基于网络药理学、分子对接及动物实验探讨芍药苷对脓毒症肠损伤的作用及机制  

Study on role and mechanism of paeoniflorin in septic intestinal injury based on network pharmacology,molecular docking and animal experiments

作　　者：雷娇 张铭 龚禹 李若男 谢菁 张彬枫 马玉清[2] LEI Jiao;ZHANG Ming;GONG Yu;LI Ruonan;XIE Jing;ZHANG Binfeng;MA Yuqing(The First Clinical Medical College of Lanzhou University,Lanzhou 730000,China;Department of Anesthesiology,The First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一临床医学院,甘肃兰州730000 [2]兰州大学第一医院麻醉科,甘肃兰州730000

出　　处：《中国病理生理杂志》2025年第3期545-554,共10页Chinese Journal of Pathophysiology

基　　金：甘肃省自然科学基金资助项目(No.24JRRA1075);兰州大学第一医院院内基金(No.ldyyyn2022-5)。

摘　　要：目的:采用网络药理学、分子对接及动物实验结合的方法研究芍药苷(PF)对脂多糖(LPS)诱导的小鼠脓毒症肠损伤的作用及相关机制。方法:网络药理学分析赤芍治疗脓毒症的主要活性成分和作用靶点。分子对接用于探究PF与沉默信息调节因子1(SIRT1)的结合活性。腹腔注射LPS建立小鼠脓毒症肠损伤模型。造模24 h后取材,HE染色观察肠组织病理学变化并进行Chiu's评分;ELISA检测肠组织炎症因子白细胞介素1β(IL-1β)、IL-18、肠通透性指标二胺氧化酶(DAO)、肠型脂肪酸结合蛋白(I-FABP)和血清D-乳酸及有氧糖酵解产物L-乳酸含量;Western blot检测肠组织SIRT1、M2型丙酮酸激酶(PKM2)及NOD样受体蛋白3(NLRP3)蛋白的表达量。结果:网络药理学分析表明赤芍主要活性成分芍药苷可通过作用于SIRT1等靶点治疗脓毒症。分子对接结果显示PF与SIRT1具有较强的结合活性。动物实验结果表明,与对照组比较,LPS组HE染色显示肠组织病理损伤明显,Chiu's评分升高,肠组织IL-1β、IL-18含量升高,DAO、I-FABP含量下降(P<0.05),血清D-乳酸、L-乳酸含量增加(P<0.05),SIRT1表达下调,PKM2、NLRP3表达上调(P<0.05);与LPS组比较,LPS+PF组HE染色和Chiu's评分显示肠组织病理损伤减轻,肠组织IL-1β、IL-18含量下降,DAO、I-FABP含量升高(P<0.05),血清D-乳酸、L-乳酸含量减少(P<0.05),SIRT1表达上调,PKM2、NLRP3表达下调(P<0.05);与LPS+PF组比较,LPS+PF+EX527组HE染色显示肠组织病理损伤加重,Chiu's评分升高,肠组织IL-1β、IL-18含量升高,DAO、I-FABP含量下降(P<0.05),血清D-乳酸、L-乳酸含量增加(P<0.05),SIRT1表达下调,PKM2、NLRP3表达上调(P<0.05)。结论:PF可以缓解小鼠脓毒症肠损伤,其机制可能是通过上调SIRT1表达,抑制PKM2介导的有氧糖酵解进而降低NLRP3活化,减少炎症因子释放,从而减轻肠道炎症反应。AIM:To investigate the effects and underlying mechanisms of paeoniflorin(PF)on lipopolysac-charide(LPS)-induced intestinal injury in septic mice,using a combination of network pharmacology,molecular docking,and animal experiments.METHODS:Network pharmacology was used to identify key active components and therapeutic targets of Red Peony for treating sepsis.Molecular docking was performed to explore the binding affinity be-tween PF and silent information regulator 1(SIRT1).An LPS-induced mouse model of sepsis with intestinal injury was es-tablished.Samples were collected 24 h after modeling,and hematoxylin-eosin(HE)staining was performed to observe pathological changes in intestinal tissues.Chiu's scoring system was utilized to evaluate the extent of intestinal injury.Enzyme-linked immunosorbent assay(ELISA)was employed to measure levels of inflammatory factors in intestinal tissues,including interleukin-1β(IL-1β)and IL-18,as well as indicators of intestinal permeability such as diamine oxidase(DAO)and intestinal-type fatty acid-binding protein(I-FABP),alongside serum levels of D-lactate and the aerobic gly-colysis product L-lactate.Western blot analysis was performed to assess changes in protein levels of SIRT1,M2-type pyru-vate kinase(PKM2),and NOD-like receptor protein 3(NLRP3)in intestinal tissues.RESULTS:Network pharmacolo-gy suggested that paeoniflorin,an active component of Red Peony,treats sepsis by targeting SIRT1 among other proteins.Molecular docking revealed a strong binding affinity of PF with SIRT1.In vivo experimentation revealed significant patho-logical damage in intestinal tissues in the LPS group compared to the control group as evidenced by HE staining.Chiu's score,along with levels of IL-1β,IL-18,D-lactate,and L-lactate were significantly elevated,while DAO and I-FABP levels were reduced(P<0.05).SIRT1 expression decreased,while PKM2 and NLRP3 levels increased(P<0.05).In contrast,the LPS+PF group displayed reduced intestinal histopathological injury,lower Chiu's scores,and decreased levels o

关 键 词：脓毒症 肠损伤 芍药苷 沉默信息调节因子1 M2型丙酮酸激酶 

分 类 号：R631.3[医药卫生—外科学] R282.71[医药卫生—临床医学] R574