南蛇藤素调节SDF-1/CXCR4信号通路对脂多糖诱导的A549细胞损伤的影响  

Influence of Celastrol on LPS-inducedA549 cells injury by regulating SDF-1/CXCR4 signaling pathway

作　　者：符丽娜 邢博真 陈瑶 潘相静 符丽云 孙少清 Fu Lina;Xing Bozhen;Chen Yao;Pan Xiangjing;Fu Liyun;Sun Shaoqing(Department of Respiratory,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China;Department of Infection,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China;Department of Radiology,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南省海口市海南医学院第一附属医院呼吸科,海口570102 [2]海南省海口市海南医学院第一附属医院感染科,海口570102 [3]海南省海口市海南医学院第一附属医院放射科,海口570102

出　　处：《中华细胞与干细胞杂志（电子版）》2023年第3期144-150,共7页Chinese Journal of Cell and Stem Cell（Electronic Edition）

摘　　要：目的分析南蛇藤素对脂多糖(LPS)诱导的肺泡上皮细胞损伤的影响以及基质细胞衍生因子-1(SDF-1)/趋化因子受体4(CXCR4)通路在其中发挥的作用。方法培养人肺泡Ⅱ型上皮细胞系A549细胞,用浓度为10μg/mL LPS处理24 h,并在LPS处理前用终浓度为25、100 mmol/L的南蛇藤素或100 mmol/L的南蛇藤素与100μg/mL的AMD3100(CXCR4抑制剂)处理2 h。CCK-8法测定A549细胞活力,TUNEL法测定A549细胞凋亡,试剂盒法测定A549细胞培养上清液中乳酸脱氢酶(LDH)含量,ELISA法检测A549细胞肿瘤坏死因子(TNF)-α、白细胞介素6(IL-6)、IL-10水平,Western blot法测定SDF-1、CXCR4蛋白表达。多组间比较采用单因素方差分析,进一步两两比较采用SNK-q法进行。结果与对照组相比,LPS组A549细胞TUNEL阳性率[(65.78±8.21)%比(4.23±0.77)%]、细胞凋亡率[(21.38±2.70)%比(7.64±0.95)%]、cleaved caspase-3、Bax蛋白、LDH、TNF-α、IL-6、SDF-1(2.02±0.25比0.75±0.10)、CXCR4蛋白(1.59±0.19比0.71±0.08)升高(P<0.05),存活率、Bcl-2蛋白、IL-10表达降低(P<0.05);与LPS组相比,LPS+南蛇藤素低剂量组、LPS+南蛇藤素高剂量组A549细胞TUNEL阳性率[(52.51±6.54)%,(41.37±5.17)%比(65.78±8.21)%]、细胞凋亡率[(17.62±2.45)%,(12.37±1.54)%比(21.38±2.70)%]、cleaved caspase-3、Bax蛋白、LDH、TNF-α、IL-6、SDF-1(1.69±0.20,1.32±0.18比2.02±0.25)、CXCR4蛋白(1.05±0.13,1.31±0.16比1.59±0.19)降低,存活率、Bcl-2蛋白和IL-10表达升高(P<0.05);与LPS+南蛇藤素高剂量组相比,LPS+南蛇藤素高剂量+AMD3100组A549细胞TUNEL阳性率[(24.28±4.28)%比(41.37±5.17)%]、细胞凋亡率[(9.91±1.25)%比(12.37±1.54)%]、cleaved caspase-3、Bax蛋白、LDH、TNF-α、IL-6、SDF-1(0.95±0.13比1.32±0.18)、CXCR4蛋白(0.78±0.09比1.05±0.13)降低,存活率、Bcl-2蛋白和IL-10表达升高(P<0.05)。结论南蛇藤素可降低LPS诱导的A549细胞凋亡与炎症,提高细胞存活,其作用机制可能与抑制SDF-1/CXCR4通路有关。Objective To analyze the influence of Celastrol on lipopolysaccharide(LPS)-induced alveolar epithelial cell injury and the role of stromal cell-derived factor-1(SDF-1)/chemokine receptor 4(CXCR4)pathway in it.Methods Human alveolar typeⅡepithelial cell line A549 cells were cultured and treated with 10μg/mL LPS for 24 h.Before LPS treatment,treat them with final concentrations of 25 mmol/L,100 mmol/L of celastrol or 100 mmol/L of celastrol and 100μg/L of AMD3100(CXCR4 inhibitor)for 2 hours.The viability of A549 cells was determined by the CCK-8 method,apoptosis of A549 cells was measured by the TUNEL method,the LDH of A549 cells was determined by kits,the levels of tumor necrosis factor(TNF)-α,interleukin-6(IL-6)and IL-10 in A549 cells were detected by ELISA,and the expression of SDF-1 and CXCR4 protein were as detected by Western blot.One-way analysis of variance was used for statistical comparison among multiple groups,and the SNK-q method was used for further pairwise statistical comparison.Results Compared with the control group,the positive rate of TUNEL[(65.78±8.21)%vs(4.23±0.77)%],the apoptosis rate[(21.38±2.70)%vs(7.64±0.95)%],cleaved caspase-3,Bax protein,LDH,TNF-α,IL-6,SDF-1(2.02±0.25 vs 0.75±0.10),and CXCR4(1.59±0.19 vs 0.71±0.08)protein were increased in A549 cells in LPS group(P<0.05),and the survival rate,Bcl-2 protein and IL-10 expression decreased(P<0.05);compared with the LPS group,the positive rate of TUNEL[(52.51±6.54)%,(41.37±5.17)%vs(65.78±8.21)%],the apoptosis rate was[(17.62±2.45)%,(12.37±1.54)%vs(21.38±2.70)%],cleaved caspase-3,Bax protein,LDH,TNF-α,IL-6,SDF-1(1.69±0.20,1.32±0.18 vs 2.02±0.25),CXCR4(1.31±0.16,1.05±0.13 vs 1.59±0.19)protein in A549 cells in the low-dose Celastrol group and high-dose Celastrol group significantly reduced,and the survival rate,Bcl-2 protein and IL-10 expression significantly increased(P<0.05);compared with the high-dose Celastrol group,the positive rate of TUNEL[(24.28±4.28)%vs(41.37±5.17)%],the apoptosis rate[(9.91±1.25)%vs(12.37

关 键 词：南蛇藤素 SDF-1 CXCR4 A549细胞 

分 类 号：R73[医药卫生—肿瘤]