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

龙润; 许亚培; 杨铸锋; 杨静; 李艳红

doi:10.16098/j.issn.0529-1356.2021.04.019

PDF(4532 KB)

解剖学报 ›› 2021, Vol. 52 ›› Issue (4) : 628-634. DOI: 10.16098/j.issn.0529-1356.2021.04.019

组织学胚胎学发育生物学

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

龙润¹ 许亚培¹ 杨铸锋¹ 杨静¹ 李艳红^2*

作者信息 +

Effects of ginsenoside Re on intestinal mucosa of mice with severe acute pancreatitis and its regulation mechanism

LONG Run¹ XU Ya-pei¹ YANG Zhu-feng¹ YANG Jing¹ LI Yan-hong^2*

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2021.04.019

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 https://doi.org/10.16098/j.issn.0529-1356.2021.04.019

中图分类号： R576

参考文献

［1］ Portelli M, Jones CD. Severe acute pancreatitis: pathogenesis, diagnosis and surgical management［J］. Hepatobiliary Pancreat Dis Int, 2017, 16(2):155-159.

［2］ Su JL, Huang Zh, Sun HY, et al. Impact of timing of abdominal paracentesis drainage on treatment outcomes in patients with severe acute pancreatitis［J］. Chinese Journal of Hepatobiliary Surgery, 2018, 24(10): 692-697. (in Chinese)

苏江林, 黄竹, 孙红玉, 等. 腹腔引流穿刺时机对重症急性胰腺炎患者预后的影响［J］. 中华肝胆外科杂志, 2018, 24(10): 692-697

［3］ Wang MJ, Fang XP, Li XY, et al. Study on the relationship between early gastric and intestinal function and disease progression in patients with acute pancreatitis［J］. Chinese General Practice, 2019, 22(1): 59-62. (in Chinese)

王旻静, 方小萍, 李雪阳, 等. 急性胰腺炎患者住院早期胃小肠功能与病情进展的相关性研究［J］. 中国全科医学, 2019, 22(1): 59-62.

［4］ Pan X, Fang X, Wang F, et al. Butyrate ameliorates caerulein-induced acute pancreatitis and associated intestinal injury by tissue-specific mechanisms［J］. Br J Pharmacol, 2019, 176(23): 4446-4461.

［5］ Dong J, Chen X, Song Y, et al. Chaiqin Chengqi decoction inhibits inflammatory mediators and attenuates acute pancreatitis through deactivation of Janus kinase/signal transducer and activator of transcription signaling pathway［J］. J Tradit Chin Med, 2019, 39(2): 166-173.

［6］ Richmond CA, Rickner H, Shah MS, et al. JAK/STAT-1 signaling is required for reserve intestinal stem cell activation during intestinal regeneration following acute inflammation［J］. Stem Cell Reports, 2018, 10(1): 17-26.

［7］ Wang QH, Shang QQ, Zhang JM, et al. Study of ginsenoside compound K on the effect of immunity and liver function in rats with severe acute pancreatitis［J］. Journal of Binzhou Medical University, 2018, 41(4): 241-243, 258. (in Chinese)

王庆华, 商琼琼, 张建民, 等. 人参皂苷CK对重症急性胰腺炎大鼠肝功能和免疫作用的影响［J］. 滨州医学院学报, 2018, 41(4): 241-243, 258.

［8］ Leng X, Song N, Zang AY, et al. Role of JAK2/STAT3 signaling pathway in ginsenoside Re pretreatment in prevention of isoproterenol-induced acute myocardial ischemia［J］. Chinese Pharmacological Bulletin, 2018, 34(1): 103-107. (in Chinese)

冷雪, 宋囡, 臧安缘, 等. JAK2/STAT3信号通路在人参皂苷Re预处理预防异丙肾上腺素致急性心肌缺血损伤中的作用［J］. 中国药理学通报, 2018, 34(1): 103-107.

［9］ He DM, Yang TZh, Liu Y, et al. The role of P-selectin glycoprotein ligand 1 in the pathogenesis of acute severe pancreatitis［J］. Chinese Journal of Experimental Surgery, 2020, 37(3): 477-479. (in Chinese)

何冬梅, 杨铁柱, 刘燕, 等. P选择素糖蛋白配-1在急性重症胰腺炎发生中的作用［J］. 中华实验外科杂志, 2020, 37(3): 477-479.

［10］ Li JY, Wang Zh, Liu Y, et al. Protective effect of ginsenoside Re on biomarkers in mice with Alzheimer’s disease［J］. Practical Geriatrics, 2017, 31(10): 921-926. (in Chinese)

李菁媛, 王喆, 刘颖, 等. 人参皂苷Re对阿尔茨海默病模型小鼠脑组织生物标记物调控作用的研究［J］. 实用老年医学, 2017, 31(10): 921-926.

［11］ Ma L, Clayton JR, Walgren RA, et al. Discovery and characterization of LY2784544, a small-molecule tyrosine kinase inhibitor of JAK2V617F［J］. Blood Cancer J, 2013, 3(4): 109-113.

［12］ Ren YF, Wang MZ, Bi JB, et al. Irisin attenuates intestinal injury, oxidative and endoplasmic reticulum stress in mice with L-arginine-induced acute pancreatitis［J］. World J Gastroenterol, 2019, 25(45): 6653-6667.

［13］ Yang GR, Yang ChCh. Application progress of Shenfu Injection in critically ill patients［J］. Pharmaceutical Journal of Chinese People’s Liberation Army, 2017, 33(3): 259-261. (in Chinese)

杨贵荣, 杨长春. 参附注射液在危重病中的应用进展［J］. 解放军药学学报, 2017, 33(3): 259-261.

［14］ Li M, Zhang X, Wang B, et al. Effect of JAK2/STAT3 signaling pathway on liver injury associated with severe acute pancreatitis in rats［J］. Exp Ther Med, 2018, 16(3): 2013-2021

［15］ Wu AX. Effect of astragaloside Ⅳ on pancreas of mice with severe acute pancreatitis［J］. Chinese Journal of Clinical Pharmacology, 2019, 35(18): 2065-2067. (in Chinese)

吴爱祥. 黄芪甲苷对重症急性胰腺炎小鼠胰腺的影响［J］. 中国临床药理学杂志, 2019, 35(18): 2065-2067.

［16］ Lou YJ, Zhang QCh. The progress of molecular biology’s mechanism about inhibiting effect of ginsenoside to gastration-resistant prostat cancer［J］.Journal of Modern Oncology, 2018, 245(11): 156-160. (in Chinese)

楼雨蛟, 张青川. 人参皂苷抑制去势抵抗性前列腺癌的分子生物学机制［J］. 现代肿瘤医学, 2018, 245(11): 156-160.

［17］ Tang Q, Wang H, Wang X, et al. Effect of PI3K/PKB signal pathway inhibitor wortmannin pretreatment on intestinal barrier function in severe acute pancreatitic rats［J］. Adv Clin Exp Med, 2019, 28(8): 1059-1066.

［18］ Xiao Y, Jin D. Research progress on mechanism of ginsenoside improving inflammatory bowel disease［J］. Chinese Journal of Immunology, 2020, 3(3): 112-115. (in Chinese)

肖瑶, 金丹. 人参及人参皂苷改善炎症性肠病免疫机制相关的研究进展［J］. 中国免疫学杂志, 2020, 3(3): 112-115.

［19］ Li MM, Cui HL. Effects of ginsenoside on atrophy of intestinal mucosa and expressions of CRP, TNF-α and IL-6 in rats with severe abdominal infection［J］. Jiangsu Medical Journal, 2019, 45(2): 2, 11-13. (in Chinese)

李苗苗, 崔华雷. 人参皂苷对重症腹腔感染大鼠肠黏膜萎缩及CRP、TNF-α和IL-6表达的影响［J］. 江苏医药, 2019, 45(2): 2, 11-13.

［20］ Miao ZhW, Yan J, Gu MJ, et al. The effects of ginsenoside Rg3 on the Th1/Th2 imbalance in DSS induced colitis mice［J］. Pharmacology and Clinics of Chinese Materia Medica, 2019, 35(1): 47-51. (in Chinese)

缪志伟, 严晶, 顾鸣佳, 等. 人参皂苷Rg3对DSS诱导的溃疡性结肠炎小鼠Th1/Th2失衡的影响［J］. 中药药理与临床, 2019, 35(1): 47-51.