短链脂肪酸对胰腺星状细胞氧化应激和活化的调控作用  

Regulatory effects of short-chain fatty acids on oxidative stress and activation of pancreatic stellate cells

作　　者：陆宏娜 许丰 张秋波 翁烃 张良舜 李先鹏 Lu Hongna;Xu Feng;Zhang Qiubo;Weng Ting;Zhang Liangshun;Li Xianpeng(Department of Gastroenterology,The Affiliated Lihuili Hospital of Ningbo University,Ningbo 315103,China;Abdominal Transplantation Center,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]宁波大学附属李惠利医院消化内科,宁波315103 [2]上海交通大学附属瑞金医院腹部器官移植中心,上海200025

出　　处：《中华胰腺病杂志》2024年第3期210-215,共6页Chinese Journal of Pancreatology

基　　金：宁波市自然科学基金(2016A610189)。

摘　　要：目的探讨短链脂肪酸(SCFA)对缺氧诱导的胰腺星状细胞(PSCs)氧化应激和活化的调控作用。方法常氧及缺氧培养PSCs建立常氧组和缺氧组,SCFA工作液(10 mmol/L乙酸钠、0.5 mmol/L丙酸钠及0.5 mmol/L丁酸钠)和生理盐水分别预处理PSCs后进行缺氧培养建立缺氧SCFA组和缺氧对照组。采用CCK-8法检测各组PSCs生长活力,采用DCFH-DA荧光探针法检测PSCs活性氧(ROS)的相对水平,JC-1荧光探针检测线粒体膜电位的变化,蛋白质免疫印迹法检测PSCs周期相关蛋白cyclin A和cyclin D、缺氧标志蛋白HIF1α、活化标志蛋白α-SMA及抗氧化蛋白NRF2和HO-1的表达。结果缺氧组PSCs在培养48 h时的相对活力显著高于常氧组(1.23±0.05比0.99±0.04),而缺氧SCFA组在培养36 h和48 h时的相对活力均显著低于缺氧对照组(0.69±0.01比0.86±0.03,0.86±0.02比1.25±0.05)。缺氧组ROS相对水平显著高于常氧组(1.74±0.11比1.00±0.10),而缺氧SCFA组ROS相对水平显著低于缺氧对照组(1.39±0.14比1.66±0.11)。缺氧组JC-1聚合体的荧光信号明显高于常氧组(1.36±0.05比1.00±0.11),而缺氧SCFA组JC-1聚合体的荧光信号明显低于缺氧对照组(1.11±0.03比1.32±0.06)。缺氧组cyclin A、cyclin D、HIF1α、α-SMA、NRF2和HO-1表达量显著高于常氧组(1.19±0.01比0.63±0.02,0.93±0.02比0.83±0.03,1.18±0.07比0.41±0.02,1.19±0.14比0.66±0.04,1.22±0.11比0.61±0.04,1.28±0.12比0.68±0.02),而缺氧SCFA组cyclin A、cyclin D、α-SMA、NRF2和HO-1的表达量显著低于缺氧对照组(0.79±0.04比1.15±0.03,0.88±0.01比0.95±0.03,0.87±0.01比1.18±0.05,0.84±0.01比1.22±0.04,0.92±0.02比1.27±0.06)。以上差异均有统计学意义(P值均<0.05)。结论SCFA显著改善缺氧状态下PSCs的氧化应激状态,维持线粒体膜电位的稳定,抑制缺氧诱导的PSCs活化。Objective To explore regulatory effects of short-chain fatty acids(SCFA)on hypoxia-induced oxidative stress and activation of pancreatic stellate cells(PSCs).Methods PSCs were cultured in normoxia or hypoxia conditions to establish normoxia or hypoxia group.PSCs were pre-treated with SCFA working solution(10 mmol/L sodium acetate,0.5 mmol/L sodium propionate and 0.5 mmol/L sodium butyrate),and then cultured in hypoxia conditions to establish the hypoxia-SCFA group.PSCs pre-treated by normal saline was set as the hypoxia-control group.The relative growth viability of the cells was detected by the CCK-8 assay.Relative levels of reactive oxygen species(ROS)were detected by DCFH-DA fluorescence probe method.The mitochondrial membrane potential was detected by JC-1 fluorescence probe.Protein expression of cyclin-associated marker cyclin A and cyclin D,hypoxic marker HIF1α,activation markerα-SMA,and antioxidant marker NRF2 and HO-1 was detected by western blotting.Results The relative viability of PSCs in hypoxia group was significantly higher than that in normoxia group at 48 h(1.23±0.05 vs 0.99±0.04),but the relative viability of hypoxia-SCFA group was significantly lower than that of the hypoxic-control group at both 36 h and 48 h(0.69±0.01 vs 0.86±0.03,0.86±0.02 vs 1.25±0.05).The relative level of ROS was significantly higher in hypoxia group than normoxia group(1.74±0.11 vs 1.00±0.10).The relative level of ROS was significantly lower in the hypoxia-SCFA group than the hypoxia-control group(1.39±0.14 vs 1.66±0.11).The fluorescence signals of JC-1 polymer in hypoxia group were significantly higher than those in normoxia group(1.36±0.05 vs 1.00±0.11),whereas the fluorescence signals of JC-1 polymer were significantly lower in hypoxia-SCFA group than in hypoxia-control group(1.11±0.03 vs 1.32±0.06).The expression of cyclin A,cyclin D,HIF1α,α-SMA,NRF2,and HO-1 was significantly higher in hypoxia group than those in normoxia group(1.19±0.01 vs 0.63±0.02,0.93±0.02 vs 0.83±0.03,1.18±0.07 vs 0.41±0.

关 键 词：脂肪酸类 挥发性 胰腺星形细胞 低氧 氧化性应激 活化 

分 类 号：R576[医药卫生—消化系统]