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 cas-pase-3、RhoA、ROCK表达情况.结果显示,与Control组相比,Model组细胞的D490、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D490、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D490、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.
Abstract
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 Con-trol 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 estab-lished 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(malo-ndialdehyde)were detected by kits.The levels of IL-10(interleukin-10),IL-1 β and 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 D490,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.Com-pared with the Model group,the D490,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 D490,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 apoptosis,and al-leviating cell injury.
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