中国临床药理学杂志2025，Vol.41Issue(23)：3358-3365,8.DOI:10.13699/j.cnki.1001-6821.2025.23.009
SESN2激活ULK1介导JNK信号通路在肝星状细胞活化中的调节机制
SESN2 activation of ULK1 mediates the regulatory mechanism of JNK signaling pathway in hepatic stellate cell activation
摘要
Abstract
Objective To investigate the effect of sestrin 2(SESN2)on JS-1 cell activation through unc-51-like kinase 1(ULK1)and its mechanism.Methods The JS-1 cells were divided into seven groups:the control group(cultured under normal conditions),si-NC group(transfected with si-NC),the si-SESN2 group(transfected with si-SESN2),model group(cultured in medium containing 5 μg·L-1 transforming growth factor-β1 for 24 h),model+oe-SESN2 group(transfected with oe-SESN2 on the basis of the model group),model+si-SESN2 group(transfected with si-SESN2 on the basis of the model group),and model+si-SESN2+oe-ULK1 group(co-transfected with si-SESN2 and oe-ULK1 on the basis of the model group).The levels of fibrous actin(F-actin)was detected by immunofluorescence.Quantitative real time polymerase chain reaction was used to detect the ULK1 mRNA level.The relevtive expression levels of SESN2,light chain 3 Ⅱ/Ⅰ (LC3 Ⅱ/Ⅰ),connective tissue growth factor(CTGF),collagen type Ⅰ α1(Col1 a1),phospho-c-Jun N-terminal kinase(p-JNK),c-Jun N-terminal kinase(JNK),c-Jun and activating transcription factor 2(ATF2)were detected by Western Blot.Cell proliferation rate was detected by cell counting kit-8.Transwell assesses the ability of cells to invade.Results The relative expression levels of SESN2 protein in the control group,model group,model+oe-SESN2 group,and model+si-SESN2 group were 1.00±0.19,2.57±0.49,3.61±0.68,and 1.66±0.31,respectively;the relative fluorescence intensity of F-actin was 1.00±0.18,4.57±0.92,6.33±1.19,and 2.81±0.57,respectively;the relative expression levels of ULK1 mRNA were 1.00±0.15,2.67±0.48,3.52±0.66,and 1.95±0.37,respectively.The relative expression levels of LC3 Ⅱ/Ⅰ protein in the control group,model group,model+oe-SESN2 group,model+si-SESN2 group,and model+si-SESN2+oe-ULK1 group were 1.00±0.20,2.79±0.53,3.57±0.68,1.88±0.35,and 2.51±0.48,respectively;the cell proliferation rates were(100.00±2.23)％,(141.31±4.17)％,(156.52±8.84)％,(109.49±7.35)％,and(127.08±8.16)％,respectively;the number of cell invasions was 66.04±11.45,137.84±21.73,215.19±35.28,75.46±13.51,and 124.27±19.50,respectively;the relative expression levels of CTGF protein were 1.00±0.17,2.65±0.51,3.77±0.72,1.51±0.29,and 2.53±0.51,respectively;the relative expression levels of Col1a1 protein were 1.00±0.15,2.93±0.57,3.88±0.75,1.61±0.31,and 2.72±0.52,respectively;the relative phosphorylation levels of JNK protein were 1.00±0.16,2.21±0.42,3.08±0.59,1.46±0.29,and 2.04±0.37,respectively.Compared with the control group,the above indicators of the model group showed statistically significant differences;compared with the model group,the above indicators of the model+oe-SESN2 group or model+si-SESN2 group showed statistically significant differences;compared with the model+si-SESN2 group,the above indicators of the model+si-SESN2+oe-ULK1 group all showed statistically significant differences(P＜0.05,P＜0.01,P＜0.001).Conclusion SESN2 activates ULK1,promotes JS-1 cell activation,promotes cell proliferation and invasion,and regulates cell homeostasis,possibly through the activation of JNK signaling pathway.关键词
应激诱导蛋白2/unc-51样激酶1/肝星状细胞/应激活化蛋白激酶信号通路/细胞增殖Key words
sestrin 2/unc-51-like kinase 1/hepatic stellate cells/c-Jun N-terminal kinase signaling pathway/cell proliferation分类
医药卫生引用本文复制引用
汤洁,马天奇,姜正伟,景源伟,孙路翔,程宏..SESN2激活ULK1介导JNK信号通路在肝星状细胞活化中的调节机制[J].中国临床药理学杂志,2025,41(23):3358-3365,8.基金项目
2024年度淮安市卫生健康科研基金资助项目(HAWJZ2024003) （HAWJZ2024003）
