罗红霉素对离体哮喘大鼠气道平滑肌细胞凋亡的影响及其机制  

The effect and the mechanism of Roxithromycin on apoptosis of airway smooth muscle cells from asthmatic rats in vitro

作　　者：吴海亚[1] 戴元荣[2] 应斌宇[1] 

机构地区：[1]温州医科大学附属第二医院重症医学科,浙江温州325027 [2]温州医科大学附属第二医院呼吸内科,浙江温州325027

出　　处：《温州医学院学报》2015年第4期252-255,259,共5页Journal of Wenzhou Medical College

基　　金：浙江省自然科学基金资助项目(Y2080466);浙江省医药卫生科技计划项目(2009A144)

摘　　要：目的:观察罗红霉素对离体气道平滑肌细胞(ASMCs)凋亡的影响以及线粒体凋亡途径在ASMCs凋亡中的作用。方法:体外培养哮喘大鼠ASMCs,用不同浓度(0、10、25、50、100μg/m L分别对应R0组、R10组、R25组、R50组、R100组)的罗红霉素干预48 h。Annexin V/PI双标记流式细胞术检测早期细胞凋亡;JC-1染色流式细胞术分析线粒体跨膜电位(ΔΨm)的变化;Western blot法分析线粒体内和胞浆内细胞色素C(Cyt-c)的含量。结果:各实验组(R10组、R25组、R50组、R100组)ASMCs的早期凋亡率分别为(4.6±1.9)%、(5.8±2.9)%、(12.0±5.6)%、(26.9±11.1)%,较R0组的(2.9±1.7)%高,其中R100组与R0组间差异有统计学意义(x2=13.30,P<0.01)。各实验组(R10组、R25组、R50组、R100组)低ΔΨm细胞所占比例分别为(27.88±13.10)%、(40.35±9.19)%、(48.40±14.15)%、(52.90±15.88)%,较R0组的(19.78±9.85)%高,其中R100组和R50组ΔΨm显著低于R0组和R10组(P<0.05或0.01),R25组与R0组比较,差异有统计学意义(P<0.05),R10组与R0组比较差异无统计学意义(P>0.05);各实验组(R10组、R25组、R50组、R100组)胞浆Cyt-c的含量分别为(0.87±0.19)、(0.97±0.17)、(1.01±0.12)、(1.14±0.07),较R0组的(0.67±0.12)高,R100组与R0组比较差异有统计学意义(x2=9.075,P<0.05)。结论:罗红霉素可以通过激活线粒体凋亡通路促进离体ASMCs的凋亡。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 asth-matic rats were cultured in vitro and intervened with various concentrations of Roxithromycin for 48 hours. Early apoptotic cells were detected through lfow cytometry （FCM） after stained with Annexin V/PI;Alteration of mito-chondrial 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 signiifcant difference （x^2=13.30, P〈0.01） between the group R100 and the control. RXM induced loss ofΔΨm：The proportion of lowΔΨm 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 con-trol group （19.78±9.85）%. The indexes of group R100 and R50 were signiifcantly lower than that of group R0 and R10 （P〈0.05 or 0.01）. The difference between group R25 and R0 was also statistically signiifcance （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 respec-tively （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 signiifcant difference between the group R100 and R0 （x^2=9.075, P〈0.05）. Conclusion：Roxithromycin can induce apoptosis of ASMCs in vitro via activating mitochondrial pathway.

关 键 词：罗红霉素 哮喘 气道重塑 平滑肌细胞 凋亡 

分 类 号：R562.2[医药卫生—呼吸系统]