The role of ischaemia-modified albumin in the prognosis of acute pancreatitis and its correlation with the NF- κ B-mediated inflammatory response

Abstract

Objective

To investigate the correlation between the serum levels of ischaemia-modified albumin (IMA) and disease severity in rats with acute pancreatitis (AP).

Methods

A rat AP model was established and blood samples from each group were analysed at different time points. After the experiment, the pancreatic tissues of the rats were collected for pathological examination and the measurement of protein levels of NF-κB and NF-κB p65. Serum levels of amylase (α-AMY), tumour necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-8 were also compared between groups of rats.

Results

The serum IMA concentration in the severe acute pancreatitis (SAP) group was greater than that in the mild acute pancreatitis (MAP) group. The levels of the NF-κB and NF-κB p65 proteins were increased in the MAP and SAP groups in a time-dependent manner. α-AMY, TNF-α and IL-6 were increased at all time points in the MAP and SAP groups. The increases were greatest at 24 h in the SAP group. In terms of pathological changes in the pancreas, renal and lung tissues, the damage in the SAP group was more obvious than that in the MAP group.

Conclusions

Serum IMA level was associated with inflammatory markers and NF-κB p65 in rats with AP.

Keywords

Acute pancreatitis ischaemia-modified albumin IMA TNF-α NF-κB inflammatory marker

Introduction

Acute pancreatitis (AP) is one of the most clinically common acute illnesses; and is also a common acute abdominal disorder with increasing prevalence.¹ AP has the characteristics of rapid onset, rapid progression, multiple complications and high mortality. Up to 40–70% of AP patients will develop pancreatitis-related infection in the late stage; and severe cases can develop sepsis, leading to multiple organ failure.² Early identification of high-risk patients who may develop severe AP (SAP) remains a challenging task.

Existing scoring systems, such as the acute pancreatitis Bedside Severity Index (BISAP),¹ can effectively predict the severity and prognosis of AP. Other scoring systems include the Ranson score, the revised Atlanta Scale, the Acute Physiological and Chronic Health Assessment II and the Sequential Organ Failure Assessment. However, scoring systems tend to be very complex and sometimes rely on multiple surveys, which increases the time and resources required. Different biomarkers can be used to determine the severity of AP, such as C-reactive protein, serum procalcitonin and interleukin (IL)-6, which are the most commonly used biomarkers, but none of these biomarkers can accurately indicate the severity of the disease.³

In recent years, ischaemia-modified albumin (IMA) has been recognized as a valuable predictor of disease severity and has been studied and recognized as an inflammatory marker for a variety of acute and chronic diseases.^4–6 IMAs are a class of nonfunctional serum albumin with a reduced ability to bind to transition metal ions due to tissue ischaemia and are considered a candidate biomarker for acute coronary syndrome.⁷

Early activated inflammatory factors such as IL-1, tumour necrosis factor-α (TNF-α) and inflammatory mediators in AP can cause systemic inflammatory response syndrome and multiple organ failure through a series of cascaded amplification effects. The activation of the transcription factor NF-κB is a key pathway that promotes the transcription of these proinflammatory genes.⁸ Research has shown that the IMA level is correlated with AP severity, Ranson and BISAP scores and procalcitonin levels.⁹ The plasma IMA level in SAP patients is significantly greater than that in non-SAP patients, which may be related to greater permeability of the pancreatic capillary wall and a rapid decrease in blood flow after injury.¹⁰ However, the role of IMA in the prognosis of AP and its association with NF-κB-mediated inflammatory responses remains unclear.

This current study was designed to evaluate the role of IMA in the prognosis of AP and its association with NF-κB-mediated inflammatory responses and to analyse the significance of IMA as a predictive marker adjacent to classical biomarkers of inflammation or severity, such as serum amylase (α-AMY), TNF-α, IL-6 and IL-8.

Materials and methods

Materials and groups

Male Sprague-Dawley rats aged 6–8 weeks weighing 100–150 g were randomly selected for this study. The rats were subjected to a 12-h light/dark cycle, a temperature of 22 ± 1°C, a relative humidity of 60 ± 5% and standardized feeding. The rats were randomly divided into four groups. The rats were divided into a blank control group (control), a sham operation group (SO), a mild AP group (MAP) and a severe AP group (SAP).

The study was approved by the Institutional Review Board of the Animal Experiment Centre of Xinjiang Medical University (Approval number: XJBKZHR2021102001, Date: 14 October 2021). The study was conducted in accordance with the Helsinki Declaration of 1975 as revised in 2013.

Establishment of the animal model of AP

The establishment of an AP model by perfusing sodium taurocholate into the bile pancreatic duct of rats is a well-established modelling method.¹¹ The MAP and SAP groups were subjected to 2.0% and 4.0% sodium taurocholate via a 1 ml/kg retrograde cholangiopancreatinal injection, respectively. In the SO group, an equal volume of normal saline was injected into the pancreatic duct. The blank control group did not receive any treatment.

Treatment of experimental specimens

Following successful preparation of the animal model, blood samples were collected from the rats in the model groups at 0, 3, 6, 12 and 24 h after the model was generated for the detection of serum amylase and inflammatory factors. After the experiment, the morphology of the pancreas and surrounding organs was observed. The pancreas, lung and kidney tissues were cut with sterilized surgical scissors. The kidney and lung tissues were removed, fixed by soaking in 4% paraformaldehyde and sent to the pathology room for preparation of sections. The tail tissue of the pancreas was soaked in dilute formaldehyde (intended to be a paraffin specimen) and other pancreatic tissue was frozen in liquid nitrogen at –70°C for subsequent Western blot analysis.

ELISA determination of α-AMY, IMA and inflammatory cytokines

The serum levels of α-AMY, TNF-α, IL-6 and IL-8 were determined by enzyme-linked immunosorbent assays (ELISAs). Rat serum samples were collected in serum separation tubes and stored at room temperature for 2 h or overnight at 4°C. The samples were centrifuged at 1000 g for 20 min at 4°C and the supernatant was collected (H2050R high-speed refrigerated centrifuge; Xiangyi Power Testing Instruments, Changsha, China). The supernatant was either used immediately or stored at –80°C. To measure the serum levels of α-AMY, the ELISA microplate reader was preheated (Alpha AMY assay kit; Kaiao Technology Development, Beijing, China) for 30 min. All reagents were thawed to room temperature; and then the serum samples, distilled water, standards and ELISA kit reagents were added in sequence to a 96 well plate, mixed well and the absorbance value was read at 405 nm at 37°C for 1 min. To measure the serum levels of TNF - α, IL-6, and IL-8 determination, 100 μl of the serum sample solution or different concentration standards were added to each well of the microplate; and 100 μl of universal diluent was added to the blank well. The microplate was covered with a sealing membrane and incubated at 37°C for 2 h. The wells were washed five times at 4°C to wash away any unbound substances using 300 ml of concentrated wash buffer (0.1 mmol phosphate-buffered saline; pH 7.2–7.4) from the TNF-α, IL-6 and IL-8 ELISA kits diluted with deionized water (Jianglai Biotechnology, Shanghai, China). An enzyme-linked polyclonal antibody specific for TNF-α, IL-6 and IL-8 were added to the wells and incubated for 2 h at room temperature. Following a wash using the same wash buffer to remove any unbound antibody–enzyme complex, a substrate solution was added to the wells and incubated for 2 h at room temperature. The intensity of the colour was measured at 450 nm on a microplate reader (K6600A ELISA reader; Kaiao Technology Development).

The measurement of IMA uses a double antibody one-step sandwich ELISA method. The serum sample, standards and horseradish-peroxidase (HRP)-labelled detection antibody were added to the precoated IMA antibody micropores in sequence, incubated and thoroughly washed (YJ574832b, IMA ELISA detection kit; Shanghai Enzyme Linked Biotechnology, Shanghai, China). The substrate 3,3′,5,5′ tetramethylbenzidine is converted to blue under the catalysis of peroxidase and to the final yellow under the action of acid. The absorbance was measured at a wavelength of 450 nm using a microplate reader (K6600A ELISA reader; Kaiao Technology Development) and the sample concentration was calculated using the standard solutions.

The detectable concentration range for each antigen was as follows: α-AMY (50–1200 U/l); IMA (1.78–120 U/ml); TNF-α (3.12–200 pg/ml), IL-6 (3.12–200 pg/ml); and IL-8 (7.5–240 pg/ml). Intra- and interassay coefficients of variation for all ELISAs were <8% and <12%, respectively.

Histopathological examinations

Rat pancreatic tissue was fixed with 10% neutral buffered formalin and tissue sections stained with haematoxylin and eosin (H&E) were prepared. According to the Schmidt method, pathological grading of the lungs and kidneys was performed and evaluated by two senior pathologists (B.Z. & T.A.) in a double-blinded manner as follows: (i) take the upper lobe of the right lung for routine pathological examination and refer to the Osman lung histological scoring standard for lung injury scoring; (ii) take a portion of kidney tissue and fix in paraformaldehyde followed by routine embedding, sectioning and H&E staining so that any pathological changes can be observed under a upright microscope (Leica DM4000; Leica Microsystems Trading, Shanghai, China) with a field of view of ×200 times.

The NF-κB and NF-κB p65 protein levels in pancreatic tissue were detected by Western blotting

Western blotting analysis was performed using standard techniques as follows. An appropriate amount of pancreatic tissue was cut and placed in a centrifuge tube. Lysis buffer (1 ml RIPA plus 10 ml PMSF; Beyotime Biotech, Shanghai, China) was added and the tissue was crushed using ultrasonication in an ice bath. Then the sample was centrifuged (H2050R high-speed refrigerated centrifuge; Xiangyi Power Testing Instruments) at 12000 g for 5 min at 4°C and the supernatant was retained. Protein concentration was determined using a BCA protein assay reagent kit (P0012; Beyotime). The proteins (50– 200 mg per lane) were separated by 12% sodium dodecyl sulphate–polyacrylamide gel electrophoresis. 90 min. The proteins were then transferred to polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). The membranes were incubated in 3% bovine serum albumin blocking solution (neoFroxx GmbH, Einhausen, Germany) for 1 h at 37°C. The membranes were then incubated with rabbit antirat antibodies against NF-κB, NF-κB p65 and β-actin (1:10000 dilution; Bioss, Beijing, China) overnight at 4°C; then washed three times with Tris-buffered saline-Tween 20 (TBST; pH 7.5; 20 mmol/l Tris–HCl, 150 mmol/l sodium chloride, 0.1% Tween-20) for 5 min at room temperature. The membranes were then incubated with the appropriate goat antirabbit IgG-HRP-conjugated secondary antibodies (1:20000 dilution; EarthOx, Burlingame, CA, USA) for 1 h at 37°C followed by washing three times with TBST (pH 7.5) for 5 min at room temperature. The membranes were developed using an enhanced chemiluminescence reagent substrate (GE Healthcare Biosciences, Piscataway, NJ, USA) and exposed strip from a Kodak Image Station 4000 MM (Kodak, Rochester, New York, USA).

Statistical analyses

All statistical analyses were performed using GraphPad Prism 9.5 (Graphpad Software Inc., San Diego, CA, USA). Categorical data are expressed as a percentage (%) and compared using χ²-test. Continuous data were tested for normality and are expressed as the mean ± SD. For continuous data with homogeneity of variance, the independent sample t-test was used for comparisons between two groups. Pearson correlation analysis was used to determine whether the α-AMY concentration was correlated with the surface level of the NF-κB p65 protein. All of the data presented in this article were subjected to at least three independent experiments. A P-value <0.05 was considered statistically significant.

Results

This current study investigated 60 rats that were evenly distributed among four groups. There were nine deaths in the MAP and SAP groups and there were an additional 11 failed models, so 10 rats were available for experimentation in each group. Pathological analysis revealed that the gross morphology of the pancreas in the MAP and SAP groups showed obvious oedema, haemorrhage, necrosis and extensive necrosis of the peripancreatic adipose tissue, which indicated successful modelling of AP (Figure 1). Prolongation of time from 6 h to 24 h increased the pancreatic injury. At each time point, the degree of pancreatic injury and necrosis of the SAP group was more obvious than the MAP group. There was no significant difference in the serum IMA levels at 24 h between the sham operation group and the control group (Figure 2(a)). The levels of serum IMA in the MAP and SAP groups increased at each time point (6–24 h). The IMA levels in the MAP and SAP groups were significantly increased at each time point compared with the control group (P < 0.05 for all comparisons).

Figure 1.

Representative images of the gross morphology of the pancreas in the different groups of rats (control and sham at 24 h): (a) control group; (b) MAP 6 h group; (c) MAP 12 h group; (d) MAP 24 h group; (e) SO group; (f) SAP 6 h group; (g) SAP 12 h group and (h) SAP 24 h group. MAP, mild acute pancreatitis; SO, sham operation; SAP, severe acute pancreatitis. The colour version of this figure is available at: http://imr.sagepub.com.

Figure 2.

(a) Serum levels of ischaemia-modified albumin (IMA) in the different groups of rats (control and sham at 24 h). **P < 0.01; ***P < 0.001; independent sample t-test and (b) Pearson correlation analysis results for the correlation between amylase (α-AMY) and NF-κB p65 levels (R = 0.760; P < 0.001). The colour version of this figure is available at: http://imr.sagepub.com.

Pearson correlation analysis indicated a positive correlation between α-AMY and NF-κB p65 expression (R = 0.664; P < 0.001) (Figure 2(b)). The serum levels of α-AMY, TNF-α, IL-6 and IL-8 in the different groups of rats were detected using ELISA kits. Compared with the levels in the control group, the α-AMY, TNF-α and IL-6 levels in the MAP and SAP groups were increased at all time points; and some of the increases were significant (P < 0.05 for comparisons) (Figure 3). Compared with the levels in the control group, the IL-8 levels in the MAP and SAP groups were increased at some time points; and some of the increases were significant (P < 0.05 for comparisons).

Figure 3.

Serum levels of amylase (α-AMY), tumour necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-8 in the different groups of rats (control and sham at 24 h). Each sample was duplicated and the figure is representative of three independent assays (n = 5). *P < 0.05; **P < 0.01; ***P < 0.001; independent sample t-test. The colour version of this figure is available at: http://imr.sagepub.com.

The levels of NF-κB and NF-κB p65 protein in pancreatic tissue samples from all groups of rats was measured using Western blot analysis (Figure 4). Compared with the levels in the control group, the levels of NF-κB and NF-κB p65 protein in the MAP group at 24 h were significantly increased; and those in the SAP group at 12 h and 24 h were significantly increased (P < 0.05 for comparisons).

Figure 4.

Western Blot analysis of the levels of NF-κB and NF-κB p65 protein in pancreatic samples of rats in each group at 6 h, 12 h and 24 h (control and sham at 24 h). *P < 0.05; **P < 0.01; ***P < 0.001; independent sample t-test. CO, control; SO, sham operation; MAP, mild acute pancreatitis; SAP, severe acute pancreatitis; kDa, kilodalton. The colour version of this figure is available at: http://imr.sagepub.com.

Histopathological examination of the pancreas specimens showed that islet cells in the control and SO groups demonstrated the following characteristics: normal distribution, round nucleus, cytoplasmic distribution, clear structure and no degeneration and necrosis (Figure 5). Compared with the control group, pancreatic cell oedema, inflammatory cell infiltration, bleeding and necrosis were not obvious in the MAP 6 h group and SAP 6 h groups. The MAP 12 h and SAP 12 h groups showed obvious inflammatory manifestations as follows: widening of the interlobular space, interlobular oedema, obvious inflammatory cell infiltration, acinous cell swelling and a small amount of haemorrhage and necrosis. The MAP 24 h and SAP 24 h groups showed extensive coagulation necrosis with bleeding, unclear cell structure in the necrotic areas and a large number of inflammatory cells infiltrating the parenchyma; with the pancreases of the SAP 24 h group being the most severely damaged.

Figure 5.

Histopathological examination of the pancreas in the different groups of rats (control and sham at 24 h): (a) control group; (b) MAP 6 h group; (c) MAP 12 h group; (d) MAP 24 h group; (e) SO group; (f) SAP 6 h group; (g) SAP 12 h group and (h) SAP 24 h group. MAP, mild acute pancreatitis; SO, sham operation; SAP, severe acute pancreatitis. Scale bar 100 µm; haematoxylin and eosin. The colour version of this figure is available at: http://imr.sagepub.com.

Histopathological examination of the renal tissue structure was undertaken in the different groups of rats including the control and SO groups (Figure 6). In the MAP 6 h and SAP 6 h groups, the glomerular structure was still intact, the renal tubules were slightly swollen and the interstitium was congested. In the MAP 12 h and SAP 12 h groups, the renal glomeruli showed swelling, structural damage, gradual disordered arrangements, significant swelling of the renal tubules, narrowing of the renal lumen, congestion of the renal interstitium and infiltration by inflammatory cells. The MAP 24 h and SAP 24 h groups showed renal glomerular swelling, capillary network cracks, widespread renal tubular lesions, disordered arrangements, swelling, almost no narrowing of the lumen, epithelial cell necrosis, structural disappearance, severe renal interstitial congestion and significantly increased inflammatory cell infiltration.

Figure 6.

Histopathological examination of the renal tissue in the different groups of rats (control and sham at 24 h): (a) control group; (b) MAP 6 h group; (c) MAP 12 h group; (d) MAP 24 h group; (e) SO group; (f) SAP 6 h group and (g) SAP 12 h group; (h) SAP 24 h group. MAP, mild acute pancreatitis; SO, sham operation; SAP, severe acute pancreatitis. Scale bar 100 µm; haematoxylin and eosin. The colour version of this figure is available at: http://imr.sagepub.com.

Histopathological examination of the lung tissue structure was undertaken in the different groups of rats including the control and SO groups (Figure 7). Compared with the control group, pathological changes such as inflammatory cell infiltration, alveolar wall oedema and bleeding were observed in the lung tissue of rats in each model group. At the same time point, the pathological changes in the SAP groups were more significant than those in the MAP groups. The extent of the pulmonary oedema, inflammatory cell infiltration and bleeding in the MAP 12 h and SAP 12 h groups was worse than that observed in the MAP 6 h and SAP 6 h groups. Similarly, the extent of the oedema, infiltration of inflammatory cells and bleeding of the lung tissue in the MAP 24 h and SAP 24 h groups was more severe than that observed in the MAP 12 h and SAP 12 h groups. The Osman scores for lung injury in the different groups of rats demonstrated that the scores were highest in the SAP 12 h and SAP 24 h groups, followed by the MAP 24 h group; and there were significant differences compared with the control and SO groups (P < 0.001) (Table 1).

Figure 7.

Histopathological examination of the lung tissue in the different groups of rats (control and sham at 24 h): (a) control group; (b) MAP 6 h group; (c) MAP 12 h group; (d) MAP 24 h group; (e) SO group; (f) SAP 6 h group; (g) SAP 12 h group and (h) SAP 24 h group. MAP, mild acute pancreatitis; SO, sham operation; SAP, severe acute pancreatitis. Scale bar 100 µm; haematoxylin and eosin. The colour version of this figure is available at: http://imr.sagepub.com.

Table 1.

The Osman lung injury scores for the different groups of rats investigated in a study designed to evaluate the role of ischaemia-modified albumin in the prognosis of acute pancreatitis and its association with NF-κB-mediated inflammatory responses.

Group	Osman score
Control (n = 5)	0.00 ± 0.00
SO (n = 5)	0.00 ± 0.00
MAP 6 h (n = 5)	2.33 ± 0.58
MAP 12 h (n = 5)	2.33 ± 0.58
MAP 24 h (n = 5)	3.66 ± 0.58
SAP 6 h (n = 5)	3.33 ± 0.58
SAP 12 h (n = 5)	4.00 ± 0.00
SAP 24 h (n = 5)	4.00 ± 0.00
F	50.07
P-value	P < 0.001

Data presented as mean ± SD.

SO, sham operation; MAP, mild acute pancreatitis; SAP, severe acute pancreatitis.

Discussion

The current research findings demonstrated that IMA was correlated with AP disease severity. In addition, some laboratory parameters, including α-AMY, TNF-α and IL-6, were correlated with AP disease severity. The levels of α-AMY were positively correlated with NF-κB p65.

Ischaemic processes are thought to contribute to the development of AP and ischaemic markers help diagnose and determine the prognosis of this disease.¹² IMA is produced by the liver and is associated with interactions between free radicals produced during ischaemia.¹³ Research suggests that the formation of IMA is the earliest sign of ischaemia and is related to vascular endothelial disorders in the early stage of the disease.¹⁴ Microthrombosis is associated with proinflammatory cytokines, prothrombotic autoantibodies and an impaired endothelial barrier; and may lead to local tissue ischaemia, oxidative stress and increased albumin oxidation in SAP patients.¹⁵ IMA levels are also elevated in patients with COVID-19, deep vein thrombosis and pulmonary embolism.^16,17

A previous study reported that IMA increases the incidence of AP,¹⁸ but its effectiveness in terms of prognosis is limited to exploratory studies. The relationship between the two has not been fully evaluated in clinical trials. Some studies have also reported that the common background of acute and chronic inflammatory states, oxidative stress and reactive oxygen species formation leads to increased serum IMA concentrations in patients with noncardiac diseases (such as sepsis, intussusception ischaemia and rheumatoid arthritis).^19,20 This current study investigated the value of IMA as a prognostic marker in patients with AP, determined whether IMA is effective in assessing disease severity and assessed the association between IMA and other routine clinical inflammatory and prognostic markers.

Acute pancreatitis is often accompanied by cytokine changes, especially in SAP, where the activation of NF-κB in the pancreas itself, distant organs, tissues and intracellular cells is significantly enhanced, accompanied by the upregulation of various inflammatory cytokines, which may play an important role in the occurrence and development of SAP.^21–23 NF-κB exists in the body as a dimer composed of Rel, p65, p50 and other molecules and it can regulate the expression of inflammatory mediators such as TNF-α, IL-1, IL-6 and IL-8; and in doing so it can regulate the cytokine-mediated inflammatory response.²⁴ The dominant dimer in the pancreas is the NF-κB p65/p50 dimer.²⁵ In the classical inflammatory pathway, the NF-κB dimer travels to the nucleus and binds to a common sequence of transcription factors and target genes. Experimentally induced AP has been reported to lead to the apoptosis of pancreatic acinar cells and increased NF-κB activation.²⁶ A previous study suggested that NF-κB activation was related to the severity of AP.²⁷ Another study reported that exposure to cyanin led to apoptosis and increased the expression of NF-κB p65 in pancreatic cells.²⁸ Similarly, the expression level of NF-κB p65 and apoptosis of pancreatic cells were increased in the pancreatic tissues of rats with SAP.²⁹

The current study demonstrated that IMA increased at all time points in the MAP and SAP groups of rats, which might be related to the high permeability of pancreatic capillary endothelial vessels and the rapid decrease in the effective circulating blood volume after damage, indicating that AP can lead to ischaemic changes. The IMA levels in the two AP model groups increased with time, which was considered to be related to the prolonged duration of tissue hypoxia, hypoperfusion, inflammation and oxidative damage. The IMA level was significantly elevated in the SAP group at all time points compared with the control group, which was associated with more severe pancreatic injury. The current findings suggest that α-AMY, TNF-α and IL-6 play a role in determining the severity of pancreatitis. The α-AMY levels tended to increase with increasing AP severity and time, while the corresponding NF-κB p65 protein level also gradually increased, and the two biomarkers were positively correlated. The histopathological findings in the current study demonstrated that there were varying degrees of pathological changes in the pancreatic, renal and lung tissues of rats in the MAP and SAP groups compared with the control group. The main manifestation was that with an increase in the modelling time, the extent of inflammatory cell infiltration increased and cell necrosis was accompanied by bleeding. The pathological changes in the SAP groups were more significant than those in the MAP groups. The Osman scores for lung injury in the different groups of rats demonstrated that the scores were highest in the SAP 12 h and SAP 24 h groups, followed by the MAP 24 h group; and there were statistically significant differences compared with the control and SO groups. These current findings suggest that the release of inflammatory factors in SAP (e.g. TNF-α, IL-6 and IL-8) result in significant damage to multiple organs in the body. Even in rats in the MAP groups had a degree of damage to multiple organs and the damage worsened over time. Since pancreatic tissue sampling was limited to pancreatic tail tissue, the degree of NF-κB activation in cells in other parts of the pancreas cannot be explained. However, further studies on the influence of other organs on ischaemia-sensitive target organs are needed.

This current study had a several limitations. First, the study only used rats as experimental subjects and there may be important differences between animal models and human diseases. The effectiveness and accuracy of the results in human clinical applications need further verification. Secondly, some inflammatory markers have shortcomings in determining the severity of AP. For example, the IMA levels continued to increase up to the 24-h time point, but as that was the final time point in this current study, it was not possible to determine when the highest IMA value was reached in relation to the status of AP in this animal model. Thirdly, studies have confirmed the correlation between ischaemic markers, IMA and inflammatory markers, as well as with NF-κB p65 expression. However, there is still insufficient research on the correlation between endothelial dysfunction and IMA caused by ischaemia in AP. Further in-depth research is required.

In conclusion, these current results suggest that the serum IMA level was associated with inflammatory markers and NF-κB p65 in rats with AP. However, further studies are needed to explore the mechanisms underlying this association, particularly the association between IMA and impaired vascular endothelial disorders in AP and the potential role of IMA as a candidate biomarker for AP.

Footnotes

Acknowledgements

We acknowledge the support of the Natural Science Foundation of Xinjiang Uygur and the hard work of all co-authors on this research project.

Author contributions

Xinjian Xu conceived and designed the study. Junxiang Zhang and Shixing Wu performed the laboratory experiments, then drafted the manuscript. Cheng Geng guided the experiments and revised the manuscript. Bolin Zhang, Xinxin Tian, Tigu A and Hongde Su analysed the data and interpreted the results. Tigu A and Hongde Su created the figures and table. All authors approved the final version of the manuscript.

Availability of data and materials

The datasets generated and/or analysed during the current study are available from the corresponding author on reasonable request.

Declaration of conflicting interest

The authors declare that there are no conflicts of interest.

Funding

This work was supported by the Natural Science Foundation of Xinjiang Uygur Autonomous Region (), China (no. 2022D01C311). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

ORCID iD

Xinjian Xu

References

Mederos

Reber

Girgis

MD.

Acute Pancreatitis: A Review. JAMA 2021; 325: 382–390.

Wang

Zhang

Liu

, et al. Early predictive value of presepsin for secondary sepsis and mortality in intensive care unit patients with severe acute pancreatitis. Shock 2023; 59: 560–568.

Behera

Mishra

Sahu

, et al. C-reactive protein/albumin and ferritin as predictive markers for severity and mortality in patients with acute pancreatitis. Prz Gastroenterol 2023; 18: 168–174.

Hakkoymaz

Nazik

Seyithanoğlu

, et al. The value of ischemia-modified albumin and oxidative stress markers in the diagnosis of acute appendicitis in adults. Am J Emerg Med 2019; 37: 2097–2101.

Ellidag

Eren

Yilmaz

, et al. Oxidative stress and ischemia-modified albumin in chronic ischemic heart failure. Redox Rep 2014; 19: 118–123.

Ralapanawa

Sivakanesan

Tennakoon

, et al

. Ischemia-modified albumin: is it a promising marker in acute coronary syndrome?

BMC Cardiovasc Disord 2024; 24: 436.

Mangoni

Zinellu

Serum Concentrations of Ischaemia-Modified Albumin in Acute Coronary Syndrome: A Systematic Review and Meta-Analysis. J Clin Med 2022; 11: 4205.

Zhang

Zerumbone ameliorates the inflammatory response and organ damage in severe acute pancreatitis via the ROS/NF-κB pathway. BMC Gastroenterol 2023; 23: 333.

Yarkici

Derviş Hakim

Arı

, et al. Can Ischemia Modified Albumin (IMA) Levels Be a Predictor of Acute Pancreatitis? Turk J Gastroenterol 2021; 32: 327–335.

10.

Zhang

Fan

, et al. Prognostic significance of ischemia-modified albumin for severe acute pancreatitis. Int J Clin Exp Pathol 2017; 10: 10552–10558.

11.

Ding

Chen

, et al . Soluble Fibrinogen-Like Protein 2 Downregulation and Th17/Treg Imbalance in a Taurocholate-Induced Murine Experimental Model of Severe Acute Pancreatitis. J Clin Lab Anal 2024; 38: e25076.

12.

Winterbourn

Bonham

Buss

, et al. Elevated protein carbonyls as plasma markers of oxidative stress in acute pancreatitis. Pancreatology 2003; 3: 375–382.

13.

Gidenne

Ceppa

Fontan

, et al. Analytical performance of the Albumin Cobalt Binding (ACB) test on the Cobas MIRA Plus analyzer. Clin Chem Lab Med 2004; 42: 455–361.

14.

Balta

Endothelial Dysfunction and Inflammatory Markers of Vascular Disease. Curr Vasc Pharmacol 2021; 19: 243–249.

15.

Kurt

Demirci

Alan

Endothelial Dysfunction and Ischemia-Modified Albumin Levels in Males with Diabetic and Nondiabetic Erectile Dysfunction. Dis Markers 2022; 2022: 3661822.

16.

Erre

Bosincu

Faedda

, et al. Antiphospholipid syndrome nephropathy (APSN) in patients with lupus nephritis: a retrospective clinical and renal pathology study. Rheumatol Int 2014; 34: 535–541.

17.

Saglam

Sener

Bayrak

, et al. Analysis of Ischemia-Modified Albumin (IMA) and Coagulation Parameters in Patients with SARS-CoV-2 Pneumonia. J Clin Med 2023; 12: 4304.

18.

Chawla

Loomba

Guru

, et al. Ischemia Modified Albumin (IMA) – A Marker of Glycaemic Control and Vascular Complications in Type 2 Diabetes Mellitus. J Clin Diagn Res 2016; 10: BC13–BC16.

19.

Erre

Chessa

Bassu

, et al. Association between ischemia-modified albumin (IMA) and peripheral endothelial dysfunction in rheumatoid arthritis patients. Sci Rep 2024; 14: 3964.

20.

Shevtsova

Gordiienko

Tkachenko

, et al. Ischemia-Modified Albumin: Origins and Clinical Implications. Dis Markers 2021; 2021: 9945424.

21.

Jin

Yang

Zhao

, et al. Apocynin alleviates lung injury by suppressing NLRP3 inflammasome activation and NF-κB signaling in acute pancreatitis. Int Immunopharmacol 2019; 75: 105821.

22.

Uluışık

Keskin

Özaydın

, et al. Ameliorative effects of the melatonin on some cytokine levels, NF-κB immunoreactivity, and apoptosis in rats with cerulein-induced acute pancreatitis. Iran J Basic Med Sci 2024; 27: 279–285.

23.

Duan

, et al. Author Correction: Inhibition of RIPK1-dependent regulated acinar cell necrosis provides protection against acute pancreatitis via the RIPK1/NF-κB/AQP8 pathway. Exp Mol Med 2024; 56: 491–493.

24.

Feng

Tong

, et al. Saikosaponin a attenuates hyperlipidemic pancreatitis in rats via the PPAR-γ/NF-κB signaling pathway. Exp Ther Med 2020; 19: 1203–1212.

25.

Pan

Wang

, et al. Augmenter of liver regeneration (ALR) regulates acute pancreatitis via inhibiting HMGB1/TLR4/NF-κB signaling pathway. Am J Transl Res 2018; 10: 402–410.

26.

Chen

Sun

, et al. Hydrogen-rich saline ameliorates the severity of l-arginine-induced acute pancreatitis in rats. Biochem Biophys Res Commun 2010; 393: 308–313.

27.

Huang

Liu

Daniluk

, et al. Activation of nuclear factor-κB in acinar cells increases the severity of pancreatitis in mice. Gastroenterology 2013; 144: 202–210.

28.

Bansod

Godugu

Nimbolide ameliorates pancreatic inflammation and apoptosis by modulating NF-κB/SIRT1 and apoptosis signaling in acute pancreatitis model. Int Immunopharmacol 2021; 90: 107246.

29.

Zhang

, et al. Colchicine improves severe acute pancreatitis-induced acute lung injury by suppressing inflammation, apoptosis and oxidative stress in rats. Biomed Pharmacother 2022; 153: 113461.