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The effect and the mechanism of Roxithromycin on apoptosis of airway smooth muscle cells from asthmatic rats in vitro |
1.Intensive Care Unit, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325027; 2.Department of Respiratory Medicine, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325027
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Cite this article: |
WU Haiya1,DAI Yuanrong2,YING Binyu1.. The effect and the mechanism of Roxithromycin on apoptosis of airway smooth muscle cells from asthmatic rats in vitro[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2015, 45(4): 252-.
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Abstract Objective: To observe the effects of Roxithromycin on apoptosis of airway smooth muscle cells (ASMCs) in vitro and the role of mitochondrial pathway in apoptosis of ASMCs. Methods: ASMCs from asthmatic rats were cultured in vitro and intervened with various concentrations of Roxithromycin for 48 hours. Early apoptotic cells were detected through flow cytometry (FCM) after stained with Annexin V/PI; Alteration of mitochondrial membrane potential (ΔΨm) was also examined by FCM after stained with JC-1; Cytochrome C (Cyt-c)
both in mitochondria and cytoplasm was analyzed by Western Blot. Results: Roxithromycin could induce the apoptotic death of ASMCs in vitro: Early apoptotic rate in experimental group (R10, R25, R50, R100) after 48 h intervention was (4.6±1.9)%, (5.8±2.9)%, (12.0±5.6)% and (26.9±11.1)%, respectively, compared to control (2.9±1.7)%. And there was a significant difference (x2=13.30, P<0.01) between the group R100 and the control.RXM induced loss of ΔΨm: The proportion of low ΔΨm cells of the experimental group (R10, R25, R50, R100) was (27.88±13.10)%, (40.35±9.19)%, (48.40±14.15)% and (52.90±15.88)%, respectively, compared to the control group (19.78±9.85)%. The indexes of group R100 and R50 were significantly lower than that of group R0 and R10 (P<0.05 or 0.01). The difference between group R25 and R0 was also statistically significance (P<0.05).But there was no statistical difference between group R10 and R0. Roxithromycin could induce Cyt-c release from mitochondria into the cytosol: The content of cytosolic Cyt-c in group R10, R25, R50, R100 was respectively (0.87±0.19), (0.97±0.17), (1.01±0.12), (1.14±0.07), higher than that in the control group (0.67±0.12). There was a significant difference between the group R100 and R0 (x2=9.075, P<0.05). Conclusion: Roxithromycin can induce apoptosis of ASMCs in vitro via activating mitochondrial pathway.
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Received: 07 August 2014
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[1] Labro MT. Anti-inflammatory activity of macrolides: a new therapeutic potential?[J]. J Antimicrob Chemoth, 1998, 41 (Suppl 2): 37-46.
[2] Koh Y, Lee M, Sun Y, et al. Effect of roxithromycin on airway responsiveness in children with bronchiectasis: a doubleblind, placebo-controlled study[J]. Eur Respir J, 1997, 10(5):994-999.
[3] Shimizu T, Kato M, Mochizuki H, et al. Roxithromycin reduces the degree of bronchial hyperresponsiveness in children with asthma[J]. Chest, 1994, 106(2): 458-461.
[4] Tada M , Saqara H, Okada S, et al. Effects of roxithromycin on a guinea pig model for chronic asthma [J]. Jpn J Antibiot, 2004, 57(Suppl A): 76-78.
[5] Tomita H, Osanai T, Toki T, et al. Roxithromycin is an inhibitor of human coronary artery smooth muscle cells proliferation: a potential ability to prevent coronary heart disease [J]. Atherosclerosis, 2005, 182(1): 87-95.
[6] Stamatiou R, Boukas K, Paraskeva E, et al. Azithromycin reduces the viability of human bronchial smooth muscle cells [J]. J Antibiot, 2010, 63(2): 71-75.
[7] Stamatiou R, Paraskeva E, Boukas K, et al. Azithromycin has an antiproliferative and autophagic effect on airway smooth muscle cells[J]. Eur Respir J, 2009, 34(3): 721-730.
[8] 吴斌, 王秀芬, 戴元荣. 罗红霉素对支气管哮喘大鼠气道平滑肌细胞凋亡及P27KIP-1蛋白表达的影响[J]. 中华结核与呼吸杂志, 2010, 33(12): 937-939.
[9] Palmans E, Kips JC, Pauwels RA. Prolonged allergen exposure induces structural airway changes in sensitized rats[J].Am J Resp Crit Care, 2000, 161(2): 627-635.
[10] 吴海亚, 戴元荣, 尹娟. 改良组织贴块法培养大鼠气道平滑肌细胞[J]. 温州医学院学报, 2010, 40(6): 571-573.
[11] 卓致远, 黄茂, 崔学范, 等. 组织贴块法培养小鼠气道平滑肌细胞[J]. 中国组织化学与细胞化学杂志, 2007, 16(2): 247-250.
[12] Ly JD, Grubb DR, Lawen A. The mitochondrial membrane potential (Δψm) in apoptosis; an update[J]. Apoptosis, 2003,8(2): 115-128.
[13] Hirst SJ. Regulation of airway smooth muscle cell immunomodulatory function: role in asthma [J]. Resp Physiol Neurobi, 2003, 137(2): 309-326.
[14] Johnson PRA, Roth M, Tamm M, et al. Airway smooth muscle cell proliferation is increased in asthma[J]. Am J Resp Crit Care, 2001, 164(3): 474-477.
[15] Glander HJ. Binding of annexin V to plasma membranes of human spermatozoa: a rapid assay for detection of membrane changes after cryostorage[J]. Mol Hum Reprod, 1999, 5(2):109-115.
[16] Vermes I, Haanen C, Steffens-Nakken H, et al. A novel assay for apoptosis flow cytometric detection of phosphatidyl-serine expression on early apoptotic cells using fluorescein labelled annexin V [J]. J Immunol Methods, 1995, 184(1):39-51.
[17] Gulbins E, Dreschers S, Bock J. Role of mitochondria in apoptosis[J]. Exp Physiol, 2003, 88(1): 85-90.
[18] Garrido C, Galluzzi L, Brunet M, et al. Mechanisms of cytochrome c release from mitochondria[J]. Cell Death Differ,2006, 13(9): 1423-1433.
[19] Susin SA, Lorenzo HK, Zamzami N, et al. Molecular characterization of mitochondrial apoptosis-inducing factor[J]. Nature, 1999, 397(6718): 441-446.
[20] Zamzami N, Marchetti P, Castedo M, et al. Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo[J]. J Exp Med, 1995, 181(5): 1661-1672.
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