摘要
该文探讨了橄榄苦苷(OP)调节Ras同源基因家族成员A(RhoA)/Rho相关卷曲螺旋形成蛋白激酶(ROCK)信号通路对肺炎链球菌感染导致的肺泡上皮细胞损伤的影响。将肺泡上皮细胞A549随机分为Control组、Model组、L-OP组、M-OP组、H-OP组、U46619组(H-OP+RhoA/ROCK信号通路激活剂U46619),除Control组正常培养外,其余各组均以肺炎链球菌感染建立细胞损伤模型。MTT和平板克隆实验检测细胞增殖情况;试剂盒检测细胞中超氧化物歧化酶(SOD)、活性氧(ROS)、丙二醛(MDA)水平;ELISA检测细胞中白细胞介素10(IL-10)、IL-1β、肿瘤坏死因子-α(TNF-α)水平;流式细胞仪检测细胞凋亡情况;Western blot检测细胞中Bcl-2、Bax、cleaved caspase-3、RhoA、ROCK表达情况。结果显示,与Control组相比,Model组细胞的D_(490)、克隆数、SOD、IL-10、Bcl-2表达水平降低,细胞凋亡率、ROS、MDA、IL-1β、TNF-α、Bax、cleaved caspase-3、RhoA、ROCK表达水平升高;与Model组相比,L-OP组、M-OP组、H-OP组细胞的D_(490)、克隆数、SOD、IL-10、Bcl-2表达水平升高,细胞凋亡率、ROS、MDA、IL-1β、TNF-α、Bax、cleaved caspase-3、RhoA、ROCK表达水平降低;与H-OP组相比,U46619组细胞的D_(490)、克隆数、SOD、IL-10、Bcl-2表达水平降低,细胞凋亡率、ROS、MDA、IL-1β、TNF-α、Bax、cleaved caspase-3、RhoA、ROCK表达水平升高。由此提示,OP可能是通过抑制RhoA/ROCK信号通路,减轻肺炎链球菌感染导致的肺泡上皮细胞炎症与氧化应激反应,进而促进细胞增殖,抑制细胞凋亡,缓解细胞损伤的。
The aim of this study is to investigate the effect of OP(oleuropein)on alveolar epithelial cell injury induced by Streptococcus pneumoniae infection by regulating RhoA(Ras homologous gene family member A)/ROCK(Rho-associated coiled-coil kinase)signaling pathway.Alveolar epithelial cells A549 were randomly divided into Control group,Model group,L-OP group,M-OP group,H-OP group and U46619 group(H-OP+RhoA/ROCK signaling pathway activator U46619).Except for the normal culture of the Control group,the cell injury model was established with Streptococcus pneumoniae infection in the other groups.MTT and plate cloning were used to detect the proliferation of cells.The levels of SOD(superoxide dismutase),ROS(reactive oxygen species)and MDA(malondialdehyde)were detected by kits.The levels of IL-10(interleukin-10),IL-1βand TNF-α(tumor necrosis factor-α)were detected by ELISA.Flow cytometry was used to detect apoptosis.Western blot was used to detect the expression of Bcl-2,Bax,cleaved caspase-3,RhoA and ROCK.The result showed that compared with the Control group,the D_(490),clone number,the expression levels of SOD,IL-10 and Bcl-2 in the Model group were decreased;the apoptosis rate,the expression levels of ROS,MDA,IL-1β,TNF-α,Bax,cleaved caspase-3,RhoA and ROCK were increased.Compared with the Model group,the D_(490),clone number,the expression levels of SOD,IL-10 and Bcl-2 in L-OP group,M-OP group and H-OP group were increased;the apoptosis rate,the expression levels of ROS,MDA,IL-1β,TNF-α,Bax,cleaved caspase-3,RhoA and ROCK were decreased.Compared with the H-OP group,the D_(490),clone number,the expression levels of SOD,IL-10 and Bcl-2 in the U46619 group were decreased;the apoptosis rate,the expression levels of ROS,MDA,IL-1β,TNF-α,Bax,cleaved caspase-3,RhoA and ROCK were increased.As a reminder,OP may alleviate inflammation and oxidative stress in alveolar epithelial cells infected with Streptococcus pneumoniae by inhibiting the RhoA/ROCK signaling pathway,thereby promoting cell proliferation,inhibiting cell a
作者
严娅芬
陆光兵
田良东
张东香
YAN Yafen;LU Guangbing;TIAN Liangdong;ZHANG Dongxiang(Department of Respiratory and Critical Care Medicine,Meishan Traditional Chinese Medicine Hospital,Meishan 620010,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2024年第9期1663-1671,共9页
Chinese Journal of Cell Biology
