茯苓酸通过抑制cGAS-STING信号通路减轻溃疡性结肠炎大鼠结肠上皮细胞损伤  

Pachymic acid alleviates colonic epithelial cell damage in ulcerative colitis rats by inhibiting cGAS-STING signaling pathway

作　　者：张士伟 程慎令[1] 邢启峰 ZHANG Shiwei;CHENG Shenling;XING Qifeng(Department of Pharmacy,Tengzhou Central People's Hospital,Tengzhou 277599,China)

机构地区：[1]滕州市中心人民医院药学部,277599

出　　处：《免疫学杂志》2023年第8期672-680,共9页Immunological Journal

基　　金：山东省中医药科技发展计划项目(2019-0643)。

摘　　要：目的探讨茯苓酸(PA)调节环磷酸鸟苷-腺苷酸合成酶(cGAS)-干扰素刺激基因(STING)信号通路对溃疡性结肠炎大鼠肠上皮细胞损伤的影响。方法将SD大鼠分为对照组、溃疡性结肠炎组、大鼠PA低剂量组(R-PA-L组)、大鼠PA中剂量组(R-PA-M组)、大鼠PA高剂量组(R-PA-H组),大鼠RocA(cGAS-STING通路激活剂)组(R-RocA组)、R-PA-H+RocA组;统计大鼠体质量下降及疾病活动指数(DAI)评分;HE染色检测大鼠结肠组织病理变化。分离并鉴定对照组、溃疡性结肠炎组大鼠肠上皮细胞,并分组为NC组、Model组、PA低剂量组(PA-L组,2μmol/L)、PA中剂量组(PA-M组,4μmol/L)、PA高剂量组(PA-H组,8μmol/L)、RocA组(25 nmol/L)、PA-H+RocA组(8μmol/L+25 nmol/L);ELISA法检测细胞上清中白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)、IL-6水平;试剂盒检测大鼠结肠上皮细胞中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;流式细胞术检测大鼠结肠上皮细胞凋亡;Western blot检测大鼠结肠上皮细胞中Bcl-2相关X蛋白(Bax)、多聚腺苷二磷酸核糖聚合酶(PARP)、cGAS、p-STING、TANK结合酶1(TBK1)、干扰素调节因子-3(IRF-3)蛋白表达。结果与溃疡性结肠炎组比较,RPA-L组、R-PA-M组、R-PA-H组大鼠结肠组织病理损伤减轻,体质量下降及DAI评分降低(P<0.05);成功分离出大鼠结肠上皮细胞;与NC组比较,Model组IL-1β、TNF-α、IL-6水平、MDA含量、细胞凋亡率、Bax、PARP、cGAS、p-STING、TBK1、IRF3蛋白表达升高,SOD活性降低(P<0.05);与Model组比较,PA-L组、PA-M组、PA-H组IL-1β、TNF-α、IL-6水平、MDA含量、细胞凋亡率、Bax、PARP、cGAS、p-STING、TBK1、IRF3蛋白表达降低,SOD活性升高,RocA组对应指标变化趋势与上述相反(P<0.05);RocA减弱了高剂量PA对溃疡性结肠炎大鼠结肠上皮细胞炎症、氧化应激及细胞凋亡的抑制作用。结论PA可能通过抑制cGAS-STING通路减轻溃疡性结肠炎大鼠结肠上皮细胞损伤。This study was designed to investigate the impact of pachymic acid(PA)on intestinal epithelial cell injury in rats with ulcerative colitis by regulating the cyclic GMP-AMP synthase(cGAS)-stimulator of interferon gene(STING)signal pathway.SD rats were recruited divided into control group,ulcerative colitis group,PA lowdose group(R-PA-L group),PA medium-dose group(R-PA-M group),PA high-dose group(R-PA-H group),RocA(cGAS-STING pathway activator)group(R-RocA group)and R-PA-H+RocA group.Body weight loss and disease activity index(DAI)scores of rats were analyzed;HE staining was applied to detect the pathological changes of colon tissue in rats.The intestinal epithelial cells of rats in the control group and ulcerative colitis group were isolated and identified,then grouped into NC group,Model group,PA low-dose group(PA-L group,2μmol/L),PA medium-dose group(PA-M group,4μmol/L),PA high-dose group(PA-H group,8μmol/L),RocA(cGAS-STING signal pathway activator)group(25 nmol/L)and PA-H+RocA group(8μmol/L+25 nmol/L).The levels of interleukin(IL)-1β,tumor necrosis factor(TNF-α)and IL-6 in cell supernatant were detected by ELISA;the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in rat colon epithelial cells were detected with the kits;the apoptosis of colonic epithelial cells was detected by flow cytometry;Western blot was applied to detect the protein expression of Bcl-2 associated X protein(Bax),poly ADP-ribose polymerase(PARP),cGAS,p-STING,TANK binding kinase 1(TBK1)and interferon regulator factor-3(IRF-3)in rat colon epithelial cells.Compared with ulcerative colitis group,R-PA-L,R-PA-M and R-PA-H groups demonstrated alleviated pathological injury of colonic tissue,diminished body weight loss and lower DAI score(P<0.05).As for the cell experiment,compared with NC group,model group demonstrated higher levels of IL-1β,TNF-α,IL-6,MDA content,apoptosis rate,and higher expression levels of Bax,PARP,cGAS,p-STING,TBK1,IRF-3 proteins,while lower level of SOD activity(P<0.05);compared with model

关 键 词：茯苓酸 环磷酸鸟苷-腺苷酸合成酶-干扰素刺激基因信号通路 溃疡性结肠炎 炎症:氧化应激 凋亡 

分 类 号：R285.5[医药卫生—中药学]