大黄素干预在大鼠重症急性胰腺炎肠神经损伤中的作用及相关机制  

Effect and mechanism of emodin intervention on intestinal nerve injury in rats with severe acute pancreatitis

作　　者：赵楠 代佳玲 丁勇 许宝珠 杨丽[1] 陈娟 梁国刚[2] Zhao Nan;Dai Jialing;Ding Yong;Xu Baozhu;Yang Li;Chen Juan;Liang Guogang(State Key Discipline Laboratory of Clinical Integrated Traditional Chinese and Western Medicine,the First Affiliated Hospital of Dalian Medical University,Dalian 116000,China;Department of General SurgeryⅢ,the First Affiliated Hospital of Dalian Medical University,Dalian 116000,China)

机构地区：[1]大连医科大学附属第一医院中西医结合临床国家重点学科实验室,大连116000 [2]大连医科大学附属第一医院普外三科,大连116000

出　　处：《中华肝胆外科杂志》2024年第9期684-690,共7页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金(81873164)。

摘　　要：目的探讨大黄素通过核苷酸结合寡聚化结构域样受体蛋白3/含半胱氨酸的天冬氨酸蛋白水解酶-1(NLRP3/Caspase-1)通路在大鼠重症急性胰腺炎(SAP)肠神经炎中的作用及大黄素的干预效果。方法选取6~8周龄体重200 g左右的健康雄性SD大鼠40只,随机分为对照组、SAP模型组、大黄素干预(EMO)组、NLRP3敲低组。利用胰胆管逆行注射去氧胆酸钠诱导大鼠SAP模型并检测血清淀粉酶;筛选有效NLRP3敲低序列用于NLRP3敲低组动物实验;荧光定量聚合酶链反应检测各组大鼠小肠NLRP3、Caspase-1、白细胞介素(IL)-1β、IL-18、肿瘤坏死因子-α(TNF-α)、加斯特明-D(GSDMD)的基因表达变化。免疫荧光检测各组小肠胶质纤维酸性蛋白(GFAP)的表达。结果对照组、SAP组、EMO组、NLRP3敲低组大鼠的淀粉酶水平分别为(277.73±24.92)U/L、(1018.57±282.89)U/L、(625.43±134.40)U/L、(391.01±27.63)U/L,其中SAP组、EMO组高于对照组,EMO组、NLRP3敲低组低于SAP组,差异均具有统计学意义(均P<0.001)。与对照组相比,SAP组的NLRP3、Caspase-1、IL-1β、IL-18、TNF-α、GSDMD的基因表达水平均增加,差异均有统计学意义(均P<0.001);与SAP组相比,NLRP3敲低组的上述6个指标基因表达水平皆减少,EMO组的NLRP3、IL-1β、IL-18、TNF-α基因表达亦减少,差异均具有统计学意义(均P<0.05)。对照组、SAP组、EMO组、NLRP3敲低组大鼠小肠的GFAP相对表达分别为(1.00±0)、(1.66±0.11)、(1.13±0.02)、(1.13±0.02),其中与对照组相比SAP组的GFAP表达增加,而EMO组和NLRP3敲低组的GFAP表达低于模型组,差异均具有统计学意义(均P<0.05)。结论大黄素及敲低NLRP3均可通过NLRP3/Caspase-1信号通路促进SAP小肠损伤修复,进而起到肠保护作用。ObjectiveTo explore the role and the intervention effect of emodin in intestinal neuropathy in rats with severe acute pancreatitis(SAP)through the nucleotide binding oligomerization domain like receptor protein 3/cysteine containing aspartic acid protease-1(NLRP3/Caspase-1)pathway.MethodsForty male healthy SD rats aged 6-8 weeks with a weight of approximately 200g were randomly divided into control group,SAP model group,emodin treatment(EMO)group,and NLRP3 knockdown group.SAP were induced by retrograde injection of sodium deoxycholate into the pancreatic duct of rats and serum amylase of which were detected.The effective NLRP3 knockdown sequence was screened for NLRP3 knockdown animal experiments.Fluorescence quantitative polymerase chain reaction was used to detect the expression of NLRP3,Caspase-1,gasdermin-D(GSDMD),interleukin(IL)-1β,IL-18 and tumor necrosis factor-α(TNF-α)in the small intestine of each group.Immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP)in the small intestine of each group.ResultsThe amylase levels of the control group,SAP group,EMO group,and NLRP3 knockdown group were(277.73±24.92)U/L,(1018.57±282.89)U/L,(625.43±134.40)U/L,and(391.01±27.63)U/L,respectively.The SAP and EMO groups were significantly higher than the control group(P<0.001),while the EMO and NLRP3 knockdown groups were significantly lower than the SAP group(all P<0.001).Compared with control group,the expression levels of NLRP3,Caspase-1,IL-1β,IL-18,TNF-αand GSDMD in SAP group were increased,with statistical significance(all P<0.001).Compared with SAP group,the NLRP3 knockdown group showed the expressionlevels of the above 6 genes were all decreased,and EMO group showed decreased gene expressing levels of NLRP3,IL-1β,IL-18 and TNF-α,with statistical significance(all P<0.05).The relative expression of GFAP in small intestine of control group,SAP group,EMO group and NLRP3 knockdown group were(1.00±0),(1.66±0.11),(1.13±0.02)and(1.13±0.02),respectively.Among them,

关 键 词：胰腺炎 重症急性 大黄素 肠神经胶质细胞 肠神经炎 慢病毒 

分 类 号：R576[医药卫生—消化系统]