连翘苷调控细胞因子信号转导抑制因子1对肺炎链球菌诱导的肺泡上皮细胞凋亡和氧化应激的影响

Effects of phillyrin on alveolar epithelial cell apoptosis and oxidative stress induced by Streptococcus pneumoniae by regulating suppressor of cytokine signaling 1

目的研究连翘苷调控细胞因子信号转导抑制因子1(SOCS1)对肺炎链球菌诱导的肺泡上皮细胞凋亡和氧化应激影响。方法将人Ⅱ型肺泡上皮细胞分为对照组、模型组(肺炎链球菌感染)、实验低、中、高剂量组(在模型组基础上分别给予5、10、20μmol·L^(-1)连翘苷)、实验高剂量+si-NC组(肺炎链球菌感染,转染siRNA NC、20μmol·L^(-1)连翘苷处理)、实验高剂量+si-SOCS1组(肺炎链球菌感染,转染siRNA SOCS1、20μmol·L^(-1)连翘苷处理)。用蛋白质印迹(Western blot)法检测SOCS1蛋白表达;用细胞计数试剂盒-8(CCK-8)检测细胞增殖;用碘化丙啶(PI)单染法检测细胞周期;用Annexin V-PI双染法检测细胞凋亡;用可见分光光度法检测丙二醛(MDA);用氮蓝四唑(NBT)法检测超氧化物歧化酶(SOD)。结果对照组、模型组、实验高剂量组、实验高剂量+si-NC组和实验高剂量+si-SOCS1组肺泡上皮细胞SOCS1蛋白表达水平分别为1、0.54±0.04、0.96±0.04、0.88±0.03和0.40±0.07;G0/G1期细胞比例分别为(42.04±7.08)%、(69.75±3.31)%、(44.84±2.05)%、(43.72±3.09)%和(61.26±6.17)%;凋亡率分别为(6.38±0.47)%、(30.05±2.06)%、(12.20±1.25)%、(12.44±2.36)%和(17.84±1.48)%;SOD含量分别为(132.30±9.79)、(59.04±5.80)、(113.70±10.38)、(111.38±8.34)和(71.89±9.01)U·mg^(-1),以上指标,模型组与对照组、实验高剂量组比较,实验高剂量+si-SOCS1组与实验高剂量+si-NC组比较,差异均有统计学意义(均P<0.05)。结论连翘苷促进SOCS1表达减少肺炎链球菌诱导的肺泡上皮细胞凋亡和氧化应激。

Objective To investigate the effect of phillyrin regulation of suppressor of cytokine signaling 1(SOCS1)on apoptosis and oxidative stress induced by Streptococcus pneumoniae in alveolar epithelial cells.Methods Human type II alveolar epithelial cells were divided into control group,model group(Streptococcus pneumoniae infection),experimental-L,-M and-H groups(5,10,20μmol·L^(-1) phillyrin were added on the basis of model group),experimental-H+si-NC group(Streptococcus pneumoniae infection,transfected with siRNA NC,20μmol·L^(-1) phillyrin treatment),experimental-H+si-SOCS1 group(Streptococcus pneumoniae infection,transfected with siRNA SO CS 1,20μmol·L^(-1) phillyrin treatment).Western blot detects SOCS1 protein expression;cell counting kit-8(CCK-8)detects cell proliferation;propidium iodide(PI)monochromaticity detects cell cycle;annexin V-PI double staining detects apoptosis;visible spectrophotometry detects malondialdehyde(MDA),and nitrogen blue tetrazole(NBT)detects superoxide dismutase(SOD).Results The expression levels of SOCS1 protein in alveolar epithelial cells of control group,model group,experimental-H group,experimental-H+si-NC group and experimental-H+si-SOCS1 group were 1,0.54±0.04,0.96±0.04,0.88±0.03 and 0.40±0.07,respectively.The percentage of G0/G1 phase cells was(42.04±7.08)%,(69.75±3.31)%,(44.84±2.05)%,(43.72±3.09)%and(61.26±6.17)%,respectively.The apoptosis rates were(6.38±0.47)%,(30.05±2.06)%,(12.20±1.25)%,(12.44±2.36)%and(17.84±1.48)%,respectively.The contents of SOD were(132.30±9.79),(59.04±5.80),(113.70±10.38),(111.38±8.34)and(71.89±9.01)U·mg^(-1),respectively.There were significant differences in the above indexes between the model group and the control group,and between the experimental-H+si-SOCS1 group and the experimental-H+si-NC group(all P<0.05).Conclusion Phillyrin promotes SOCS1 expression and reduces apoptosis and oxidative stress induced by Streptococcus pneumoniae in alveolar epithelial cells.