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Can depression and anxiety consider as risk factor for expression of RIG-1 and MDA5 in the chronic HBV infected patients?

Research

Content details

Med ID: 194
Read Online: 440
Published Online: 19 , January, 2020
Condition: Accepted
In Subjects: Diseases

Manuscript

Vahid Mohammadi-Shahrokhi, Maryam Ebrahim, Reza Bidaki, Mehdi Khaleghinia, Seyed Mohsen Mousavi, Mohammad Kazemi Arababadi


Abstract

Background: MDA5 and RIG-1, the well known pattern recognition receptors (PRRs), play important roles in viral nucleic acid recognition. Our previous investigation revealed that MDA5 expression was impaired in chronic HBV infected (CHB) patients. Additionally, the prevalence of psychological disorders, which alters immune responses, is high in CHB patients. Therefore, MDA5 and RIG-1 mRNA levels were examined in the peripheral blood immune cells of CHB patients with various degrees of anxiety and depression to evaluate the plausible roles played by the psychiatric disorders on MDA5 and RIG-1 expression.

Methods: In this study, MDA5 and RIG-1 mRNA levels were examined in 60 CHB patients who suffered from various anxiety and depression degrees using the Real-Time PCR technique.

Results: MDA5 and RIG-1 mRNA levels did not significantly alter among CHB patients with various states of anxiety and depression.

Conclusions: According to the results psychiatry disorders are unable to change MDA5 and RIG-1 expression levels. Thus, impaired expressions of MDA5 in CHB patients were not associated with anxiety and depression.

Keywords: Chronic hepatitis B infection, MDA5, RIG-1, Depression, Anxiety.

Introduction

Chronic hepatitis B virus (HBV) infected (CHB) patients exhibit an impaired specific response to HBV, which leads to a chronic clinical form in which HBV do not completely clear from both hepatocytes and serum (1). Based on the chronic inflammation, which is produced in responses to HBV, CHB could be considered as a significant cause of liver complications including cirrhosis and hepatocellular carcinoma (HCC) (2, 3). Although, our knowledge regarding the mechanisms responsible for prolonged hepatitis B infections are growing in recent years, the main environmental factors which may confer immune responses are yet to be illuminated. It has been demonstrated that intracellular pattern recognition receptors (PRRs) may play significant roles against viral infections (4, 5). Melanoma differentiation-associated protein 5 (MDA5) and retinoic acid-inducible gene 1 (RIG-1) are two key intracellular PRRs which are able to induction of several intracellular signaling pathways against viral dsRNA (6, 7). Our previous investigation demonstrated that CHB patients suffered from altered expression of MDA5 and also RIG-1 (8). Accordingly, we have reported that MDA5 and RIG-1 expression were decreased and increased, respectively, in the peripheral blood immune cells of the patients (8). Both increasing and decreasing PRRs may be considered as a risk factor for the pathogenesis of CHB and its complications. Investigators are evaluating the main mechanisms which alter expression of immune related molecules in the CHB patients. As mentioned, environmental factors can be considered as risk factors for induction of some alterations in immune cell functions (9). Psychological disorders, as important environmental factors, are able to alter immune responses (10, 11). Thus, anxiety and depression, as psychological disorders, may be the main parts of CHB altered immune responses puzzle. Therefore, this study was aimed to determine the effect of anxiety and depression on the MDA5 and RIG-1 mRNA levels in CHB patients.

Material and methods

Subjects: The participants, CHB patients, in this study were exactly the subjects who were under evaluation of MDA5 and RIG-1 expression in our previous study (8). Accordingly, 60 CHB patients were evaluated regarding the statuses of anxiety and depression using a standard questionnaire. The methods for confirmation of CHB, excluding and including criteria and collections of peripheral blood samples were described previously (8). In order to confirm the HBV infection, HBV markers were evaluated in all the participants. The diagnosis of anxiety and depression was performed by an expert psychologist, according to the fill lay out of standard Hamilton and Beck questionnaires. Accordingly, the results obtained from questionnaire were reviewed and patients were selected by an expert psychologist. The project protocol was approved by the Ethical Committee of Rafsanjan University of Medical Sciences by IR.RUMS.REC.1394.128 code number.

Evaluation of HBV markers: All of the samples were tested regarding hepatitis B surface antigen (HBsAg) and HBV-DNA using enzyme linked-immunosorbent assay (Behring, Marburg, Germany) and Real-Time PCR (Design primer, London, UK), respectively, according to the manufacture’s guidelines.

MDA5 and RIG-1 mRNA levels detection

RNA purification/cDNA synthesize/Real-time PCR protocols were defined previously (12). Briefly, total RNA was purified using a commercial kit from the Sinaclon Company (Tehran, Iran), cDNA was synthesized by a kit from the Parstous Company (Tehran, Iran) and the cDNA was amplified in parallel with β-actin using a kit from Parstous (Tehran, Iran) in Bio-RAD CFX96 (Bio-RAD Com, Washington, USA) instrument. The primer sequences were as follow:

MDA5 forward: GCAGAGGTGAAGGAGCAGA, MDA5 reverse: AAACGATGGAGAGGGCAAG, RIG-1 forward: CACACCAAGAGCCCAAAC, RIG-1 reverse: TGACCCGATAGCAACAGC, β-actin forward: GGCACCCAGCACAATGAAG, β-actin reverse: CCGATCCACACGGAGTACTTG.

Data analysis

The differences regarding MDA5 and RIG-1 mRNA levels among various groups were analyzed using One Way Anova under SPSS software version 18. P value > 0.05 considered significant.

Results

Prevalence of anxiety and depression in CHB patients

Anxiety and depression prevalence in CHB patients was reported in our previous investigation (13). Accordingly, 60, 11.6, 11.6 and 16.8 percent of the CHB patients were associated with no, mild, moderate and severe anxieties, respectively. And 63.3% had no depression, 21.7% had mild depression and 15% had moderate depression. None of the patients suffered from severe depression.

HBV markers

Evaluation of HBV markers revealed that all of the patients had detectable HBsAg and HBV-DNA.

Expression levels of target genes:

The results indicated that MDA5 expression levels in the patients with no, mild and moderate depression were 0.51 ± 0.18, 0.07 ± 0.05, 0.03 ± 0.01, respectively, with no significant differences (p= 0.143). These findings also demonstrated that expression levels of RIG-1 in patients with various degrees of depression were not differed (p= 0.558) (Figure 1). Expression levels of MDA5 (p= 0.271) and RIG-1 (p= 0.385) in patients with various degrees of anxiety were also not differed (Figure 2).

Figure 1. MDA5 and RIG-1 mRNA levels in CHB patients suffering from no, mild and moderate depression. The figure demonstrates of MDA5 and RIG-1 expression levels did not differ among various states of depressed patients.
Figure 2. Relative MDA5 and RIG-1 expression levels in CHB patients suffering from no, mild, moderate and severe anxiety. The figure illustrates that MDA5 and RIG-1 mRNA levels did not differ among patients.

Discussion

Our previous investigations demonstrated MDA5 and RIG-1 expression levels significantly decreased in the CHB patients (8). The results from the current study revealed that neither anxiety nor depression affects expression of MDA5 and RIG-1 in the CHB patients (Figure 1 and 2). Based on the results it seems that anxiety and depression, as psychiatric disorders are unable to alter MDA5 and RIG-1 expressions. Thus, it may be concluded that psychiatric disorders may not be considered as the risk factors for down or up-regulation of MDA5 and RIG-1, as important intracellular PRRs. However, based on the fact that more than 40% of the CHB patients suffered from anxiety and depression, it appears that more investigations regarding the effects of the psychiatric disorders on the expression of other immune related molecules in the CHB patients. In parallel with the current results, our previous study showed that anxiety and depression were also unable to alter expression of interleukin-1 receptor associated kinases 4 (IRAK4), TNF receptor associated factor 3 (TRAF3) and interferon regulatory factor 7 (IRF7) in CHB patients (13). IRAK4 and TRAF3 are the important intracellular molecules which participate in the intracellular signaling pathways of toll like receptors (TLRs), the most important innate receptors (14-16). IRF7 also is a transcription factor which not only is activated by MDA5 and RIG-1 related pathways but also is the target of TLR pathways (8, 17, 18). However, another study from our research team revealed that anxiety is an important environmental factor which leads to CD36, a scavenger receptor on the surface of monocytes/macrophages, up-regulation (19). The effects of depression on the serum levels of transforming growth factor-β (TGF-β) CHB patients have also been documented by Bahramabadi and colleagues (20).

Based on the results, it may be hypothesized that intracellular sensors like MDA5 and RIG-1 and their related molecules, such as IRF7, may not be affected by psychiatric disorders and may cell surface receptors, including CD36 and related cytokines such as TGF-β may be the target of the disease to alter immune responses.

However, based on the fact that several molecules including receptor interacting protein 1 (RIP1) and IFN-β promoter stimulator-1 (IPS-1), also participate in the MDA5 and RIG-1 signaling pathways, it appears that more investigations need to be designed to fill out the puzzles of the effects of psychiatric disorders on the expression of MDA5 and RIG-1 pathways.

Acknowledgements:

This project was supported by a grant from the Rafsanjan University of Medical Sciences.

Conflict of interest:

Authors have no conflict of interest to declare.

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