人参皂苷Re对重症急性胰腺炎小鼠小肠黏膜的影响及调控机制

doi:10.16098/j.issn.0529-1356.2021.04.019

摘要/Abstract

摘要：

目的探讨人参皂苷Re通过调控 Janus激酶2/信号转导子与转录激活子3（JAK2/STAT3）通路对重症急性胰腺炎（SAP）小鼠小肠黏膜的影响及作用机制。方法 48只小鼠分为对照组、SAP组、SAP+人参皂苷Re组和SAP+人参皂苷Re+LY2784544组（n=12）。通过腹腔注射雨蛙肽溶液（禁食12 h后，100 μg/kg × 6次，注射间隔60 min）的方法建立SAP模型。腹腔注射人参皂苷Re（4 mg/kg，每日1次，连续7 d）。通过腹腔注射 12.7 mg/kg LY2784544来抑制JAK2/STAT3通路。检测各组胰腺指数、血清淀粉酶和炎性因子水平；检测D-乳酸和FITC-D水平分析小肠黏膜屏障功能；HE染色观察胰腺组织和小肠黏膜组织损伤情况；Western blotting检测小肠黏膜组织中凋亡蛋白Bax和Bcl-2的水平；Real-time PCR和Western blotting检测胰腺组织和小肠黏膜组织JAK2/STAT3通路水平。结果 SAP组小鼠胰腺指数、血清淀粉酶、白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）、FITC-D、D-乳酸、Bax蛋白水平、胰腺组织和小肠组织中JAK2和STAT3 mRNA和蛋白水平均显著高于对照组，而Bcl-2蛋白显著低于对照组（P<0.05）。SAP+人参皂苷Re组小鼠Bcl-2蛋白显著高于SAP组，其余指标显著低于SAP组（P<0.05）。SAP+人参皂苷Re+LY2784544组的Bcl-2蛋白显著高于SAP+人参皂苷Re组，其余指标显著低于SAP+人参皂苷Re组（P<0.05）。结论人参皂苷Re可能通过抑制JAK/STAT通路缓解炎性反应减轻SAP模型小鼠的胰腺损伤, 并通过抑制细胞的凋亡保护小肠黏膜屏障。

关键词: 重症急性胰腺炎, 人参, 人参皂苷Re, JAK2/STAT3通路, 实时定量聚合酶链反应, 免疫印迹法, 小鼠

Abstract:

Objective To explore the effect and mechanism of ginsenoside Re on the intestinal mucosa of severe acute pancreatitis (SAP) mice by regulating the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) pathway. Methods Totally 48 mice were divided into: control group, SAP group, SAP + ginsenoside Re group and SAP+ginsenoside Re+LY2784544 group (n=12). The mice were intraperitoneally injected with caerulein solution (after fasting for 12 hours, 100 μg/kg, 6 times, injection interval 60 minutes) to establish SAP models. Mice in the SAP+ginsenoside Re group were intraperitoneally injected with ginsenoside Re (4 mg/kg, 1 time a day for 7 consecutive days). Intraperitoneal injection of 12.7 mg/kg LY2784544 was used to inhibit the JAK2/STAT3 pathway. Pancreatic and intestinal mucosal injury were detected in each group. The wet to dry weight ratio of pancreas, serum amylase and inflammatory factor levels were detected in each group. The intestinal mucosal barrier function was analyzed by detecting the levels of D-lactic acid and fluorescein isothiocyanate dextran (FITC-D). The damage of pancreatic tissue and intestinal mucosa tissue was observed by HE staining. Western blotting was used to detect the levels of apoptotic proteins Bax and Bcl-2 in the intestinal mucosa. The JAK2/STAT3 pathway expression levels in pancreatic tissue and intestinal mucosa tissue were detected by Real-time PCR and Western blotting. Results The pancreatic index, serum amylase, interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α), FITC-D, D-lactic acid, Bax protein levels, JAK2 and STAT3 mRNA and protein levels in the SAP group were significantly higher than those in the control group, while Bcl-2 protein was significantly lower than that in the control group (P<0.05). The Bcl-2 protein of SAP+ginsenoside Re group was significantly higher than that of SAP group, and other indexes were significantly lower than those in SAP group (P<0.05). The Bcl-2 protein of SAP+ginsenoside Re+LY2784544 group was significantly higher than that of SAP+ginsenoside Re group, and other indexes were significantly lower than those in the SAP+ginsenoside Re group (P<0.05). Conclusion Ginsenoside Re may reduce the pancreatic injury in SAP model mice by inhibiting the JAK/STAT pathway to alleviate the inflammatory response, and may protect the small intestinal mucosal barrier by alleviating pancreatitis and inhibiting the intestinal mucosal JAK/STAT pathway to inhibit cell apoptosis.

Key words: Severe acute pancreatitis, Ginseng, Ginsenoside Re, JAK2/STAT3 pathway, Real-time PCR, Western blotting, Mouse

中图分类号:

R576

龙润许亚培杨铸锋杨静李艳红. 人参皂苷Re对重症急性胰腺炎小鼠小肠黏膜的影响及调控机制[J]. 解剖学报, 2021, 52(4): 628-634.

LONG Run XU Ya-pei YANG Zhu-feng YANG Jing LI Yan-hong. Effects of ginsenoside Re on intestinal mucosa of mice with severe acute pancreatitis and its regulation mechanism[J]. Acta Anatomica Sinica, 2021, 52(4): 628-634.

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