|
1
|
Jiang H, Xie W, Chen Q, Li Y, Yu Z, Liu N. Construction and validation of a rat model of acute necrotizing pancreatitis-associated intestinal injury. Am J Physiol Gastrointest Liver Physiol 2024; 327:G80-G92. [PMID: 38742280 PMCID: PMC11376975 DOI: 10.1152/ajpgi.00262.2023] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/07/2023] [Revised: 04/22/2024] [Accepted: 05/11/2024] [Indexed: 05/16/2024]
Abstract
Acute pancreatitis (AP) is an acute inflammatory reaction of the pancreatic tissue, which involves auto-digestion, edema, hemorrhage, and necrosis. AP can be categorized into mild, moderately severe, and severe AP, with severe pancreatitis also referred to as acute necrotizing pancreatitis (ANP). ANP is characterized by the accumulation of necrotic material in the peritoneal cavity. This can result in intestinal injury. However, the mechanism of ANP-associated intestinal injury remains unclear. We established an ANP-associated intestinal injury rat model (ANP-IR model) by injecting pancreatitis-associated ascites fluid (PAAF) and necrotic pancreatic tissue at various proportions into the triangular area formed by the left renal artery and ureter. The feasibility of the ANP-IR model was verified by comparing the similar changes in indicators of intestinal inflammation and barrier function between the two rat models. In addition, we detected changes in apoptosis levels and YAP protein expression in the ileal tissues of rats in each group and validated them in vitro in rat epithelial crypt cells (IEC-6) to further explore the potential injury mechanisms of ANP-associated intestinal injury. We also collected clinical data from patients with ANP to validate the effects of PAAF and pancreatic necrosis on intestinal injury. Our findings offer a theoretical basis for restricting the buildup of peritoneal necrosis in individuals with ANP, thus promoting the restoration of intestinal function and enhancing treatment efficacy. The use of the ANP-IR model in further studies can help us better understand the mechanism and treatment of ANP-associated intestinal injury.NEW & NOTEWORTHY We constructed a rat model of acute necrotizing pancreatitis-associated intestinal injury and verified its feasibility. In addition, we identified the mechanism by which necrotic pancreatic tissue and pancreatitis-associated ascites fluid (PAAF) cause intestinal injury through the HIPPO signaling pathway.
Collapse
Affiliation(s)
- Haojie Jiang
- Wenzhou Medical University, Wenzhou, People's Republic of China
- Department of Gastrointestinal Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People's Republic of China
- Zhejiang Key Laboratory of Intelligent Cancer Biomarker Discovery and Translation, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, People's Republic of China
| | - Weidong Xie
- Wenzhou Medical University, Wenzhou, People's Republic of China
- Department of Gastrointestinal Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People's Republic of China
| | - Qinbo Chen
- Department of Gastrointestinal Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People's Republic of China
- Zhejiang Key Laboratory of Intelligent Cancer Biomarker Discovery and Translation, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, People's Republic of China
| | - Yiling Li
- Wenzhou Medical University, Wenzhou, People's Republic of China
- Department of Gastrointestinal Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People's Republic of China
| | - Zhen Yu
- Wenzhou Medical University, Wenzhou, People's Republic of China
- Department of Gastrointestinal Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People's Republic of China
- Shanghai Tenth People's Hospital, Shanghai, People's Republic of China
| | - Naxin Liu
- Wenzhou Medical University, Wenzhou, People's Republic of China
- Department of Gastrointestinal Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People's Republic of China
| |
Collapse
|
|
2
|
Yuan X, Luo C, Wu J, Li W, Guo X, Li S, Wang B, Sun H, Tang L. Abdominal paracentesis drainage attenuates intestinal mucosal barrier damage through macrophage polarization in severe acute pancreatitis. Exp Biol Med (Maywood) 2021; 246:2029-2038. [PMID: 34053233 PMCID: PMC8474980 DOI: 10.1177/15353702211015144] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/13/2020] [Accepted: 04/12/2021] [Indexed: 11/15/2022] Open
Abstract
Abdominal paracentesis drainage (APD), as an effective treatment of severe acute pancreatitis (SAP) in clinical settings, can ameliorate intestinal barrier damage and the overall severity of SAP. However, the mechanism underlying therapeutic effects of APD on damaged intestinal mucosal barrier during SAP is still unclear. Here, SAP was induced by injecting 5% Na-taurocholate retrograde into the biliopancreatic duct of rats to confirm the benefits of APD on enteral injury of SAP and further explore the possible mechanism. Abdominal catheter was placed after SAP was induced in APD group. As control group, the sham group received no operation except abdominal opening and closure. By comparing changes among control group, sham group, and APD group, APD treatment obviously lowered the intestinal damage and reduced the permeation of intestinal mucosal barrier, which was evidenced by intestinal H&E staining, enteral expression of tight junction proteins, intestinal apoptosis measurement and detection of serum diamine oxidase, intestinal fatty acid binding protein and D-lactic acid. Furthermore, we found that APD polarized intestinal macrophages toward M2 phenotype by the determination of immunofluorescence and western blotting, and this accounts for the benefits of APD for intestinal injury in SAP. Importantly, the protective effect against intestinal injury by APD treatment was mediated through the inhibited ASK1/JNK pathway. In summary, APD improved the intestinal mucosal barrier damage in rats with SAP through an increasing portion of M2 phenotype macrophages in intestine via inhibiting ASK1/JNK pathway.
Collapse
Affiliation(s)
- Xiaohui Yuan
- College of Medicine, Southwest Jiaotong University, Chengdu
610031, China
- Department of General Surgery & Pancreatic Injury and Repair
Key Laboratory of Sichuan Province, The General Hospital of Western Theater
Command, Chengdu 610083, China
| | - Chen Luo
- Department of General Surgery & Pancreatic Injury and Repair
Key Laboratory of Sichuan Province, The General Hospital of Western Theater
Command, Chengdu 610083, China
- Department of Hepatopancreatobiliary Surgery, Panzhihua Central
Hospital, Panzhihua 617000, China
| | - Jun Wu
- College of Medicine, Southwest Jiaotong University, Chengdu
610031, China
- Department of General Surgery & Pancreatic Injury and Repair
Key Laboratory of Sichuan Province, The General Hospital of Western Theater
Command, Chengdu 610083, China
| | - Wei Li
- Laboratory of Basic Medical Sciences, The General Hospital of
Western Theater Command, Chengdu 610083, China
| | - Xin Guo
- Laboratory of Basic Medical Sciences, The General Hospital of
Western Theater Command, Chengdu 610083, China
| | - Shuai Li
- College of Medicine, Southwest Jiaotong University, Chengdu
610031, China
- Department of General Surgery & Pancreatic Injury and Repair
Key Laboratory of Sichuan Province, The General Hospital of Western Theater
Command, Chengdu 610083, China
| | - Bing Wang
- College of Medicine, Southwest Jiaotong University, Chengdu
610031, China
- Department of General Surgery & Pancreatic Injury and Repair
Key Laboratory of Sichuan Province, The General Hospital of Western Theater
Command, Chengdu 610083, China
| | - Hongyu Sun
- College of Medicine, Southwest Jiaotong University, Chengdu
610031, China
- Department of General Surgery & Pancreatic Injury and Repair
Key Laboratory of Sichuan Province, The General Hospital of Western Theater
Command, Chengdu 610083, China
- Laboratory of Basic Medical Sciences, The General Hospital of
Western Theater Command, Chengdu 610083, China
| | - Lijun Tang
- College of Medicine, Southwest Jiaotong University, Chengdu
610031, China
- Department of General Surgery & Pancreatic Injury and Repair
Key Laboratory of Sichuan Province, The General Hospital of Western Theater
Command, Chengdu 610083, China
| |
Collapse
|
|
3
|
Abdominal paracentesis drainage attenuates severe acute pancreatitis by enhancing cell apoptosis via PI3K/AKT signaling pathway. Apoptosis 2021; 25:290-303. [PMID: 32100210 PMCID: PMC7181427 DOI: 10.1007/s10495-020-01597-2] [Citation(s) in RCA: 20] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
Our previous studies have shown that abdominal paracentesis drainage (APD) is a safe and effective strategy for patients with severe acute pancreatitis (SAP). However, the underlying mechanisms behind APD treatment remain poorly understood. Given that apoptosis is a critical pathological response of SAP, we here aim to investigate the effect of APD on cell apoptosis in pancreatic tissues during SAP and to explore its potential molecular mechanism. SAP was induced by 5% sodium-taurocholate retrograde while APD group was inserted a drainage tube into the right lower abdomen of rats immediately after SAP induction. Histopathological staining, serum amylase, endotoxin and inflammatory mediators were measured. Cell apoptosis, apoptosis-related proteins and signaling pathway were also evaluated. Our results demonstrated that APD treatment significantly attenuated pancreatic damage and decreased the serum levels of amylase, endotoxin, TNF-α, IL-1 and IL-6 in rats with SAP. Notably, APD treatment enhanced cell apoptosis and reduced necrosis in pancreatic tissues, as evidenced by Tunnel staining, the increased pro-apoptosis proteins (Cleaved-caspase-3 and bax) and decreased anti-apoptosis protein (Bcl-2). Moreover, the effect of APD on cell apoptosis was further confirmed by the regulatory pathway of PI3K/AKT and NF-kB signaling pathway. These results suggest that APD attenuates the severity of SAP by enhancing cell apoptosis via suppressing PI3K/AKT signaling pathway. Our findings provide new insights for understanding the effectiveness of APD in patients with SAP.
Collapse
|
|
4
|
Zhao R, Song C, Liu L, Liu Q, Zhou N, Zhou X, Xie Y. Single immunoglobulin and Toll‑interleukin‑1 receptor domain containing molecule protects against severe acute pancreatitis in vitro by negatively regulating the Toll‑like receptor‑4 signaling pathway: A clinical and experimental study. Mol Med Rep 2020; 22:2851-2859. [PMID: 32945488 PMCID: PMC7453662 DOI: 10.3892/mmr.2020.11379] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/13/2020] [Accepted: 07/10/2020] [Indexed: 02/05/2023] Open
Abstract
Single immunoglobulin and Toll-interleukin-1 receptor domain-containing molecule (SIGIRR) is a specific inhibitor of IL-1R and Toll-like receptor (TLR) signaling and considered a potential target for the treatment of inflammatory diseases. Pathogenic mechanisms associated with the TLR4 signaling pathway have a critical role in the development of severe acute pancreatitis (SAP). The aim of the present study was to determine the role of SIGIRR in the regulation of TLR4 signaling during the progression of SAP. Pancreatitis-associated ascitic fluid (PAAF) was collected from patients with SAP. Murine RAW264.7 macrophages were transfected with a SIGIRR overexpression plasmid and co-cultured with the PAAF from the donors in order to evaluate the effect of SIGIRR in vitro. The mRNA expression of TLR4, SIGIRR and other key downstream signaling molecules was quantified using semi-quantitative PCR with agarose gel electrophoresis. Furthermore, the levels of pro-inflammatory cytokines in the culture supernatant were detected using ELISA. In contrast to SIGIRR, the mRNA expression levels of TLR4, myeloid differentiation factor 88 (MyD88), IL-1R-associated kinase-1 (IRAK-1) and TNF receptor-associated factor-6 (TRAF-6) were significantly increased in RAW264.7 cells following treatment with PAAF. Furthermore, TLR4, MyD88, IRAK-1 and TRAF-6 mRNA levels were significantly downregulated following SIGIRR overexpression and PAAF treatment in RAW264.7 cells. The levels of IL-2, IL-12, IL-17 and IFN-γ in the culture supernatant were also significantly decreased, while IL-10 levels were increased. Overall, SIGIRR negatively regulated the TLR4 signaling pathway to protect against the development of SAP in an in vitro model. Therefore, SIGIRR may represent a promising therapeutic target for SAP.
Collapse
Affiliation(s)
- Rulin Zhao
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Conghua Song
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Li Liu
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Qiong Liu
- Jiangxi Institute of Medical Sciences, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Nanjin Zhou
- Jiangxi Institute of Medical Sciences, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Xiaojiang Zhou
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Yong Xie
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| |
Collapse
|
|
5
|
Samanta J, Rana A, Dhaka N, Agarwala R, Gupta P, Sinha SK, Gupta V, Yadav TD, Kochhar R. Ascites in acute pancreatitis: not a silent bystander. Pancreatology 2019; 19:646-652. [PMID: 31301995 DOI: 10.1016/j.pan.2019.06.004] [Citation(s) in RCA: 24] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/10/2019] [Revised: 05/03/2019] [Accepted: 06/08/2019] [Indexed: 12/11/2022]
Abstract
BACKGROUND & AIM Ascites in patients with acute pancreatitis (AP) is understudied although recent literature hints at its evident role in the final outcome. This study was planned to study the characteristics of ascites in patients of AP and its effect on the disease course and outcome. METHODS Consecutive patients of AP were studied and patients with or without ascites were evaluated for the baseline parameters and severity assessment. Ascites was quantified and fluid analyzed for its characteristics. Intraabdominal pressure (IAP) was monitored. The various outcome parameters were compared between the two groups of patients with and without ascites. RESULTS Of the cohort of 213 patients, 82 (38.5%) developed ascites. Ascites group had significantly higher rates of organ failure (p = 0.001), necrosis (p=<0.001) and higher severity assessment scores. The ascites group had significantly longer hospital and ICU stay and higher ventilator days compared to the non-ascites group. Mortality was also higher in the ascites group (34.1% vs 8.45; p = 0.001). Majority of patients with ascites had moderate to gross ascites (75.6%), low serum ascites albumin gradient (87.8%) with low amylase levels (71.9%). Sub-group analysis in ascites group showed that patients with fatal outcome had higher rates of moderate to gross ascites, higher baseline IAP and lower reduction in IAP after 48 h. Moderate to gross ascites and grades of intra-abdominal hypertension (IAH) were significant predictors of mortality (AUC - 0.76). CONCLUSION AP patients with ascites have a more severe disease with poorer outcome. Higher degrees of ascites and IAH grades are significant predictors of mortality.
Collapse
Affiliation(s)
- Jayanta Samanta
- Department of Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Atul Rana
- Department of Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Narendra Dhaka
- Department of Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Roshan Agarwala
- Department of Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Pankaj Gupta
- Department of Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Saroj Kant Sinha
- Department of Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Vikas Gupta
- Department of Surgery, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Thakur Deen Yadav
- Department of Surgery, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Rakesh Kochhar
- Department of Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
| |
Collapse
|
|
6
|
Liu RH, Wen Y, Sun HY, Liu CY, Zhang YF, Yang Y, Huang QL, Tang JJ, Huang CC, Tang LJ. Abdominal paracentesis drainage ameliorates severe acute pancreatitis in rats by regulating the polarization of peritoneal macrophages. World J Gastroenterol 2018; 24:5131-5143. [PMID: 30568390 PMCID: PMC6288649 DOI: 10.3748/wjg.v24.i45.5131] [Citation(s) in RCA: 23] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/26/2018] [Revised: 10/20/2018] [Accepted: 11/09/2018] [Indexed: 02/06/2023] Open
Abstract
AIM To investigate the role of peritoneal macrophage (PM) polarization in the therapeutic effect of abdominal paracentesis drainage (APD) on severe acute pancreatitis (SAP).
METHODS SAP was induced by 5% Na-taurocholate retrograde injection in Sprague-Dawley rats. APD was performed by inserting a drainage tube with a vacuum ball into the lower right abdomen of the rats immediately after the induction of SAP. To verify the effect of APD on macrophages, PMs were isolated and cultured in an environment, with the peritoneal inflammatory environment simulated by the addition of peritoneal lavage in complete RPMI 1640 medium. Hematoxylin and eosin staining was performed. The levels of pancreatitis biomarkers amylase and lipase as well as the levels of inflammatory mediators in the blood and peritoneal lavage were determined. The polarization phenotypes of the PMs were identified by detecting the marker expression of M1/M2 macrophages via flow cytometry, qPCR and immunohistochemical staining. The protein expression in macrophages that had infiltrated the pancreas was determined by Western blot.
RESULTS APD treatment significantly reduced the histopathological scores and levels of amylase, lipase, tumor necrosis factor-α and interleukin (IL)-1β, indicating that APD ameliorates the severity of SAP. Importantly, we found that APD treatment polarized PMs towards the M2 phenotype, as evidenced by the reduced number of M1 macrophages and the reduced levels of pro-inflammatory mediators, such as IL-1β and L-selectin, as well as the increased number of M2 macrophages and increased levels of anti-inflammatory mediators, such as IL-4 and IL-10. Furthermore, in an in vitro study wherein peritoneal lavage from the APD group was added to the cultured PMs to simulate the peritoneal inflammatory environment, PMs also exhibited a dominant M2 phenotype, resulting in a significantly lower level of inflammation. Finally, APD treatment increased the proportion of M2 macrophages and upregulated the expression of the anti-inflammatory protein Arg-1 in the pancreas of SAP model rats.
CONCLUSION These findings suggest that APD treatment exerts anti-inflammatory effects by regulating the M2 polarization of PMs, providing novel insights into the mechanism underlying its therapeutic effect.
Collapse
Affiliation(s)
- Ruo-Hong Liu
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
- Third Military Medical University (Army Medical University), Chongqing 400037, China
| | - Yi Wen
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
- Third Military Medical University (Army Medical University), Chongqing 400037, China
| | - Hong-Yu Sun
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
| | - Chun-Yu Liu
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
| | - Yu-Fan Zhang
- Jiaotong Hospital Affiliated with the Sichuan Provincial People’s Hospital, Chengdu 611730, Sichuan Province, China
| | - Yi Yang
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
| | - Qi-Lin Huang
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
| | - Jia-Jia Tang
- Department of Ultrasound, Chinese Academy of Medical Sciences and Peking Union Medical College Hospital, Beijing 100032, China
| | - Can-Chen Huang
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
| | - Li-Jun Tang
- PLA Center of General Surgery and Pancreatic Injury and Repair Key Laboratory of Sichuan Province, Chengdu Military General Hospital, Chengdu 610083, Sichuan Province, China
- Third Military Medical University (Army Medical University), Chongqing 400037, China
| |
Collapse
|
|
7
|
Wang J, Zhang C, Xu P, Yang ZW, Weng CZ, Lai YX. Phosphoinositide 3‑kinase/protein kinase B regulates inflammation severity via signaling of Toll‑like receptor 4 in severe acute pancreatitis. Mol Med Rep 2018; 17:7835-7844. [PMID: 29620213 DOI: 10.3892/mmr.2018.8819] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/26/2017] [Accepted: 03/21/2018] [Indexed: 11/05/2022] Open
Abstract
Phosphatidylinositol 3‑kinase (PI3K)/protein kinase B (Akt) has been indicated to serve an important role in the pathogenesis of inflammatory diseases. It was previously demonstrated that the PI3K/Akt inhibitor wortmannin alleviated the severity of inflammation and improved the survival rate in rats with induced severe acute pancreatitis (SAP), which indicates that PI3K/Akt may serve a role in the pathogenesis of acute pancreatitis. To date, the mechanism by which PI3K/Akt regulates inflammation has not been elucidated. In the present study, it was hypothesized that PI3K/Akt may be invovled in SAP inflammation via regulation of the Toll‑like receptor 4 (TLR4) signaling pathway. Rats with SAP were treated with the PI3K/Akt agonist insulin‑like growth factor (IGF)‑1, which alleviated the severity of inflammation in a dose‑dependent manner. Furthermore, to better understand the role of PI3K/Akt in inflammation, RAW264.7 murine macrophages were stimulated with IGF‑1 and wortmannin alone or together before the induction of inflammation by treatment with lipopolysaccharide (LPS). The results indicated that LPS stimulated overexpression of TLR4, myeloid differentiation primary response gene 88 (MyD88), PI3K, Akt, p38MAPK and NF‑κBp65 mRNA, and increased the levels of tumor necrosis factor (TNF)‑α and interleukin (IL)‑6 in RAW264.7 cells compared with the control group. The levels of all detected factors were increased by stimulation with IGF‑1, whereas these levels were decreased following treatment with wortmannin alone, and the effect of IGF‑1 was abolished by wortmannin in RAW264.7 cells. In vivo studies indicated that IGF‑1 produced the same anti‑inflammatory effect as wortmannin and that expression of TLR4, p38MAPK and NF‑κBp65 decreased following treatment with IGF‑1. These findings indicate that PI3K/Akt may take part in the progression of SAP by regulating the TLR4 signaling pathway and that IGF‑1 can inhibit inflammation in SAP rats.
Collapse
Affiliation(s)
- Jing Wang
- Department of Gastroenterology, Shanghai Songjiang Hospital Affiliated to Nanjing Medical University, Shanghai 201600, P.R. China
| | - Chun Zhang
- Department of Gastroenterology, Shanghai Songjiang Hospital Affiliated to Nanjing Medical University, Shanghai 201600, P.R. China
| | - Ping Xu
- Department of Gastroenterology, Shanghai Songjiang Hospital Affiliated to Nanjing Medical University, Shanghai 201600, P.R. China
| | - Zhi-Wen Yang
- Department of Pharmacy, Songjiang Hospital Affiliated Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 201600, P.R. China
| | - Cheng-Zhao Weng
- Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200030, P.R. China
| | - Yue-Xing Lai
- Department of Gastroenterology, Shanghai Songjiang Hospital Affiliated to Nanjing Medical University, Shanghai 201600, P.R. China
| |
Collapse
|
|
8
|
Bacillus subtilis CFR5 isolated from fermented soybean attenuates the chronic pancreatitis. J Funct Foods 2018. [DOI: 10.1016/j.jff.2017.11.009] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022] Open
|
|
9
|
Sharmila GR, Venkateswaran G. Protective effect of bacillopeptidase CFR5 from Bacillus subtilis CFR5 on cerulein-induced pancreatitis. Biochem Biophys Res Commun 2017; 491:455-462. [PMID: 28709869 DOI: 10.1016/j.bbrc.2017.07.054] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/02/2017] [Accepted: 07/10/2017] [Indexed: 01/05/2023]
Abstract
Bacillopeptidase is a serine peptidase, known for its fibrinolytic activity. However, a very little information is known about its in vivo inflammatory and/or anti-inflammatory properties. Thus, to understand whether bacillopeptidase incorporation can regulate pancreatitis or not, the cerulein-induced pancreatitis model was used, and the role of bacillopeptidase on pancreatitis was studied. In this study, 46 kDa protein was purified from Bacillus subtilis and identified as bacillopeptidase CFR5 (BPC) through MS/MS analysis. The nutritional prophylactic group was orally fed with two doses of BPC (100 μg/Kg/BW of rat) 6 h before cerulein administration and analyzed for its effect on intestine and pancreas inflammation, cytokines, and pancreatitis marker gene expression. BPC administration significantly reduced the severity of pancreatitis by decreasing serum amylase, lipase, pancreatic edema and myeloperoxidase activity. The pretreatment with BPC suppressed the pancreatic pro-inflammatory and inflammatory cytokines production including IL-6, IL-1β, TNF-α, IL-2, IL-4, IL-5, IL-10, and IL-13 in both pancreas and serum samples. Moreover, BPC supplementation restored pancreatitis mediated disruption of intestinal barrier integrity by upregulating tight junction proteins (ZO-1, occludin), antimicrobial peptides (DEFB1, CRAMP), MUC-2, TFF3 expression and by enhancing SCFA's production. Pretreatment with BPC suppressed the intestinal inflammation with reduced cytokines production in the colon and ileal region of cerulein-induced pancreatitis. Thus, BPC based pretreatment protocol is a novel intervention to prevent acute pancreatitis.
Collapse
Affiliation(s)
- G R Sharmila
- Academy of Scientific and Innovative Research, CSIR-Central Food Technological Research Institute, Mysuru 570020, India; Microbiology and Fermentation Technology Department, CSIR-Central Food Technological Research Institute, Mysuru 570020, India
| | - G Venkateswaran
- Academy of Scientific and Innovative Research, CSIR-Central Food Technological Research Institute, Mysuru 570020, India; Microbiology and Fermentation Technology Department, CSIR-Central Food Technological Research Institute, Mysuru 570020, India.
| |
Collapse
|
|
10
|
Xiang H, Wang G, Qu J, Xia S, Tao X, Qi B, Zhang Q, Shang D. Yin-Chen-Hao Tang Attenuates Severe Acute Pancreatitis in Rat: An Experimental Verification of In silico Network Target Prediction. Front Pharmacol 2016; 7:378. [PMID: 27790147 PMCID: PMC5061810 DOI: 10.3389/fphar.2016.00378] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/08/2016] [Accepted: 09/28/2016] [Indexed: 12/12/2022] Open
Abstract
Yin-Chen-Hao Tang (YCHT) is a classical Chinese medicine compound that has a long history of clinical use in China for the treatment of inflammatory diseases. However, the efficacy and mechanisms of YCHT for the treatment of severe acute pancreatitis (SAP) are not known. The current study investigated the pharmacological properties of YCHT against SAP and its underlying mechanisms. A computational prediction of potential targets of YCHT was initially established based on a network pharmacology simulation. The model suggested that YCHT attenuated SAP progress by apoptosis inducement, anti-inflammation, anti-oxidation and blood lipid regulation. These effects were validated in SAP rats. YCHT administration produced the following results: (1) significantly inhibited the secretion of pancreatic enzymes and protected pancreatic tissue; (2) obviously increased the number of in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL)-positive cells and induced apoptosis; (3) markedly inhibited neutrophil infiltration to the impaired pancreas and reduced the inflammatory reaction; (4) notably enhanced the activities of antioxidant enzymes and decreased the nitric oxide synthase levels; (5) significantly reduced the levels of triglycerides, total cholesterol and low-density lipoprotein and increased high-density lipoprotein; and (6) significantly up-regulated peroxisome proliferator-activated receptor-γ (PPARγ) and down-regulated nuclear factor-kappa B (NF-κB). In summary, these results demonstrated that YCHT attenuated SAP progress by inducing apoptosis, repressing inflammation, alleviating oxidative stress and regulating lipid metabolism partially via regulation of the NF-κB/PPARγ signal pathway.
Collapse
Affiliation(s)
- Hong Xiang
- College (Institute) of Integrative Medicine, Dalian Medical UniversityDalian, China
| | - Guijun Wang
- Department of General Surgery, The First Affiliated Hospital of Jinzhou Medical UniversityJinzhou, China
| | - Jialin Qu
- Clinical Laboratory of Integrative Medicine, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
| | - Shilin Xia
- Clinical Laboratory of Integrative Medicine, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
| | - Xufeng Tao
- College of Pharmacy, Dalian Medical UniversityDalian, China
| | - Bing Qi
- Department of General Surgery, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
| | - Qingkai Zhang
- Department of General Surgery, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
| | - Dong Shang
- College (Institute) of Integrative Medicine, Dalian Medical UniversityDalian, China
- Department of General Surgery, The First Affiliated Hospital of Dalian Medical UniversityDalian, China
| |
Collapse
|
|
11
|
Abstract
OBJECTIVE The aim of this study was to assess the efficiency and safety of peritoneal lavage in patients with severe acute pancreatitis. METHODS A comprehensive search was performed to identify randomized controlled trials that compared peritoneal lavage with conservative treatment for severe acute pancreatitis. The primary outcome was all-cause mortality. Secondary outcomes included complications rate, proportion of need for operation, length of hospital stay, and medical costs. RESULTS A total of 899 patients from 15 studies were subjected to this systematic review. Peritoneal lavage did significantly decrease the mortality (relative risk, 0.47; 95% confidence interval, 0.34-0.66; P < 0.01), with low heterogeneity among the studies (I = 7%). Peritoneal lavage also seemed to significantly alter any of the other end points. CONCLUSIONS Peritoneal lavage shows robustly beneficial effect in patients with severe acute pancreatitis in this systematic review. However, the results should be interpreted with caution due to the general high risk of bias in these included studies.
Collapse
|
|
12
|
Chen P, Wang W, Zhang Y, Yuan Y, Wu Y. Decreased MIZ1 Expression in Severe Experimental Acute Pancreatitis: A Rat Study. Dig Dis Sci 2016; 61:758-66. [PMID: 26581215 DOI: 10.1007/s10620-015-3951-y] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/25/2015] [Accepted: 10/27/2015] [Indexed: 12/15/2022]
Abstract
AIM We tested our hypothesis that Myc-interacting zinc finger protein 1 (MIZ1), a cell cycle regulator, suppressed inflammation, and therefore, represented a useful prognostic marker in patients with acute necrotizing pancreatitis (ANP) complicated by acute lung injury. METHODS Sprague-Dawley rats were randomly divided into control and ANP groups at different time points. The MIZ1 protein expression was measured by Western blot and ELISA, and confirmed using immunohistochemistry. The severity of pancreatic and lung injury was evaluated by the injury score and wet/dry weight ratio. The severity of disease was evaluated by serum C-reactive protein (CRP). The MPO activity of lung tissue amylase levels and the degree of inflammation were evaluated by serum tumor necrosis factor (TNF)-α and interleukin (IL)-6 expression. The risk due to multiple factors was investigated by relationship analysis. RESULTS The serum levels of CRP, amylase, TNF-α, and IL-6 were gradually increased at 6, 24, and 48 h in ANP when compared with the control rats. The MIZ1 expressions were greatly decreased in ANP rats, especially at 24 h. Statistical analysis showed that there were time-dependent differences in ANP rats when compared with control rats (6 vs. 24 or 48 h, P < 0.01). MIZ1 showed close negative correlation with the degree of pancreatic and lung injury, serum amylase, CRP, TNF-α, and IL-6 (P < 0.01, respectively). CONCLUSION The decreasing MIZ1 expression was closely correlated with inflammatory response, and development of ANP. Decreasing MIZ1 levels indicate a risk for ANP.
Collapse
Affiliation(s)
- Ping Chen
- Department of Gastroenterology, Ruijin Hospital North, Shanghai Jiaotong University School of Medicine, Shanghai, 201801, China.
| | - Weiyi Wang
- Department of Gastroenterology, Ruijin Hospital North, Shanghai Jiaotong University School of Medicine, Shanghai, 201801, China.
| | - Yongping Zhang
- Department of Gastroenterology, Ruijin Hospital North, Shanghai Jiaotong University School of Medicine, Shanghai, 201801, China.
| | - Yaozong Yuan
- Department of Gastroenterology, Ruijin Hospital North, Shanghai Jiaotong University School of Medicine, Shanghai, 201801, China.
| | - Yunlin Wu
- Department of Gastroenterology, Ruijin Hospital North, Shanghai Jiaotong University School of Medicine, Shanghai, 201801, China.
| |
Collapse
|
|
13
|
Yang ZW, Weng CZ, Wang J, Xu P. The role of Card9 overexpression in peripheral blood mononuclear cells from patients with aseptic acute pancreatitis. J Cell Mol Med 2015; 20:441-9. [PMID: 26893103 PMCID: PMC4759462 DOI: 10.1111/jcmm.12738] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2015] [Accepted: 10/18/2015] [Indexed: 01/04/2023] Open
Abstract
Activated mononuclear cells are an early event in the course of severe acute pancreatitis (SAP). To date, the molecular mechanism triggering peripheral blood mononuclear cells (PBMCs) is poorly understood. The aim of this paper was to determine the potential role of Card9 in SAP. We collected data from 72 subjects between January 2013 and June 2014. Subsequently, PBMCs were isolated on day 1, 3 and 5 of pancreatitis. Immunofluorescence staining, quantitative real‐time PCR, Western blotting, immunoprecipitation and ELISA were used to determine the role of Card9 in SAP. Microbial culture showed that SAP patients at the early period did not develop any bacteria and fungi infection. Card9 expression in SAP patients was higher than that in mild acute pancreatitis and volunteer healthy controls, up to the peak on day 1. The monocyte‐derived cytokines interleukin (IL)‐17, IL‐1β, IL‐6 and tumour necrosis factor‐α mediated by the induction of Card9 markedly increased in SAP patients compared with the control group. Furthermore, the inducible formation of Card9‐Bcl10 complex was found in PBMCs, which may be involved in nuclear factor kappa B (NF‐κB) and p38 activation in SAP. Receiver operating characteristic curve indicated that Card9 levels had a high sensitivity of 87.5% and specificity of 67.7%, showing the close correlation with SAP patients. Card9 overexpression was firstly found in aseptic SAP, which may be played an important role in NF‐κB and p38 activation in PBMCs. It also provided the new insights into therapeutic interventions by targeting monocytes activation in SAP patients.
Collapse
Affiliation(s)
- Zhi-wen Yang
- Department of Pharmacy, Songjiang Hospital Affiliated Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai, China
| | - Cheng-zhao Weng
- Department of Digestive, Shanghai Songjiang Hospital Affiliated to Nanjing Medical University, Nanjing, China
| | - Jing Wang
- Department of Digestive, Songjiang Hospital Affiliated Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai, China
| | - Ping Xu
- Department of Digestive, Shanghai Songjiang Hospital Affiliated to Nanjing Medical University, Nanjing, China.,Department of Digestive, Songjiang Hospital Affiliated Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai, China
| |
Collapse
|
|
14
|
Zhao HX, Yang XH, Li CP, Chen X. Small intestinal smooth muscle cell apoptosis in rats with severe acute pancreatitis. Shijie Huaren Xiaohua Zazhi 2014; 22:4231-4236. [DOI: 10.11569/wcjd.v22.i28.4231] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate whether small intestinal smooth muscle cell apoptosis occurs in rats with severe acute pancreatitis (SAP) and the underlying mechanism.
METHODS: Male SD rats were randomly divided into a sham operation group (SO) and an SAP group. SAP was induced by injecting 3.8% sodium taurocholate solution into the subcapsular region of the pancreas of SD rats. Rats in the SO group were injected with 1 mL/kg normal saline. Forty-eight hours later, pancreatic pathological changes and the transit rate of the small bowel were determined. Cell apoptosis, expression of adenine nucleotide translocator (ANT) mRNA, mitochondrial membrane potential, and cytochrome C (Cyt-C) protein expression in the small intestinal smooth muscle were determined by TUNEL method, RT-PCR, flow cytometry and immunohistochemistry, respectively.
RESULTS: Compared with rats in the SO group, rats in the SAP group developed typical SAP symptoms, with a higher pancreatic pathology score (6.85 ± 1.21 vs 1.13 ± 0.91, P < 0.001). Compared with rats in the SO group, the transit rate of the small intestine was significantly lower (55.91% ± 2.93% vs 68.9% ± 5.69%, P < 0.05), the apoptosis of smooth muscle cells in the small intestine increased significantly (0.056 ± 0.184 vs 0.029 ± 0.038, P < 0.05), the expression of ANT mRNA and Cyt-C protein (0.024 ± 0.001 vs 0.057 ± 0.168, P < 0.001) in the smooth muscle of the small intestine increased significantly, and the mitochondrial membrane potential decreased significantly (5.07 ± 0.92 vs 2.40 ± 0.50, P < 0.05) in the SAP group.
CONCLUSION: The mitochondrial signal transduction pathway contributes to smooth muscle cell apoptosis in the small intestine, which may play a role in small intestinal motility dysfunction in SAP rats.
Collapse
|
|
15
|
Chen SM, Tsai YS, Lee SW, Liu YH, Liao SK, Chang WW, Tsai PJ. Astragalus membranaceus modulates Th1/2 immune balance and activates PPARγ in a murine asthma model. Biochem Cell Biol 2014; 92:397-405. [PMID: 25264079 DOI: 10.1139/bcb-2014-0008] [Citation(s) in RCA: 32] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022] Open
Abstract
Astragalus membranaceus, a traditional Chinese herb, has been used to improve airway inflammation and asthma. The present study investigated whether A. membranaceus has immunotherapeutic effects on asthma, a chronic inflammatory mucosal disease that is associated with excess production of IgE, eosinophilia, T helper 2 (Th2) cytokines, and bronchial hyperresponsiveness. An ovalbumin (OVA)-induced, chronic inflammatory airway murine asthma model was used to examine the status of pulmonary inflammation after the administration of A. membranaceus. The IgE levels in serum and bronchoalveolar lavage fluid showed a tendency to decrease after the administration of A. membranaceus. The number of eosinophils decreased and infiltration of inflammatory cells and collagen deposition declined in lung sections after A. membranaceus administration. The RNA and protein levels of Th2 cytokines and the ratio of the GATA3/T-bet mRNA levels decreased after A. membranaceus treatment. Furthermore, the mRNA level of peroxisome proliferator-activated receptor γ (PPARγ), a nuclear hormone receptor, increased in the lung tissues of A. membranaceus-treated mice. Finally, an A. membranaceus water extract activated PPARγ activity in either human embryonic kidney 293 (HEK293) or A549 cells in a PPARγ-responsive element-containing luciferase reporter assay. These results indicate that A. membranaceus has an inhibitory effect on airway inflammation in a murine model of asthma through modulating the imbalanced relationship between Th1 and Th2 cytokines.
Collapse
Affiliation(s)
- Shih-Ming Chen
- a Graduate Institute of Clinical Medical Sciences, Chang Gung University, Tao-Yuan, Taiwan
| | | | | | | | | | | | | |
Collapse
|
|
16
|
Liu LL, Wang XY. Severe acute pancreatitis complicated with gastrointestinal dysfunction: Pathogenesis, diagnosis and treatment. Shijie Huaren Xiaohua Zazhi 2013; 21:3828-3834. [DOI: 10.11569/wcjd.v21.i34.3828] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
Severe acute pancreatitis (SAP) is often associated with gastrointestinal dysfunction, leading to gastrointestinal motility disorders and even gastrointestinal failure, which has an important effect on SAP progression and prognosis, directly influences the outcome of treatment, is an important cause of death in patients with SAP, and moreover, has been one of the important prognostic factors for SAP. This review aims to discuss the pathophysiology, pathogenesis, diagnosis and treatment of SAP with gastrointestinal dysfunction.
Collapse
|
|
17
|
Xu P, Lou XL, Chen C, Yang ZW. Effects of peroxisome proliferator-activated receptor-γ activation on apoptosis in rats with acute pancreatitis. Dig Dis Sci 2013; 58:3516-23. [PMID: 24185678 DOI: 10.1007/s10620-013-2842-3] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/05/2013] [Accepted: 08/08/2013] [Indexed: 01/12/2023]
Abstract
PURPOSE To investigate the effects and mechanisms of peroxisome proliferator-activated receptor-γ (PPAR-γ) activation on the induction of apoptosis in rats with acute pancreatitis. METHODS Severe acute pancreatitis (SAP) and mild acute pancreatitis (MAP) were induced and pre-treated with pioglitazone, which is a ligand of PPAR-γ. The expression of inflammatory factors (TNF-α and IL6) of the pancreas was detected by ELISA. The apoptosis in pancreas were detected by TUNEL assay and the activity of caspase 3 was determined. Phosphorylation of p65 in pancreas of SAP or MAP was determined by western-blot. RESULTS Expression levels of PPAR-γ proteins were elevated in the pancreases of SAP or MAP rats pre-injected with pioglitazone intraperitoneally. Downregulation of the expression TNF-α and IL6 and relief of pathological changes in the pancreas suggested that pioglitazone had protective effects on acute panceatitis. In pioglitazone pre-treated groups, a TUNEL assay indicated a high level of apoptosis in SAP but little apoptosis in MAP, showing pioglitazone could promote taurocholate-induced apoptosis but inhibit ceruleininduced apoptosis in pancraeatic aniniar cells. Furthermore, caspase 3 activity was high in SAP but low in MAP, implying that the apoptotic mechanism in pancreatic acinar cells of AP rats was correlated with caspase 3 activity. Phosphorylation of p65 was reduced in SAP or MAP group pretreated with pioglitazone, indicating that pioglitazone reduced the inflammation reaction by inhibiting the activation of the NF-κB. CONCLUSIONS These results indicated that activation of PPAR-γ induced apoptosis in pancreatic acinar cells of SAP rats but inhibited apoptosis in pancraeatic acinar cells of MAP rats, which demonstrated that PPAR-γ may be an efficiently therapeutic target in pancreatic inflammation.
Collapse
Affiliation(s)
- Ping Xu
- Department of Gastroenterology, Songjiang Hospital Affiliated First People's Hospital, Shanghai Jiao Tong University, Shanghai, 201600, China,
| | | | | | | |
Collapse
|