摘要
目的:研究大黄素对急性胰腺炎胰腺腺泡细胞外泌体水平和氧化应激的影响。方法:磁珠结合流式细胞仪检测35例正常健康人和35例急性胰腺炎患者血清中外泌体表面抗原CD69的表达水平。用10-7 mol/L雨蛙素诱导大鼠胰腺外分泌细胞AR42J 24 h建立炎性损伤模型,设不处理的细胞作为对照,用ELISA法检测细胞培养上清中淀粉酶(AMY)活性、TNF-α含量。用大黄素(0、10、20、30μmol/L)处理AR42J模型细胞48 h,MTT实验检测细胞增殖抑制率;筛选最适浓度20μmol/L大黄素处理的模型细胞为大黄素组,DMSO处理的模型细胞作为阴性对照组,另设模型组,用ELISA法检测培养上清中SOD、GSH、MDA活性,磁珠结合流式细胞仪检测细胞CD69的表达。结果:与正常对照组比较,急性胰腺炎组患者血清CD69表达升高(P<0.001)。与对照组相比,模型组细胞培养上清中AMY活性、TNF-α含量均升高(P<0.05)。与其他2组相比,大黄素组细胞培养上清中SOD、GSH活性升高,MDA活性、CD69表达水平降低(P<0.05)。结论:大黄素可抑制急性胰腺炎胰腺腺泡细胞的氧化应激反应,下调外泌体水平。
Aim:To study the effects of emodin on exosome level and oxidative stress of pancreatic acinar cells in acute pancreatitis.Methods:Magnetic bead combined with flow cytometry was used to detect the expression of exosome surface antigen CD69 in the serum of 35 healthy people and 35 patients with acute pancreatitis.The inflammatory damage model of rat pancreatic exocrine cells AR42J was induced with 10-7 mol/L cerulein for 24 hours,and the normal cells were used as control.The activity of amylase(AMY)and TNF-αcontent in the cell supernatant were measured by ELISA.Emodin(0,10,20,30μmol/L)were used to treat AR42J model cells for 48 hours.MTT experiment was used to detect the cell proliferation inhibition rate and 20μmol/L was selected as the optimal concentration of emodin.The model cells were allocated into emodin group(treated by 20μmol/L emodin),negative control group(treated by DMSO),and the model group.The activities of SOD,GSH,and MDA were detected by ELISA,and the expression of CD69 in the cells was detected by magnetic beads combined with flow cytometry.Results:Compared with the control group,the expression of CD69 significantly increased in patients with acute pancreatitis(P<0.001).Compared with the control group,the activity of AMY and the content of TNF-αin the model group significantly increased(P<0.05).Compared with the other 2 groups,the activity of SOD and GSH in the emodin group significantly increased,and the activity of MDA and the expression of CD69 significantly reduced(P<0.05).Conclusions:Emodin could inhibit the oxidative stress and down-regulate exosome level of pancreatic acinar cells in acute pancreatitis.
作者
蔡青云
许丽君
CAI Qingyun;XU Lijun(Department of Emergency, Zhumadian Central Hospital, Zhumadian, Henan 463000;Department of Emergency, Henan Provincial People′s Hospital, Zhengzhou 450003)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2020年第3期373-376,共4页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省科技攻关项目(124105410258)。
