peroxide release, chemiluminescence, and oxygen consumption) of AM; (3) drug binding to AM; and (4) lipopolysaccharide (LPS)-stimulated interleukin-1 (IL-1) release from AM. The bis-basic ethers, at 40 microM, did not affect cell viability when incubated with AM for 30 min, but significantly inhibited zymosan-induced macrophage respiratory burst activity. The inhibitory effect of these agents was F-9-oxime > F-9-one approximately F-9-H,H >> F-9-ol. Binding of these drugs to AM was time and dose dependent, and exhibited the following binding affinity: F-9-oxime > F-9-one > F-9-H,H > F-9-ol. F-9-oxime was shown to inhibit LPS-stimulated IL-1 release by AM in a dose-dependent manner. This inhibition of IL-1 release by AM cannot be explained as a decrease in viability. In addition, these drugs were also shown to impair human fibroblast proliferation in response to serum stimuli without impairing cell viability. These results indicate a positive correlation between drug binding to AM or other cell types and their inhibitory effects on cellular activities including oxygen consumption, superoxide release, hydrogen peroxide secretion, chemiluminescence, IL-1 release, and proliferation. The ability of these bis-basic ethers to modify AM and fibroblast functions in vitro suggests that further investigation of their reported antifibrotic potency in vivo is warranted.
芴及芴-9-取代衍生物（如梯洛龙）的双碱性醚类已被报道可抑制大鼠二氧化硅诱导的纤维化。2,7-双（二乙氨基）乙氧基芴（F-9-H,H）、芴酮（F-9-one）、芴肟（F-9-oxime）和芴醇（F-9-ol）的潜在抗纤维化效力为F-9-oxime > F-9-one ≈ F-9-H,H >> F-9-ol。由于肺泡巨噬细胞（AM）在接触二氧化硅后释放活性氧和生长因子与肺纤维化的发展有关，因此开展本研究以确定这些化合物对大鼠AM体外活性的抑制作用。监测了以下参数：（1）细胞活力；（2）AM的酵母聚糖诱导的呼吸爆发活性（超氧化物和过氧化氢释放、化学发光及耗氧量）；（3）药物与AM的结合；（4）脂多糖（LPS）刺激的AM白细胞介素-1（IL-1）释放。双碱性醚类在40微摩尔浓度下与AM孵育30分钟时不影响细胞活力，但显著抑制酵母聚糖诱导的巨噬细胞呼吸爆发活性。这些药物的抑制作用为F-9-oxime > F-9-one ≈ F-9-H,H >> F-9-ol。这些药物与AM的结合具有时间和剂量依赖性，并表现出以下结合亲和力：F-9-oxime > F-9-one > F-9-H,H > F-9-ol。F-9-oxime被证明以剂量依赖性方式抑制LPS刺激的AM释放IL-1。AM对IL-1释放的这种抑制不能解释为活力下降。此外，这些药物还被证明可损害人成纤维细胞对血清刺激的增殖反应，而不损害细胞活力。这些结果表明药物与AM或其他细胞类型的结合与其对包括耗氧量、超氧化物释放、过氧化氢分泌、化学发光、IL-1释放和增殖在内的细胞活性的抑制作用之间存在正相关。这些双碱性醚类在体外改变AM和成纤维细胞功能的能力表明有必要进一步研究它们在体内所报道的抗纤维化效力。
