Department of General Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
Department of Chinese Medicine, Dalian Municipal Central Hospital, Dalian, Liaoning 116033, P.R. China.
Mol Med Rep. 2017 Oct;16(4):5121-5128. doi: 10.3892/mmr.2017.7259. Epub 2017 Aug 16.
The present study aimed to evaluate the protective effects of emodin on severe acute pancreatitis (SAP)‑associated acute lung injury (ALI), and investigated the possible mechanism involved. SAP was induced in Sprague‑Dawley rats by retrograde infusion of 5% sodium taurocholate (1 ml/kg), after which, rats were divided into various groups and were administered emodin, FK866 [a competitive inhibitor of pre‑B‑cell colony‑enhancing factor (PBEF)] or dexamethasone (DEX). DEX was used as a positive control. Subsequently, PBEF expression was detected in polymorphonuclear neutrophils (PMNs) isolated from rat peripheral blood by reverse transcription‑quantitative polymerase chain reaction and western blotting. In addition, histological alterations, apoptosis in lung/pancreatic tissues, apoptosis of peripheral blood PMNs and alterations in the expression of apoptosis‑associated proteins were examined by hematoxylin and eosin staining, terminal deoxynucleotidyl‑transferase‑mediated dUTP nick end labeling assay, Annexin V/propidium iodide (PI) assay and western blotting, respectively. Serum amylase activity and wet/dry (W/D) weight ratios were also measured. An in vitro study was also conducted, in which PMNs were obtained from normal Sprague‑Dawley rats and were incubated with emodin, FK866 or DEX in the presence of lipopolysaccharide (LPS). Apoptosis of PMNs and the expression levels of apoptosis‑associated proteins were examined in cultured PMNs in vitro by Annexin V/PI assay and western blotting, respectively. The results demonstrated that emodin, FK866 and DEX significantly downregulated PBEF expression in peripheral blood PMNs. In addition, emodin, FK866 and DEX reduced serum amylase activity, decreased lung and pancreas W/D weight ratios, alleviated lung and pancreatic injuries, and promoted PMN apoptosis by regulating the expression of apoptosis‑associated proteins: Fas, Fas ligand, B‑cell lymphoma (Bcl)‑2‑associated X protein, cleaved caspase‑3 and Bcl‑extra‑large. In addition, the in vitro study demonstrated that emodin, FK866 and DEX significantly reversed the LPS‑induced decrease of apoptosis in PMNs by regulating the expression of apoptosis‑associated proteins. In conclusion, the present study demonstrated that emodin may protect against SAP‑associated ALI by decreasing PBEF expression, and promoting PMN apoptosis via the mitochondrial and death receptor apoptotic pathways.
本研究旨在评估大黄素对重症急性胰腺炎(SAP)相关急性肺损伤(ALI)的保护作用,并探讨其可能的作用机制。通过逆行胰胆管内注入 5%牛磺胆酸钠(1ml/kg)诱导 Sprague-Dawley 大鼠 SAP,之后将大鼠分为不同组,并给予大黄素、FK866(前 B 细胞集落增强因子(PBEF)的竞争性抑制剂)或地塞米松(DEX)治疗。DEX 作为阳性对照。随后,通过逆转录定量聚合酶链反应和蛋白质印迹法检测大鼠外周血中性粒细胞(PMNs)中 PBEF 的表达。此外,通过苏木精和伊红(H&E)染色、末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记(TUNEL)检测、 Annexin V/碘化丙啶(PI)检测和蛋白质印迹法分别检测肺/胰腺组织细胞凋亡、肺和胰腺组织的组织学改变、外周血 PMNs 凋亡以及凋亡相关蛋白的表达变化。还测量了血清淀粉酶活性和湿/干(W/D)重量比。还进行了一项体外研究,其中从正常 Sprague-Dawley 大鼠中获得 PMNs,并在脂多糖(LPS)存在的情况下用大黄素、FK866 或 DEX 孵育 PMNs。通过 Annexin V/PI 检测和蛋白质印迹法分别在体外培养的 PMNs 中检测 PMNs 凋亡和凋亡相关蛋白的表达水平。结果表明,大黄素、FK866 和 DEX 可显著下调外周血 PMNs 中的 PBEF 表达。此外,大黄素、FK866 和 DEX 降低了血清淀粉酶活性,降低了肺和胰腺的 W/D 重量比,减轻了肺和胰腺损伤,并通过调节凋亡相关蛋白:Fas、Fas 配体、B 细胞淋巴瘤(Bcl)-2 相关 X 蛋白、裂解 caspase-3 和 Bcl-extra-large 的表达促进 PMN 凋亡。此外,体外研究表明,大黄素、FK866 和 DEX 通过调节凋亡相关蛋白的表达,显著逆转 LPS 诱导的 PMN 凋亡减少。综上所述,本研究表明,大黄素可能通过降低 PBEF 表达并通过线粒体和死亡受体凋亡途径促进 PMN 凋亡来预防 SAP 相关的 ALI。