益母草碱调节MCP-1/CCR2信号轴对LPS诱导的肺泡上皮细胞炎性损伤的影响  

Effects of leonzurine on LPS-induced inflammatory injury of alveolar epithelial cells by regulating the MCP-1/CCR2 axis

作　　者：蔡文宇 张米斯 杨雪婷 CAI Wenyu;ZHANG Misi;YANG Xueting(Department of Emergency,the Fifth People’s Hospital of Sichuan,Sichuan,Chengdu 610036,China;不详)

机构地区：[1]四川省第五人民医院急诊科,成都市610036 [2]四川省成都市第三人民医院蒲江医院呼吸内科 [3]四川省人民医院急诊科

出　　处：《河北医药》2024年第7期983-987,共5页Hebei Medical Journal

基　　金：四川省卫生健康委员会科研课题(编号:19ZD002)。

摘　　要：目的探讨益母草碱(Leo)调节单核细胞趋化蛋白-1(MCP-1)/趋化因子受体-2(CCR2)信号轴对脂多糖(LPS)诱导的肺泡上皮细胞炎性损伤的影响。方法体外培养人肺泡上皮细胞(AEC);将细胞分为对照组、模型组(LPS组,10μg/mL LPS)、低浓度Leo组(Leo-L组,10μmol/L Leo)、中浓度Leo组(Leo-M组,20μmol/L Leo)、高浓度Leo组(Leo-H组,30μmol/L Leo)、MCP-1激活剂SLIGKV组(SLIGKV组,50μmol/L SLIGKV)、高浓度Leo+MCP-1激活剂SLIGKV组(Leo-H+SLIGKV组,30μmol/L Leo-H+50μmol/L SLIGKV);CCK-8实验检测细胞增殖;流式细胞术检测细胞凋亡;酶联免疫吸附测定(ELISA)法检测细胞上清中白细胞介素-2(IL-2)、肿瘤坏死因子α(TNF-α)水平;实时荧光定量PCR(qRT-PCR)和Western blot分别检测细胞中MCP-1/CCR2信号通路及凋亡相关因子表达水平。结果与对照组比较,LPS组AEC细胞凋亡率、MCP-1、CCR2、IL-2、TNF-α及Bcl-2相关X蛋白(Bax)表达显著升高,OD 450值和B淋巴细胞瘤-2(Bcl-2)表达显著降低(P<0.05);与LPS组比较,Leo-L组、Leo-M组、Leo-H组AEC细胞凋亡率、MCP-1、CCR2、IL-2、TNF-α及Bax表达显著降低,OD 450值和Bcl-2表达显著升高,且呈现剂量依赖性(P<0.05);SLIGKV组上述指标趋势相反;SLIGKV减弱了高剂量Leo对LPS诱导的肺泡上皮细胞炎性损伤的改善作用。结论Leo可能通过抑制MCP-1/CCR2信号轴改善LPS诱导的肺泡上皮细胞炎性损伤。Objective To investigate the effect of leonzurine(Leo)on lipopolysaccharide(LPS)-induced inflammatory injury of alveolar epithelial cells by regulating the monocyte chemoattractant protein 1(MCP-1)/CC chemokine receptor 2(CCR2)axis.Methods The human alveolar epithelial cells(AECs)were cultured in vitro.They were induced with blank control,10μg/mL LPS,LPS+low-dose Leo at 10μmol/L,LPS+medium-dose Leo at 20μmol/L,LPS+high-dose Leo at 30μmol/L,LPS+MCP-1 activator SLIGKV at 50μmol/L,and LPS+high-dose Leo+SLIGKV.Cell proliferation and apoptosis were detected by CCK-8 assay and flow cytometry,respectively.Relative levels of interleukin 2(IL-2)and tumor necrosis factor-α(TNF-α)in the cell supernatant were measured by enzyme linked immunosorbent assay(ELISA).Real-time fluorescence quantitative PCR(qRT-PCR)and Western blot were applied to detect the expression levels of key genes in the MCP-1/CCR2 signaling pathway and apoptotic proteins.Results Compared with those of blank control,LPS-induced AECs presented significantly higher apoptotic rate and expression levels of MCP-1,CCR2,IL-2,TNF-α and Bcl-2-associated X protein(Bax),and significantly lower optical density at 450 nm(OD450)and expression level of B-cell lymphoma-2(Bcl-2)(P<0.05).Compared with those induced with LPS,LPS-induced AECs treated with low-dose,medium-dose and high-dose Leo presented significantly lower apoptotic rate and expression levels of MCP-1,CCR2,IL-2,TNF-α and Bax,and significantly higher OD450 and expression level of Bcl-2 in a dose-dependent manner(P<0.05).Opposite trends were detected in LPS-induced cells treated with SLIGKV,which weakened the protective effect of high-dose Leo on LPS-induced inflammatory injury of AECs.Conclusion Leo alleviates LPS-induced inflammatory damage to alveolar epithelial cells by inhibiting the MCP-1/CCR2 axis.

关 键 词：益母草碱 脂多糖 单核细胞趋化蛋白1/趋化因子受体-2信号轴 肺泡上皮细胞 增殖 凋亡 炎症 

分 类 号：R563[医药卫生—呼吸系统]