|Year : 2019 | Volume
| Issue : 6 | Page : 763-770
Association of endotoxaemia & gut permeability with complications of acute pancreatitis: Secondary analysis of data
Namrata Singh1, Ujjwal Sonika1, Praneeth Moka1, Brij Sharma1, Vikas Sachdev1, Sushil Kumar Mishra1, Ashish Datt Upadhyay2, Anoop Saraya1
1 Department of Gastroenterology, All India Institute of Medical Sciences, New Delhi, India
2 Department of Biostatistics, All India Institute of Medical Sciences, New Delhi, India
|Date of Submission||11-May-2017|
|Date of Web Publication||3-Sep-2019|
Dr Anoop Saraya
Department of Gastroenterology & Human Nutrition Unit, Teaching Block, All India Institute of Medical Sciences, New Delhi 110 029
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background & objectives: In acute pancreatitis (AP) gut barrier dysfunction is considered as an important predisposing factor leading to increased intestinal permeability (IP). In this study a pooled analysis of data published in our previous four studies on various aspects of gut permeability and endotoxaemia in patients with AP was attempted to find an association between increased IP and severity of disease and associated complications.
Methods: This study was a pooled analysis of data of four previously published prospective studies on AP. Gut permeability, assessed by lactulose/mannitol excretion in urine and endotoxin core antibodies type IgG and IgM (EndoCab IgG and IgM) were measured on days zero and seven (D0 and D7) of admission. All patients received standard treatment of AP. We studied whether IgG and IgM anti-endotoxin titres and lactulose-mannitol ratio (LMR) at admission and D7 were associated with organ failure, infection and mortality.
Results: The titres of anti-endotoxin IgG and IgM were lower in all patients of AP (n=204), both in mild AP (n=24) and severe AP (n=180) in the first week, compared to controls (n=15). There was no significant difference in serum IgG and IgM anti-endotoxin levels and LMR at baseline and at D7 among patients with organ failure, infection and mortality.
Interpretation & conclusions: Our findings showed that serum IgG and IgM anti-endotoxin titres and LMR at admission and at day 7 were not associated with organ failure, infection, and death of patients with AP.
Keywords: Acute pancreatitis - endotoxaemia - intestinal permeability - mortality - organ failure
|How to cite this article:|
Singh N, Sonika U, Moka P, Sharma B, Sachdev V, Mishra SK, Upadhyay AD, Saraya A. Association of endotoxaemia & gut permeability with complications of acute pancreatitis: Secondary analysis of data. Indian J Med Res 2019;149:763-70
|How to cite this URL:|
Singh N, Sonika U, Moka P, Sharma B, Sachdev V, Mishra SK, Upadhyay AD, Saraya A. Association of endotoxaemia & gut permeability with complications of acute pancreatitis: Secondary analysis of data. Indian J Med Res [serial online] 2019 [cited 2020 Feb 22];149:763-70. Available from: http://www.ijmr.org.in/text.asp?2019/149/6/763/265964
In acute pancreatitis (AP), mortality during late phase occurs due to the development of severe pancreatic and peripancreatic infection leading to sepsis and multiorgan dysfunction. An important predisposing factor for infection is gut barrier dysfunction. Both experimental and human studies have demonstrated increased intestinal permeability (IP) in severe AP,,,,,. AP-induced hypovolaemia due to endothelial barrier leakage and gut arteriovenous shunting causes intestinal ischaemia and reperfusion injury with concomitant gut barrier dysfunction. Increase in IP results in translocation of Gram-negative bacteria (GNB) through the lymphatics via the mesenteric nodes,. An antibody response is stimulated against GNB endotoxins attached to the cell wall. Some studies have noted that endotoxaemia is associated with systemic inflammatory response syndrome (SIRS), multiorgan failure and high mortality,.
In our previous study on the role of probiotics on gut permeability and endotoxaemia in patients of AP, no significant trend was identified for an effect of probiotics on gut permeability or endotoxaemia in AP. However, this study was underpowered owing to premature study termination. In another study to determine the non-inferiority of early enteral feeding through nasogastric (NG) compared to nasojejunal (NJ) route on infectious complications in patients with severe AP, infectious complications were found to be within the non-inferiority limit. Pain in refeeding, IP and endotoxaemia were comparable in both groups. We published four studies,,, on AP during 2011 to 2014 which looked into IP and endotoxaemia in patients with AP. In the present study, a pooled analysis of data published in these four studies,,, was attempted with the hypothesis that the increased IP leads to severe disease in patients with AP along with increased complications and mortality.
| Material & Methods|| |
This study was a secondary pooled data analysis of our four previously conducted prospective studies,,, between June 2006 and December 2011. Patients of AP admitted in the department of Gastroenterology and Human Nutrition Unit of the All India Institute of Medical Sciences (AIIMS), New Delhi, India, during these studies were included for data analysis. Patients who presented with proven or suspected infected pancreatic necrosis and patients who had been put on enteral nutrition before admission were excluded from the analysis.
AP was diagnosed in the presence of at least two of the following: (i) Presence of pancreatic pain; (ii) at least three-fold rise in serum amylase; and (iii) evidence of AP on imaging studies.
The amount of pancreatic necrosis was graded as <30, 30-50 and >50 per cent. Computed tomography severity index (CTSI) was calculated as per criteria laid down by Balthazar et al. Severe AP (SAP) was defined by the presence of at least one of the following criteria: (i) failure of one or more organ as defined by the Atlanta classification; (ii) an Acute Physiology and Chronic Health Evaluation II score >8; and/or (iii) CTSI >7. Patients with functional dyspepsia who attended the outpatient clinic were included as controls to estimate the normal values of anti-endotoxins and lactulose-mannitol ratio (LMR).
Management of acute pancreatitis patients: All patients were treated either in the gastroenterology ward or intensive care unit. Enteral feeding was started early and calorie intake was increased gradually from 500 to 2000 kcal/day and then continued until the patient resumed a near-normal diet. The enteral feed was prepared in the hospital kitchen and its tolerability was monitored by the treating physicians. Antibiotics were started, if (i) fever >38°C for >48 h, (ii) total leucocyte count >16,000/cumm, (iii) evidence of any extra-pancreatic infection, (iv) evidence of gas in pancreatic bed on CT scan, (v) in the second week of illness if patient continued to have features of SIRS, or (vi) when any organism was grown in culture of blood or any of the body fluids. Percutaneous drainage of collections was done in the presence of persistent SIRS or organ failure despite the use of antibiotics for at least 48 h. In patients with organ failure, organ support systems were used, as needed. Blood cultures were done at least twice a week in all patients with suspected sepsis. Drain fluid/pus was sent for cultures every alternate day. Endotracheal aspirate/sputum was sent for cultures for patients with suspected pneumonia.
Measurement of intestinal permeability: IP (LMR) was measured by urinary LM test, in the enrolled patients on day zero (D0) of their admission with a repeat assessment done on D7. Patients received 50 ml of water containing 10 g lactulose and 5 g mannitol either orally or through a nasogastric tube. Urine was collected for five hours in a receptacle with 0.2 ml of 2 per cent chlorhexidine to avoid bacterial overgrowth. The total urinary volume was measured, and two aliquots of urine (10 ml each) were immediately frozen to −70°C until analysis. Lactulose was estimated using Seliwanoff's method. Mannitol was estimated using the Corcoran and Page method. The ratio of above lactulose and mannitol values was thus computed and defined as the LMR.
Measurement of endotoxaemia:Endotoxaemia, as measured by anti-endotoxin IgG and IgM, was measured with the EndoCab ELISA kit (Hycult Biotechnology, The Netherlands). The assay is based on a solid-phase sandwich ELISA and detects and quantifies antibodies against endotoxin. The methodology used for measurement of IP and endotoxaemia was similar in all the four studies pooled for this analysis.
Statistical analysis: Data were analyzed by Statistic Software Stata 11.1 (StataCorp, Texas, USA), and presented in mean ± standard deviation/median (minimum-maximum) and frequency (%). Categorical variables were compared among groups by Chi-square/Fisher's exact test. Continuous variables following normal distribution were compared among the groups by independent t test/one-way Anova followed by post hoc comparison using Bonferroni correction. Skewed continuous variables were compared among the groups by Wilcoxon rank-sum/Kruskal-Wallis test followed by multiple comparisons using Dunn's test with Bonferroni correction.
| Results|| |
A total of 204 patients of AP who had undergone either serum anti-endotoxin antibody testing or LMR were included, of whom 180 had severe disease. Males constituted eight (33.3%) patients of mild AP, 88 (63.3%) of SAP in the first week and 25 (61%) patients of SAP presenting beyond the first week of illness (P <0.05). Baseline characteristics of all the patients are shown in [Table 1].
|Table 1: Baseline characteristics of patients with acute pancreatitis (AP)|
Click here to view
Anti-endotoxin antibodies and lactulose-mannitol ratio: [Table 2] shows anti-endotoxin IgG and IgM levels and LMR on the day of admission and D7 in patients with mild and severe AP. IgM and IgG anti-endotoxin antibodies were lower in all patients of AP compared to controls but were significantly lower in patients with mild AP and SAP within the first week of illness compared to patients with SAP presenting beyond the first week of illness. LMR was not higher in patients of SAP compared to patients of mild AP and controls.
|Table 2: Markers of endotoxaemia and intestinal permeability in patients with acute pancreatitis (AP)|
Click here to view
Anti-endotoxin antibodies and LMR with organ failure: [Table 3] shows the levels of anti-endotoxin antibodies IgG and IgM and LMR at D0 and D7 in patients of severe AP. There was no significant difference in levels of anti-endotoxin IgG and IgM among patients with organ failure compared to those without organ failure at D0 and D7. Rising trend of IgG anti-endotoxin antibodies was found in 50/74 (67.5%) patients with organ failure and in 26/55 (47.3%) patients without organ failure (P <0.05).
|Table 3: Markers of endotoxaemia and intestinal permeability in patients with severe acute pancreatitis with organ failure (n=180)|
Click here to view
In SAP patients (n=180), at baseline, the median IgG anti-endotoxin titre was 140 (4-1600) GMU/ml among patients with organ failure (n=110) and 180 (4-1760) GMU/ml among patients without organ failure (n=70), while at D7, the median IgG anti-endotoxin antibody titre was 140 (18-1600) GMU/ml in patients with organ failure compared to 174 (2-1640) GMU/ml among patients without organ failure (P < 0.05). There was no difference in LMR at D0 and D7 between patients who developed organ failure and those who did not.
Anti-endotoxin antibody titres and LMR with infection: [Table 4] shows anti-endotoxin antibody titres and LMR among patients of SAP who developed an infection. There was no significant difference in anti-endotoxin IgG and IgM and LMR among patients who developed infection subsequently compared to those who did not develop an infection.
|Table 4: Markers of endotoxaemia and intestinal permeability in patients with severe acute pancreatitis in relation to infection (n=180)|
Click here to view
Anti-endotoxin antibody titres and LMR with mortality: [Table 5] shows the association of anti-endotoxin antibody titres and LMR with mortality among patients with SAP. There was no significant difference in anti-endotoxin antibody titres and LMR in non-survivors compared to survivors.
|Table 5: Markers of endotoxaemia and intestinal permeability in patients with severe acute pancreatitis in relation to mortality (n=180)|
Click here to view
The proportion of patients with alcohol as aetiology was similar between various groups (infection vs. no infection; mortality vs. no mortality; organ failure vs. no organ failure).
| Discussion|| |
In this secondary pooled data analysis, a total of 204 patients with AP were included. Endotoxin, which is a constituent of GNB cell wall, is implicated to be involved in the development of systemic inflammatory response syndrome (SIRS) and severe sepsis. Endotoxaemia has been shown to be directly related to the severity of episodes of AP,. It is transient and may not be detected by intermittent blood sampling.
As shown in previous studies, no significant difference in anti-endotoxin IgM levels was observed between mild AP and SAP at admission. IgG anti-endotoxin antibodies, although not significantly different between mild AP and SAP at admission, were increased by a greater value and in a greater number of patients of SAP compared to mild AP over seven days. However, in patients with SAP presenting beyond the first week of illness, the median IgG and IgM anti-endotoxin antibodies were lower than controls but higher than patients of mild AP and SAP presenting within the first week of illness. Windsor et al reported no correlation between the changes in IgM anti-endotoxin antibody concentration over days and clinical outcome in patients of AP. Unlike some previous studies, but similar to Windsor et al, no significant difference in change in IgM levels was found between mild AP and SAP. Unlike Windsor et al, who checked serum IgG anti-endotoxin antibody levels for seven days from admission and reported that falling IgG titres predicted multiple organ failure, we found that rising IgG anti-endotoxin antibodies were associated more often with organ failure in patients with severe AP.
However, in a subset of patients of SAP who presented within the first week of illness, the median serum IgG anti-endotoxin antibody titre was lower at baseline and remained relatively lower than in patients without organ failure. A greater number of patients of SAP succumbed to organ failure within seven days of admission among patients with the lowest IgG anti-endotoxin levels so that their D7 anti-endotoxin titres were not available for analysis. This led to the paradoxically significant value of higher median serum IgG levels in patients without organ failure at D7. It is possible that serum IgG anti-endotoxin antibody has fallen to its nadir by the time of admission (1 to 2 wk) and that it may be rising subsequently, earlier in patients without organ failure and later and slowly in patients with organ failure who did not succumb to their organ failure. The absolute value of serum IgG anti-endotoxin antibody titre at a single point of time was not significantly different among patients with organ failure compared to patients without organ failure.
Ammori et al also reported that systemic endotoxin exposure in the early phase of AP was not associated with systemic bacterial translocation. Serum IgG and IgM anti-endotoxin titres and their changes over time were not associated with subsequent risk of infection in our study. Unlike Bose et al who reported that serum IgG and IgM anti-endotoxin titres were lower in patients with major complications, including infection, our analysis did not show an association of serum anti-endotoxin antibody titres with subsequent risk of infection. It is possible that endotoxaemia in the early phase is related to early organ failure but not associated with the risk of infection in the later phase.
Unlike some previous studies,25 which reported that serum IgG and IgM anti-endotoxin antibody titres were lower in non-survivors than in survivors, no such association was observed in the present study. IgG anti-endotoxin antibodies were lowest in mild AP and decreased further in these patients at D7. These were higher in patients with SAP and were increasing at D7 with the rising trend more often observed in patients with organ failure. There are various possible reasons for this finding. Patients with mild AP were more often admitted within one day of illness when the antibody titre might be very low. Patients with SAP were more often admitted on the fourth to fifth day of illness by which time, the trend of Ig to increase might have begun. Endotoxaemia may be only either an epiphenomenon or an aggravating factor, related to organ failure caused by a cytokine storm in the early phase, and may not be linked directly to mortality. It is also possible that the reasons for the reduction in serum IgG and IgM anti-endotoxin antibodies may not be due to binding to circulating endotoxin and subsequent removal alone but may be related to binding of antibodies to endogenous antigens, molecularly mimicking the GNB endotoxin. The subsequent increase in serum Ig against endotoxin may indicate recovery of immunity. A similar phenomenon of increasing anti-endotoxin antibodies as a marker of recovery has been proposed by Maury et al in patients with severe sepsis. This similarity is expected because of the striking similarities in cytokines and inflammatory mediator profiles in patients with SAP and sepsis.
It is reported that IP has been found to increase within the first 72 h and it correlates strongly with the clinical outcome of the patient,. Juvonen et al have suggested that after an initial increase, the IP in a patient gets restored between D8 and D45 of the illness. We found that IP, both in mild AP and SAP presenting either within or beyond the first week of illness, was similar to controls, which was in contrast to the previous findings. This could be related to later referrals to our hospital. Although our controls had higher LMR than widely reported values, they were apparently healthy people without any factor predisposing to increased gut permeability.
LMR was not significantly different among patients with organ failure, infection and mortality compared to those without these complications. A meta-analysis of 18 studies reported that gut barrier dysfunction was present in three out of five patients with AP and that the prevalence was not affected by severity. More studies are warranted to assess gut permeability, using different probes in AP.
The strength of our study was the large number of patients with severe AP and the estimation of study parameters using validated methods. However, the retrospective data analysis can lead to bias and is the major limitation of this study.
In summary, the present pooled data analysis showed that serum IgG and IgM anti-endotoxin antibodies and LMR on admission day and at day 7 were not associated with the risk of infection, organ failure and mortality in patients with AP.
Financial support & sponsorship: None.
Conflicts of Interest: None.
| References|| |
Schietroma M, Pessia B, Carlei F, Mariani P, Sista F, Amicucci G. Intestinal permeability and systemic endotoxemia in patients with acute pancreatitis. Ann Ital Chir
Koh YY, Jeon WK, Cho YK, Kim HJ, Chung WG, Chon CU, et al.
The effect of intestinal permeability and endotoxemia on the prognosis of acute pancreatitis. Gut Liver
Liang HY, Chen T, Wang T, Huang Z, Yan HT, Tang LJ. Time course of intestinal barrier function injury in a sodium taurocholate-induced severe acute pancreatitis in rat model. J Dig Dis
Zhang J, Yuan C, Hua G, Tong R, Luo X, Ying Z. Early gut barrier dysfunction in patients with severe acute pancreatitis: Attenuated by continuous blood purification treatment. Int J Artif Organs
Zou ZD, Zhang ZZ, Wang L, Wang Y, Zheng GH. The role of mesenteric lymph in pathogenesis of systemic inflammatory response syndrome and systemic complications following severe acute pancreatitis in rats. Zhongguo Wei Zhong Bing Ji Jiu Yi Xue
Huang XX, Wang XP, Ma JJ, Jing DD, Wang PW, Wu K. Effects of enteral nutrition supplemented with glutamine and arginine on gut barrier in patients with severe acute pancreatitis: A prospective randomized controlled trial. Zhonghua Yi Xue Za Zhi
Rahman SH, Ammori BJ, Holmfield J, Larvin M, McMahon MJ. Intestinal hypoperfusion contributes to gut barrier failure in severe acute pancreatitis. J Gastrointest Surg
Tarpila E, Nyström PO, Franzén L, Ihse I. Bacterial translocation during acute pancreatitis in rats. Eur J Surg
Runkel NS, Moody FG, Smith GS, Rodriguez LF, LaRocco MT, Miller TA. The role of the gut in the development of sepsis in acute pancreatitis. J Surg Res
Exley AR, Leese T, Holliday MP, Swann RA, Cohen J. Endotoxaemia and serum tumour necrosis factor as prognostic markers in severe acute pancreatitis. Gut
Kivilaakso E, Valtonen VV, Malkamäki M, Palmu A, Schröder T, Nikki P, et al.
Endotoxaemia and acute pancreatitis: Correlation between the severity of the disease and the anti-enterobacterial common antigen antibody titre. Gut
Sharma B, Srivastava S, Singh N, Sachdev V, Kapur S, Saraya A. Role of probiotics on gut permeability and endotoxemia in patients with acute pancreatitis: A double-blind randomized controlled trial. J Clin Gastroenterol
Singh N, Sharma B, Sharma M, Sachdev V, Bhardwaj P, Mani K, et al.
Evaluation of early enteral feeding through nasogastric and nasojejunal tube in severe acute pancreatitis: A noninferiority randomized controlled trial. Pancreas
Sharma M, Sachdev V, Singh N, Bhardwaj P, Pal A, Kapur S, et al.
Alterations in intestinal permeability and endotoxemia in severe acute pancreatitis. Trop Gastroenterol
Singh N, Mishra SK, Sachdev V, Sharma H, Upadhyay AD, Arora I, et al.
Effect of oral glutamine supplementation on gut permeability and endotoxemia in patients with severe acute pancreatitis: A randomized controlled trial. Pancreas
Balthazar EJ, Ranson JH, Naidich DP, Megibow AJ, Caccavale R, Cooper MM. Acute pancreatitis: Prognostic value of CT. Radiology
Bradley EL 3rd
. A clinically based classification system for acute pancreatitis. Summary of the international symposium on acute pancreatitis, Atlanta, Ga, September 11 through 13, 1992. Arch Surg
Burtis CA, Ashwon ER. Carbohydrates. In: Teitz NW, editor. Fundamentals of clinical chemistry
. Philadelphia, PA: WB Saunders Co.; 1987.p. 426-30.
Corcoran AC, Page IH. Method for the determination of mannitol in blood and urine. Fed Proc
Maury E, Blanchard HS, Chauvin P, Guglielminotti J, Alzieu M, Guidet B, et al.
Circulating endotoxin and antiendotoxin antibodies during severe sepsis and septic shock. J Crit Care
Buttenschoen K, Berger D, Hiki N, Buttenschoen DC, Vasilescu C, Chikh-Torab F. Endotoxin and antiendotoxin antibodies in patients with acute pancreatitis. Eur J Surg
Ammori BJ, Fitzgerald P, Hawkey P, McMahon MJ. The early increase in intestinal permeability and systemic endotoxin exposure in patients with severe acute pancreatitis is not associated with systemic bacterial translocation: Molecular investigation of microbial DNA in the blood. Pancreas
Windsor JA, Fearon KC, Ross JA, Barclay GR, Smyth E, Poxton I, et al.
Role of serum endotoxin and antiendotoxin core antibody levels in predicting the development of multiple organ failure in acute pancreatitis. Br J Surg
Penalva JC, Martínez J, Laveda R, Esteban A, Muñoz C, Sáez J, et al.
A study of intestinal permeability in relation to the inflammatory response and plasma endocab IgM levels in patients with acute pancreatitis. J Clin Gastroenterol
Bose SM, Verma GR, Mazumdar A, Giridhar M, Ganguly NK. Significance of serum endotoxin and antiendotoxin antibody levels in predicting the severity of acute pancreatitis. Surg Today
Wilson PG, Manji M, Neoptolemos JP. Acute pancreatitis as a model of sepsis. J Antimicrob Chemother
(Suppl A) : 51-63.
Nagpal K, Minocha VR, Agrawal V, Kapur S. Evaluation of intestinal mucosal permeability function in patients with acute pancreatitis. Am J Surg
Liu H, Li W, Wang X, Li J, Yu W. Early gut mucosal dysfunction in patients with acute pancreatitis. Pancreas
Juvonen PO, Alhava EM, Takala JA. Gut permeability in patients with acute pancreatitis. Scand J Gastroenterol
Wu LM, Sankaran SJ, Plank LD, Windsor JA, Petrov MS. Meta-analysis of gut barrier dysfunction in patients with acute pancreatitis. Br J Surg
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]