ROS-NLRP3信号通路在急性胰腺炎大鼠中的作用机制研究  

Mechanism of ROS-NLRP3 signaling pathway in rats with acute pancreatitis

作　　者：陈金凤 蒙诺 雷宇[1] 冯勇[1] 李旺健 唐曦平 CHEN Jin‑feng;MENG Nuo;LEI Yu;FENG Yong;LI Wang‑jian;TANG Xi‑ping(Guangxi Medical University,Nanning 530200,China;Guangxi Medical University Cancer Hospital,Nanning 530021,China)

机构地区：[1]广西医科大学,广西南宁530200 [2]广西医科大学附属肿瘤医院,广西南宁530021

出　　处：《海南医学院学报》2023年第2期81-86,95,共7页Journal of Hainan Medical University

基　　金：广西自然科学基金面上项目(2020GXNSFAA297062);广西卫生适宜推广项目(S2020103);广西中医药管理局立项项目(GZZC2020235)。

摘　　要：目的:探讨抑制ROS-NLRP3信号通路对急性胰腺炎大鼠的保护作用以及相关分子机制。方法:将36只雄性SD大鼠随机分为对照组(NC组)、轻症急性胰腺炎模型组(AP组)、轻症急性胰腺炎+N-乙酰半胱氨酸干预组(AP+NAC组)、假手术组(SO组)、重症急性胰腺炎模型组(SAP组),重症急性胰腺炎+N-乙酰半胱氨酸干预组(SAP+NAC组),每组6只。模型构造24 h后处死大鼠。苏木精-伊红(HE)染色观察大鼠胰腺组织病理改变并进行病理评分;二氢乙锭(DHE)荧光探针检测大鼠胰腺腺泡细胞ROS水平;WST-1法检测大鼠血清超氧化物歧化酶(SOD)水平;免疫组化法(IHC)检测大鼠胰腺组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)蛋白表达;采用qPCR检测SO组、SAP组、SAP+NAC组肾脏组织中NLRP3 mRNA水平;采用ELISA检测血清中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的水平。结果:与对照组比较,模型组胰腺组织病理学评分、ROS水平、NLRP3蛋白水平、肾脏组织NLRP3 mRNA水平、血清IL-1β和TNF-α含量明显升高(P<0.05),模型组SOD活性降低(P<0.05);与模型组相比,NAC干预组胰腺组织病理学评分、ROS水平、NLRP3蛋白水平、肾脏组织NLRP3的mRNA水平、血清IL-1β和TNF-α含量降低(P<0.05),SOD活性升高(P<0.05)。结论:抑制ROS-NLRP3信号通路可以减轻大鼠AP模型及SAP模型的胰腺损伤,降低SAP模型大鼠肾脏组织炎性水平。Objective:To investigate the protective effect of inhibiting ROS-NLRP3 signaling pathway on acute pancreatitis in rats and the related molecular mechanism.Methods:Thirty-six male SD rats were randomly divided into control group(NC group),mild acute pancreatitis model group(AP group),and mild acute pancreatitis+N-acetylcysteine intervention group(AP+NAC group),sham operation group(SO group),severe acute pancreatitis model group(SAP group)and severe acute pancreatitis+N-acetylcysteine intervention group(SAP+NAC group)with six rats in each group.The rats were sacrificed 24 h after establishing the models.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of rat pancreas and make pathological scores.The level of ROS in rat pancreatic acinar cells was detected by dihydroethidium(DHE)fluorescent probe.The level of superoxide dismutase(SOD)in serum was detected by WST-1 method;the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)protein in rat pancreas was detected by immunohistochemistry(IHC).The levels of NLRP3 mRNA in kidney tissue of SO group,SAP group and SAP+NAC group were detected by qPCR;the levels of interleukin-1β(IL-1β)and tumor necrosis factorα(TNF-α)in serum were detected by ELISA.Results:Compared with the control group,the pancreas histopathological score,ROS level,NLRP3 protein level,kidney tissue NLRP3 mRNA level,and serum IL-1βand TNF-αlevels in the model group were significantly increased(P<0.05),and the SOD activity in the model group was decreased(P<0.05).And compared with the model group,the pancreatic histopathological score,ROS level,NLRP3 protein level,NLRP3 mRNA level in kidney tissue,and serum IL-1βand TNF-αlevels in the NAC intervention group decreased(P<0.05),and the SOD activity increased(P<0.05).Conclusions:Inhibition of ROS-NLRP3 signaling pathway can alleviate pancreatic injury in rat AP model and SAP model,and reduce the level of renal tissue inflammation in SAP model rats.

关 键 词：活性氧 NOD样受体蛋白3 急性胰腺炎 

分 类 号：R576[医药卫生—消化系统]