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
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-.
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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