Aqueous extraction from dachengqi formula granules reduces the severity of mouse acute pancreatitis via inhibition of pancreatic pro-inflammatory signalling pathways
Aqueous extraction from dachengqi formula granules reduces the severity of mouse acute pancreatitis via inhibition of pancreatic pro-inflammatory signalling pathways
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大承气配方颗粒的水提取物通过抑制胰腺促炎信号通路降低小鼠急性胰腺炎的严重程度
DOI：
10.1016/j.jep.2020.112861
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发表时间：
2020
影响因子：
5.4
通讯作者：
Xia Qing
中科院分区：
文献类型：
--
作者：
Ma Xiaohua;Jin Tao;Han Chenxia;Shi Na;Liang Ge;Wen Yongjian;Yang Jingyu;Fu Xianghui;Lan Tian;Jiang Kun;Nunes Quentin M.;Chvanov Michael;Criddle David N.;Philips Anthony R.;Deng Lihui;Liu Tingting;Windsor John A.;Sutton Robert;Du Dan;Huang Wei;Xia Qing
丹承气汤（DCQD）是中国广泛用于治疗急性胰腺炎（AP）的泻性中药。AP是一种常见的消化系统疾病，目前没有有效的药物干预。配方颗粒由于其效力保留，剂量精确和易于使用的优点，已成为中国草药配方递送的首选方法。DCQD配方颗粒（DFGs）对实验性AP模型的疗效尚未研究。目的分析和比较DFGs及其水萃取衍生物（AE）和氯仿萃取衍生物（CE）的化学成分差异。为了评估其对新分离的腺泡细胞和两种实验性ap模型的严重程度和靶向胰腺促炎信号通路的影响。材料和方法采用suplc - q - tof - ms分析了DFGs及其提取物的化学成分。新分离的小鼠胰腺腺泡细胞用牛磺酸胆酸3-硫酸二钠盐（TLCS, 500 μM）处理，有或没有DFGs、AE和CE。凋亡和坏死细胞死亡途径的激活分别用caspase 3/7 （10 μl/mL）和碘化丙啶（PI, 1 μM），荧光板阅读器检测。荧光显微镜下观察坏死腺泡细胞计数。小鼠每小时腹腔注射7次紫蛋白（50 μg/kg）或逆行注射TLCS （3 mM, 50 μl）诱导AP （CER-AP和TLCS-AP）。在er - ap中，小鼠口服DFGs（2.1、4.2和5.2 g/kg）、AE（0.6、1.2和2.4 g/kg）和CE(4,9和17 mg/kg)，或匹配DFGs （1.8 g/kg）和AE (1 g/kg)，每2小时间隔3次，或在第3次注射小檗碱时开始单次腹腔注射dcqd相关单体大黄碱（20 mg/kg）、narigeinine （25 mg/kg）和宏木酚（5 mg/kg）。TLCS-AP组分别于术后1、3、5 h口服DFGs （4.2 g/kg）。测定疾病严重程度和胰腺促炎标志物。结果发现了主要有效的蒽醌类及其苷类、黄酮类及其苷类、多酚类和木脂素类化合物。蒽醌类苷类、酚类、黄酮类等极性成分含量较高。相反，含有甲氧基取代黄酮类化合物和蒽醌类化合物的低极性组分在CE中更丰富。以4.2 g/kg剂量给予DFGs，在CER-AP和TLCS-AP中观察到胰腺组织病理学评分和严重程度指数的一致降低。在体外，AE显著降低了凋亡和坏死细胞死亡通路的激活，而CE则增加了tlcs诱导的腺泡细胞坏死。在体内，1.2 g/kg剂量的AE持续降低胰腺组织病理学评分和CER-AP中的髓过氧化物酶，这与抑制促炎介质mrna和蛋白质的表达有关。CE增加了肺髓过氧化物酶，在所有剂量下都不能预防CER-AP。AE在降低胰腺组织病理学评分和髓过氧化物酶方面比DFGs更有效。结论DFGs中的sae通过抑制胰腺促炎信号通路减轻小鼠AP模型的严重程度。AE对实验AP的作用强于其原始的DFGs和DCQD单体。
Ethnopharmacological relevanceDachengqi decoction (DCQD) belongs to a family of purgative herbal formulas widely used in China for the treatment of acute pancreatitis (AP). AP is a prevalent digestive disease currently without an effective pharmacological intervention. Formula granules have become the preferred method for delivery of herbal formulation in China given its benefit of potency retention, dosing precision and ease of use. The efficacy of DCQD formula granules (DFGs) in experimental AP models has not been investigated.Aim of the studyTo analyse and compare the differences in chemical composition of DFGs, with their aqueous extraction (AE) and chloroform extraction (CE) derivatives. To assess their efficacy on severity and targeted pancreatic pro-inflammatory signalling pathways in freshly isolated acinar cells and two models of experimental AP.Material and methodsUPLC-Q-TOF-MS was used to analyse chemical components of DFGs and their extractions. Freshly isolated mouse pancreatic acinar cells were treated with taurolithocholic acid 3-sulphate disodium salt (TLCS, 500 μM) with or without DFGs, AE and CE. Apoptotic and necrotic cell death pathway activation was measured by caspase 3/7 (10 μl/mL) and propidium iodide (PI, 1 μM), respectively, using a fluorescent plate reader. Necrotic acinar cells were also counted by epifluorescence microscopy. Mice received either 7 intraperitoneal injections of caerulein (50 μg/kg) at hourly intervals or retrograde infusion of TLCS (3 mM, 50 μl) to induce AP (CER-AP and TLCS-AP, respectively). In CER-AP, mice received oral gavage of DFGs (2.1, 4.2 and 5.2 g/kg), AE (0.6, 1.2, and 2.4 g/kg) and CE (4, 9 and 17 mg/kg), or matched DFGs (1.8 g/kg) and AE (1 g/kg) for 3 times at 2-hourly intervals, or a single intraperitoneal injection of DCQD-related monomers rhein (20 mg/kg), narigeinine (25 mg/kg), and honokiol (5 mg/kg) begun at the 3rd injection of caerulein. In TLCS-AP, DFGs (4.2 g/kg) were given orally at 1, 3 and 5 h post-surgery. Disease severity and pancreatic pro-inflammatory markers were determined.ResultsThe main effective anthraquinones and their glycosides, flavonoids and their glycosides, polyphenols and lignans were found in the DFGs. A higher proportion of polar components including glycosides attached to anthraquinones, phenols and flavonoids was found in AE. Conversely, lower polar components containing methoxy substituted flavonoids and anthraquinones were more abundant in CE. DFGs were given at 4.2 g/kg, a consistent reduction in the pancreatic histopathology score and severity indices was observed in both CER-AP and TLCS-AP.In vitro, AE significantly reduced both apoptotic and necrotic cell death pathway activation, while CE increased TLCS-induced acinar cell necrosis.In vivo, AE at dose of 1.2 g/kg consistently reduced pancreatic histopathological scores and myeloperoxidase in the CER-AP that were associated with suppressed expression of pro-inflammatory meditator mRNAs and proteins. CE increased lung myeloperoxidase and failed to protect against CER-AP in all dosages. AE was demonstrated to be more effective than DFGs in reducing pancreatic histopathological scores and myeloperoxidase.ConclusionsAE from DFGs alleviated the severity of mouse AP models via an inhibition of pancreatic pro-inflammatory signalling pathways. Efficacy of AE on experimental AP was more potent than its original DFGs and DCQD monomers.