|Year : 2022 | Volume
| Issue : 2 | Page : 170-174
Impact of Proinflammatory Cytokines: (Interleukin 6, Interleukin 1α, and Interleukin 1β) on Biochemical Parameters in Severe Acute Respiratory Syndrome Coronavirus 2 Patients in Iraq
Ismael Raheem Al-Muhana1, Mohanad Mohsin Ahmed2, Ishaq Raheem Al-Muhana3, Baraa Akeel AL-Hasan4
1 Department of Microbiology, College of Veterinary Medicine, Kufa University, Najaf, Iraq
2 Department of Microbiology, College of Medicine, Karbala University, Karbala, Iraq
3 Department of Veterinary Genetic, Institute of Health sciences, Ankara University, Ankara, Turkey
4 Department of Medical Laboratory Technology, College of Medical Technology, The Islamic University, Najaf, Iraq
|Date of Submission||30-Sep-2021|
|Date of Acceptance||16-Nov-2021|
|Date of Web Publication||17-Jun-2022|
Ismael Raheem Al-Muhana
Department of Microbiology, College of Veterinary Medicine, Kufa University, Najaf
Source of Support: None, Conflict of Interest: None
Background: The cytokine storm and specific biochemical results were both observed during Coronavirus disease 2019 (COVID-19) infection. The aim of this study was to see if there was any correlation between biochemical findings and cytokine levels. Methods: A total of 65 COVID-19 patients, including 32 males and 33 females with ages between (16 and 90 years) were enrolled in this study. Between December 2020 and February 2021, these patients were admitted to Al-Amal Specialized Hospital or ward of COVID-19 care in Al-Sader Hospital. These patients were classified into severe/critical (43 cases), mild/moderate (22 cases) according to the guidelines released by National Health World depending on SpO2 percentage. The biochemical indices were measured using FUJI DRI-CHEM NX500 automated clinical chemistry analyzer (FUJIFILM, Japan). The proinflammatory cytokine (interleukin-6 [IL-6], IL-1α, and IL-1β) were measured using ELISA technique. Results: IL-6 serum levels were negatively correlated with SpO2 (P = 0.002, R = - 0.372) and serum albumin (P = 0.034, R = - 0.301). IL-1α serum levels showed significant negative correlation with serum albumin levels (P = 0.039, R = - 0.259). Furthermore, a positive correlation found between IL-1β serum with serum levels of both AST and LDH (P = 0.049; R = 0.255; P = 0.054; R = 0.320 respectively). Discussion: Increased IL-6 serum levels have a direct impact on SpO2 percentage and serum albumin. Whereas the elevated levels of both IL-1α and IL-1β are a possible cause of acute inflammation and liver damage in COVID-19. Conclusions: This study further confirms the growing evidence on the direct role of proinflammatory cytokines in the biochemical changes shown in COVID-19.
Keywords: Biochemical parameters, COVID-19, proinflammatory cytokines
|How to cite this article:|
Al-Muhana IR, Ahmed MM, Al-Muhana IR, AL-Hasan BA. Impact of Proinflammatory Cytokines: (Interleukin 6, Interleukin 1α, and Interleukin 1β) on Biochemical Parameters in Severe Acute Respiratory Syndrome Coronavirus 2 Patients in Iraq. Biomed Biotechnol Res J 2022;6:170-4
|How to cite this URL:|
Al-Muhana IR, Ahmed MM, Al-Muhana IR, AL-Hasan BA. Impact of Proinflammatory Cytokines: (Interleukin 6, Interleukin 1α, and Interleukin 1β) on Biochemical Parameters in Severe Acute Respiratory Syndrome Coronavirus 2 Patients in Iraq. Biomed Biotechnol Res J [serial online] 2022 [cited 2022 Jun 26];6:170-4. Available from: https://www.bmbtrj.org/text.asp?2022/6/2/170/347709
| Introduction|| |
Coronavirus disease 2019 (COVID-19) is a highly contagious respiratory infection that was first discovered in Wuhan, China, in December 2019. It caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a virus with single positive-strand RNA, having glycoproteins projection on envelopes so reveals a halo-like appearance or corona. The disease has spread over the world, and the World Health Organization declared it a pandemic on March 11, 2020. The source of COVID-19 is unknown; however, the first documented case was related to the Huanan seafood market in Wuhan City. It is thought to have been spread zoonotically, in a Wuhan wet market where meat and game animals were sold. Despite the fact that the majority of cases were mild to moderate, the rising number of COVID-19 instances resulted in a considerable proportion of patients experiencing severe symptoms and dying. Patients infected with SARS-CoV-2 are likely to experience several stages of the disease, ranging from asymptomatic-to-mild symptoms such as dry cough, sore throat, and fever. The majority of cases have spontaneously resolved. Organ failure, septic shock, pulmonary edema, severe pneumonia, and acute respiratory distress syndrome (ARDS) have all been reported as fatal consequences in some patients. Many laboratory parameters, such as hypoalbuminemia, elevated liver enzymes, creatinine, and nonspecific inflammatory markers such as C-reactive protein (CRP), lactate dehydrogenase (LDH), elevated D-dimer, and ferritin can be used to assess the severity of the disease and predict its unfavorable course. The most critically ill patients may experience a cytokine storm, which is characterized by a surge in the production of many cytokines that cause long-term damage and fibrosis of lung tissue. Evidence suggest that proinflammatory cytokines play a key role in the pathogenesis of COVID-19 patients. When the anti-inflammatory reaction is out of control, it can lead to lung tissue involvement and ARDS, as well as a systemic response that damages several organs. The majority of previous studies have solely looked at the role of interleukin-6 (IL-6) in COVID-19. Other cytokines in COVID-19, particularly the proinflammatory cytokine, have received little attention in terms of their properties and roles. The relationship between proinflammatory cytokine (IL-6, IL-1α, and IL-1β) and biochemical results in COVID-19 patients were investigated in this study. The goal of this study was to see how proinflammatory cytokines affected biochemical markers in COVID-19 patients of various levels of severity.
| Methods|| |
This study enrolled a total of 65 COVID-19 patients, 32 males and 33 females. All patients were admitted to Al-Amal Specialized Hospital or Al-Sader Hospital's COVID-19 ward. Their ages ranged from 16 to 19 years. The diagnosis COVID-19 in each patient in this study was confirmed by SARS-CoV-2-positive reverse transcriptase–polymerase chain reaction result. Patients were grouped based on the SpO2 percentage into the mild/moderate group (<90%–94%) and severe/critical group (<90%) according to the guidelines released by National Health World.
The Najaf Health Directorate granted ethical approval. Before taking the sample, the patients and his relative were asked for their permission. When sampling, health and safety precautions were implemented.
Sample collection and processing
Blood samples and serum were collected from each patients which used to determine both the proinflammatory cytokine serum levels and biochemical tests. The proinflammatory cytokine (IL-6, IL-1α, and IL-1β) measured using ELK Company ELISA kits and a Biotek EL800 automated immunoassay analyzer (BioTek, USA). While the biochemical indices were measured using FUJI DRI-CHEM NX500 automated clinical chemistry analyzer (FUJIFILM, Japan). CRP was detected by immunoturbidimetry method. Furthermore, D-dimer and ferritin were measured using MINI VIDAS Compact multiparametric immunoanalyzer (bioMérieux, France).
To perform statistical analysis, Data were entered into version 26 of SPSS for Windows (GraphPad Software, San Diego, California, USA). The results were expressed as mean ± standard deviation (SD). A P < 0.05 was considered to indicate the statistical significance and highly significant if P < 0.001. In addition, the relationship between proinflammatory cytokine serum levels and biochemical tests was explained using Pearson correlation.
| Results|| |
Demographic and clinical characteristic of Coronavirus disease 2019 patients
[Table 1] summarizes the demographic and clinical features of COVID-19 patients. The patients were classified into mild/moderate group and severe/critical group according to SpO2 percentage. Among them, 32 (49.2%) were males and 33 (50.7%) were females. The mean age of total patients was 55.71 years (range, 16–90 years). The severe/critical group of COVID-19 patients in comparison to mild/moderate group are older age (mean ± S. D. 62.58 ± 17.409) and more likely to have high systolic blood pressure and high blood glucose level. Cough (18.8%) and fever (18.8%) were the most common symptoms in the mild/moderate group (22.7%). Meanwhile, Cough 41 (95.3%) and respiratory distress 37 (86.04%) were the most common symptoms in the severe/critical group, respectively.
|Table 1: The demographic and clinical characteristic of coronavirus disease-2019 patients|
Click here to view
The correlation between immunological and biochemical parameters
[Table 2] shows negative correlation between SpO2 percentage and IL-6 level (P = 0.002; R = −0.372) as shown in [Figure 1]. This implies that IL-6 may have a direct impact on SpO2 percentage in severe COVID-19 patients. In same way, there was a positive correlation between IL-6 level and creatinine level (P = 0.012; R = 0.309). This indicates that IL-6 may have a direct effect on creatinine level. On the other hand, there was negative correlation between the level of albumin and increased levels of both IL-6 and IL-1α (P = 0.034; R = −0.301; P = 0.039; R = −0.259, respectively). This means that both cytokines may have a direct impact on the albumin level.
|Table 2: Correlation between cytokines and biochemical parameter in patients with coronavirus disease-2019|
Click here to view
|Figure 1: Scatterplot of SpO2 percentage in relation to serum interleukin-6 pg/ml. Pearson correlation coefficient: r = −0.372** (P = 0.002)|
Click here to view
The increase in the level of alkaline phosphatase (ALP) was positively correlated with increased level of IL-1α (P = 0.024; R = 0.304). This indicates that of IL-1α may have a direct effect on the level of ALP. In the same way, the increased level of aspartate aminotransferase (AST) was positively correlated with increased level of IL-1β (P = 0.049; R = 0.255). This indicates that IL-1β may have a direct impact on AST level. In [Figure 2], there was positive correlation between the level of IL-1β and LDH level (P = 0.054; R = 0.320). This indicates that IL-1β may have a direct effect on the LDH.
|Figure 2: Scatterplot lactate dehydrogenase in relation to serum interleukin -1β pg/ml. Pearson correlation coefficient: r = 0.320* (P = 0.054)|
Click here to view
| Discussion|| |
The results showed that there was negative correlation between SpO2 percentage and IL-6 level (P = 0.002; R = 0.372). This suggests that IL-6 has a direct effect on SpO2 percentage in severe COVID-19 patients, as the severity of the disease was linked to a decrease in SpO2, which could be linked to higher IL-6 levels. These results are similar to the previous study reported by Wu et al. who found the level of IL-6 was negatively correlated with the level of blood oxygen saturation. Moreover, Kadhim showed a significant increase in the serum level of IL-6 in COVID-19 patients, and these increased was effectively associated with the diseases severity in the patients. These findings suggest that IL-6 levels appear to be linked to respiratory failure. SARS-CoV-2 stimulates both innate and adaptive immune responses, causing IL-6 and other cytokines to be produced as well as increased lung capillary permeability, causing ARDS, and accelerated the coagulation pathway, causing microthrombi in lung circulation and increasing the risk of thrombotic events. The fact that IL-6 is primarily produced by wounded lungs could explain the observed connections between cytokine levels and oxygen requirements.
This current study found the increased levels of creatinine was shown to be positively correlated to higher levels of IL-6 (P = 0.012; R = 0.309), as well as the serum albumin level had a negative correlation with increased level of IL-6 and IL-1α (P = 0.034, R = −0.301; P = 0.039; R = −0.259, respectively). This implies that IL-1α has a direct effect on indicate on albumin level. Whereas, the level of IL-6 has a direct impact on both creatinine and albumin levels. These findings are agreed with Liu who found that creatinine was positively correlated with IL-6 (P = 0.001; R = 0.355). There was also a negative correlation between higher levels of IL-6 and lower levels of serum albumin (P = 0.001; R = −0.467), according to the study. Moreover, Xu refer that the severity of the disease was linked to a decrease of serum albumin, which could be linked to an increase in IL-6 and IL-1α levels.
This explains that dysregulation of immunological responses (cytokine storm) caused by SARS-CoV-2 may impact renal injury in COVID-19 patients. Meanwhile, IL6 causes renal endothelial cells to release proinflammatory cytokines and chemokines, which leads to increased vascular permeability in the kidneys and contributes to microcirculatory dysfunction. Capillary leak syndrome and thrombosis can also be caused by proinflammatory cytokines, which can lead to disseminated intravascular coagulation. Furthermore, large amounts of circulating cytokines can cause cell death and tissue damage, resulting to multiorgan failure in kidneys (disturbances of vascular hemostasis, and cytokine-induced injuries).
The present study found the elevation in ALP level has a positive correlation with an increased in IL-1α level (P = 0.024; R = 0.304). While, elevated AST levels were found with be positively correlated to increased IL-1β level (P = 0.049; R = 0.255). Which indicate there was a direct effect of IL-1α on the level of ALP and direct impact of IL-1β on the AST level. These results are consistent with Portincasa who reported that during the progression of COVID-19, the liver could be a primary target of SARS-CoV-2 as well as a secondary target of the virus's complicated systemic changes pathways, which include inflammation and cytokine production (including IL-1, IL-6, and IL-10), immunological response, altered coagulation, hepatic ischemia and hypoxia, and sepsis-related abnormalities.
This explains that patients with more severe disease are more prone to develop liver damage, which is accompanied by changes in liver function tests. Ischemic/hypoxic liver injury, as well as immunologic, inflammatory, and toxic processes induced by systemic sepsis, is aggravating factors. Although viral inclusions do not appear to be present in the liver, this possibility deserves further examination due to the possibility of viral RNA translocation from the gut through portal circulation.
The present study showed that there was positive correlation between the IL-1β level and level of LDH (P = 0.054; R = 0.320). These results are disagreed with Liu who mentioned that there was no significant correlation between LDH level and IL-1β (P = 255; R = −0.065). Because it is linked to metabolic acidosis, in COVID-19 disease, LDH is a powerful predictor of cytokine storm. When pyruvate is converted to lactic acid and accumulates in viral infections, tissue hypoxia prevails. Lactic acidosis causes monocytes and macrophages to generate IL-1, which activates inflammatory responses by triggering the inflammasome. The LDH levels increased as a homeostatic response to acidosis. As a result, LDH is a measure of tissue damage induced by viremia and a dysregulated immune response since LDH elevates as tissue degradation develops. It can predict the progression or regression of disease since it reflects the degree of several pathophysiological processes.
| Conclusions|| |
This study further confirms the growing evidence on the direct role of proinflammatory cytokines in the biochemical changes showed in COVID-19.
We appreciate the efforts of the laboratory team of Al-Amal Hospital in AL-Najaf/Iraq, in particular (Nasser Adrees).
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Ahmadian E, Hosseiniyan Khatibi SM, Razi Soofiyani S, Abediazar S, Shoja MM, Ardalan M, et al
. COVID-19 and kidney injury: Pathophysiology and molecular mechanisms. Rev Med Virol 2021;31:1-13.
Cruz AS, Mendes-Frias A, Oliveira AI, Dias L, Castro AG, Silvestre R. Interleukin-6 Is a biomarker for the development of fatal severe acute respiratory syndrome coronavirus 2 pneumonia. Front Immunol 2021;12:1-10.
Ghaith MM, Albanghali MA, Aldairi AF, Iqbal MS, Almaimani RA, AlQuthami K, et al.
“Potential Predictors of Poor Prognosis among Severe COVID-19 Patients: A Single-Center Study”, Canadian Journal of Infectious Diseases and Medical Microbiology 2021;2021:Article ID 6656092.
Ghazavi A, Ganji A, Keshavarzian N, Rabiemajd S, Mosayebi G. Cytokine profile and disease severity in patients with COVID-19. Cytokine 2021;137:155323.
Letelier P, Encina N, Morales P, Riffo A, Silva H, Riquelme I, et al.
Role of biochemical markers in the monitoring of COVID-19 patients. J Med Biochem 2021;40:115-28.
Liu QQ, Cheng A, Wang Y, Li H, Hu L, Zhao X, et al.
Cytokines and their relationship with the severity and prognosis of coronavirus disease 2019 (COVID-19): A retrospective cohort study. BMJ Open 2020;10:e041471.
Mahase E. China coronavirus: What do we know so far? BMJ 2020;368:m308.
Portincasa P, Krawczyk M, Machill A, Lammert F, Di Ciaula A. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID- 19. The COVID-19 resource centre is hosted on Elsevier connect, the company' s public news and information. Eur J Intern Med 2020;77:19-24.
Guirao JJ, Cabrera CM, Jiménez N, Rincón L, Urra JM. High serum IL-6 values increase the risk of mortality and the severity of pneumonia in patients diagnosed with COVID-19. Molecular Immunology, 2020;128: 64-68.
Sohrabi C, Alsafi Z, O'Neill N, Khan M, Kerwan A, Al-Jabir A, et al
. World Health Organization declares global emergency: A review of the 2019 novel coronavirus (COVID-19). Int J Surg 2020;76:71-6.
WHO (2021). Clinical management Living guidance COVID-19. World Health Organization, 2021.
Wu J, Shen J, Han Y, Qiao Q, Dai W, He B, et al
. Upregulated IL-6 indicates a poor COVID-19 prognosis: A call for tocilizumab and convalescent plasma treatment. Front Immunol 2021;12:1-14.
Xu W, Huang C, Fei L, Li Q, Chen L. Dynamic changes in liver function tests and their correlation with illness severity and mortality in patients with COVID-19: A retrospective cohort study. Clin Interv Aging 2021;16:675-85.
Chen N, Zhou M, Dong X, Qu J, Gong F, Han Y, et al
. Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China: A descriptive study. Lancet 2020;395:507-13.
Ali Saad K, Younus Jasim A. Serum levels of interleukin 6, ferritin, C reactive protein, lactate dehydrogenase, D dimer, and count of lymphocytes and neutrophils in COVID 19 patients: Its correlation to the disease severity. Biomed Biotechnol Res J 2021;5:69.
Verma T, Manish Sinha, Bansal Nitin, Shyam Raj Yadav, Kamal Shah, Nagendra Singh Chauhan. A review on Coronavirus Disease and potentially active drugs targeting coronavirus. Biomed Biotechnol Res J 2021;5:110. [Full text]
[Figure 1], [Figure 2]
[Table 1], [Table 2]