• Nu S-Au Găsit Rezultate

View of Association between Interleukin-18 (-607 C→A) Polymorphisms and Risk of Chronic Kidney Disease

N/A
N/A
Protected

Academic year: 2022

Share "View of Association between Interleukin-18 (-607 C→A) Polymorphisms and Risk of Chronic Kidney Disease"

Copied!
8
0
0

Text complet

(1)

Association between Interleukin-18 (-607 C→A) Polymorphisms and Risk of Chronic Kidney Disease

Manar Hussein AL-Aboudy1 *, Musa Nima AL- Jiafry2, Mahdi Hussein Al-Ammar3

1* M.Sc, Corresponding author, Department of Pharmacy/ College of Pharmacy/ The Islamic University;

Najaf- Iraq

E-mail [email protected]

2Professor, Department of Biology / College of Science / University of Kufa 3 Professor, Department of Biology / College of Science / University of Kufa

Abstract

Background: We designed this study to determine the association between gene polymorphism of IL-18 (-607 C→A) with the development of chronic kidney disease using ARMS-PCR. The present study detected IL-18 -607 C→Agenotype frequency distribution in CKD patients and control.

Methods: A total number of 113 clinical samples were collected from patients who suffer from chronic kidney disease (CKD) .The complete genomic DNA of 113 patients with CKD and 100 of control was extracted employing a traditional kit of the entire human genomic DNA extraction kit.

Results : The genotypes relative frequency in CKD patients were as follows. CC (1.76%), CA (98.2%) and AA (0%). C allele has a close frequency in CKD patients with (50.4%) to A allele (49.5%). There was no significant difference (P value = 0.49) among CKD patients and control group. Data of the current study in chronic kidney disease patients showed that CA heterozygote genotype was the most frequent, followed by CC homozygote while there is no AA allele in this study. However, There was no significant differences in the IL-18 allele frequency distribution between control and chronic kidney disease patients in which (P value = 0.49).Conclusions : This study indicate that IL-18 - 607 C→A allele do not consider a risk factor for effection on the level of IL-18 in patients with chronic kidney disease.

Keywords: Chronic kidney disease, IL-18, Polymorphisms, ARMS-PCR Introduction

IL-18 cytokine is participated in the regulation of innate and acquired immune responses and has a major role in autoimmune diseases by controlling Th1- and Th2- immune response [1]. The human IL-18 gene contains several polymorphisms in the promoter region [2]. One of these SNP is _607 C/A (rs1946518) which may have the ability to affect the binding of transcription factors and therefore modulate the expression of IL-18 mRNA [3]

ARMS PCR is a flexible, rapid and economical tool of SNP detection [4]. it is a reliable method for the analysis of point mutations or small deletions [5]. ARMS was initially developed for allele-specific amplification without the use of restriction enzymes and radioisotopes, it allows the detection of specific genotypes in a single PCR step followed by gel electrophoresis without the requirement for expensive and frequently difficult post-PCR manipulation [6].

(2)

In the view of the above mentioned introduction, we designed this study to determine the association between gene polymorphism of IL-18 (-607 C→A) with the development of chronic kidney disease using ARMS-PCR.

Materials and Method Collection of samples

A total number of 113 clinical samples were collected from patients who suffer from chronic kidney disease (CKD) at age range (15-80 years) and 100 healthy persons as controls at age range (25–65) years. All subjects were admitted to one of the biggest Hospital including Al-Sadr Medical City in Al-Najaf City/Iraq within two months started in January 2020. The patients in this study included 62 males and 51 females while healthy controls was 50 males and 50 females. whole blood were collected from each participant was used to extract DNA for detection of single nucleotide polymorphisms (SNP) of interleukins 18 (-607 C→A) with the use of amplification refractory mutation system polymorphism PCR (ARMS – PCR) technique and the data were statistically analysed by software SPSS version 24.

DNA extraction and PCR assay

The complete genomic DNA of 113 patients with CKD and 100 of control was extracted employing a traditional kit of the entire human genomic DNA extraction kit ( iNtRoN, Biotech. Inc., Korea), whosesoever, the extraction was performed based on the guidance of manufacture corporation. The nucleic acid was conserved under -20ᵒC state using the deep freezing device, the PCR technique was employed to examine and detect all the genes suggested by Celik. (2018) [7] which described in table (1) . The process of gel document (Cleaver, United Kindom), demanded to check and distribute the migration of PCR bands applying 1% agarose ( iNtRoN, Biotech. Inc., Korea), back dyeing the gel with ethidium bromide at 0.5 μg/ml concentration.

Table ( 1) : Primer sequence and product size

Cytokin e

Primers

Sequence (5ʹ-3ʹ )

Product size ( bp)

IL-18 −607 C→A F primer (C

allele )

GTTGCAGAAAGTGTAAAAATTATTA C

F primer (A 196 allele)

GTTGCAGAAAGTGTAAAAATTATTA A

R primer

TAACCTCATTCAGGACTTCC

(3)

The SPSS V.24 program was used in statistical analysis of the data. Chi- Square was used to compare and extract duplicates and percentages

Result

Genetic Polymorphism of IL-18 Gene

The genetic polymorphism of IL-18 gene was determined at one position ; IL-18-607 C

→A which was present with three genotypes (CC, CA and AA) in chronic kidney disease and control. ARMS-PCR method was used to genotyping of IL-18 -607 C →A in CKD and control. The PCR product size was 196 bp. The positive amplification of both PCR reaction exhibited a CA genotyping, while positive amplification in only the first PCR reaction exhibited a CC genotype.

The results of ARMS-PCR for IL-18-607 C →A among chronic kidney disease patients showed 13 bands of CA heterozygote allele as shown in figure (1).

Figure (1): Agarose gel electrophoresis image that shows the ARMS-PCR product analysis of IL-18 (-607 C→A) in CKD patients . ARMS-PCR product was analyzed by

1% agarose gel. Where M: marker (100bp – 1500bp ). and lane CA heterozygote was shown at the following bands ( 1,2,3,4,5,6,7,8,9,10,11,12,13) digested at ( 196 bp C

allele and 196 bp A allele)

The results of ARMS-PCR for IL-18-607 C →A in healthy control subjects showed 15 bands of CA heterozygote allele as shown in figure (2).

(4)

Figure (2): Agarose gel electrophoresis image that shows the ARMS-PCR product analysis of IL-18 (-607 C→A) in control subjects . ARMS-PCR product was analyzed by 1% agarose gel. Where M: marker (100bp – 1500bp ). and lane CA heterozygote was shown at the following bands (1,2,3,4,5,6,7,8,9,10,11,12,13,14,15)

digested at ( 196 bp C allele and 196 bp A allele).

IL-18 genotype frequency distribution in chronic kidney disease patients and control groups is shown in Table (2). The genotypes relative frequency in chronic kidney disease infection (CKD) were as follow: CC (1.76%), CA (98.2%) and AA (0%).While in control subjects was CC (2%), CA (98%) and AA(0%). There was no significant difference (Chi-square = 1.40 ; P value = 0.49) among CKD patients and control group according to IL-18 genotype detection and distribution as shown in figure (2)

C allele is in has a close frequency in CKD patients with (50.4%) to A allele (49.5%).

There was no significant differences in the IL-18 allele frequency distribution (Chi- square = 0.05 ; P value = 0.97) between control and patients groups , as shown in Table (2) .

Table (2). IL-18 genotype frequency distribution in CKD patients and control groups.

SNP Genotype CKD patients Control Chi Square

P value

CC 2

(1.76%)

2 (2%)

CA 111 98

(5)

-607 C→A (0%) (0%) NS

Total 113

(100%)

100 (100%) Allele frequencies

C 113

(50.4%)

100 (50.5%)

0.05 0.97 NS

A 111

(49.5%)

98 (49.4%)

Total 224

(100%)

198 (100%) NS : Non-significant

Genotype Distribution According to Gender

IL-18 genotyping frequency distribution according genderin chronic kidney disease patients and control groups is shown in Table (3). The genotypes relative frequency in female with CKD were as follows: CC (3.6 %), CA ( 96.3%) and AA ( 0%) , while in control subjects : CC (3.92 %) , CA (96%) and AA 0 (0%). There was no significant difference (Chi-square = 2.68 ; P value = 0.26) among female in CKD patients and control.

While in male with CKD were as follows: CC and AA ( 0% ) , CA ( 100 %) while in control subjects : CC and AA ( 0 %) and CA (100%). There was no significant difference (Chi-square = 2.28 ; P value = 0.32) among male in CKD patients and control.

There was no significant difference (Chi-square = 1.45 ; P value = 0.22) among male and female in CKD patients as well as no significant difference (Chi-square = 1.89 ; P value

= 0.16) among male and female in control .

Table (3). IL-18 Genotype frequency distribution in CKD patients and control groups according gender

IL-18 SNP Gender groups Chi Square

P value

Female Male

CKD patients CC 2 (3.6%) 0 (0%)

1.45 0.22 NS CA 53 (96.3%) 58(100%)

AA 0 (0%) 0 (0%)

Control CC 2 (3.92%) 0 (0%)

1.89 0.16 NS

CA 49(96%) 49 (100%)

AA 0 (0%) 0 (0%)

(6)

Chi Square P value

2.68 0.26 NS

2.28 0.32 NS NS : Non-significant

Discussion

Interleukin-18 (IL-18) belongs to the IL-1 cytokine superfamily with pro-inflammatory features, it serves as a stimulator for the production of interferon c (IFNc)[8]. Many cells like Kupffer cells, monocytes, macrophages, and dendritic cells can produce IL-18. It functions as a modifier for the immune response by inducing gene expression of cytokine, helper T cell differentiation, natural killer cell activations, and also function as a main proinflammatory cytokine in autoimmune and inflammatory diseases [9].

IL-18 is involved in both T helper (Th)1 andTh2 immune responses, due to these multiple functions, IL- 18 is believed to have an important role in host defense toward viral infection [10].IL-18 (_607 C/A) polymorphisms are widely SNPs in the promoter region which have an effect on the IL-18 gene transcription activity and transform its expression and production by disrupting the nuclear factor binding sites for the cAMP- responsive element-binding protein and histone H4 transcription factor (H4TF-1) respectively [11].

Data of the current study in chronic kidney disease patients showed that CA heterozygote genotype was the most frequent, followed by CC homozygote while there is no AA allele in this study .Also there was no significant difference in these genotypes between CKD patients and control. As well as there is no significance difference in the IL-18 allele frequency distribution between control and patients groups.

Data of the current study in both chronic kidney disease similar to Farias et al., (2013) [12] which found that association of the polymorphisms of the IL-18 gene was observed with neither rheumatoid arthritis nor risk factors forcardiovascular disease but unsimilar to Fatima et al., (2017) [13] which detected that the percentage of IL-18 -607 A/A polymorphism was higher in overweight and obese subjects vs normal weight subjects as well as to Umare et al., (2019) [14] which revealed that interleukin-18 polymorphisms elucidated in their study appear to confer genetic susceptibility to the disease and are associated with renal, serositis and neurologic involvement in Indian systemic lupus erythematosus patients.

According to these results and similarity between patients and control groups we cannot consider CA genotype as risk factor for developing chronic kidney failure. Testing for IL- 18 607 C-A revealed that CKD patients showed insignificant variation in the distribution of IL-18 -607 genotypes. Comparing patients with controls indicated that IL-18-607 alleles or genotypes showed no association with the risk of UTI and CKD development in Iraqi population or protection against them

(7)

Conclusion

This study indicate that IL-18 -607 C→A allele do not consider a risk factor for effect on the level of IL-18 in patients with chronic kidney disease.

References

[1] Salimi, E., Karimi-Zarchi, M., Dastgheib, S.A., Abbasi, H., Tabatabaiee, R.S., Hadadan, A..(2019). Association of promoter region polymorphisms of IL-6 and IL- 18 genes with risk of recurrent pregnancy loss: a systematic review and meta- analysis. Fetal Pediatr Pathol Taylor and Francis Ltd, 39:346–59.

[2] Zhang, M-J., Zhou, Y., Wang, X., Chen, X., Pi, Y., Guo, L..(2016). Interleukin-18 gene promoter 607A polymorphism, but not 137C polymorphism, is a protective factor for ischemic stroke in the Chinese population: a meta-analysis. Meta Gene Elsevier, 9:165–72.

[3] Kim, H.L., Cho, S.O., Kim, S.Y., Kim, S.H., Chung, W.S., Chung, S.H. 2012).

Association of interleukin-18 gene polymorphism with body mass index in women.

Reprod Biol Endocrinol, 10:31.

[4] Alyethodi, R.R., Singh, U., Kumar, S., Deb, R., Alex, R., Sharma, S., Prakash, B.(2016). Development of a fast and economical genotyping protocol for bovine leukocyte adhesion deficiency (BLAD) in cattle. SpringerPlus, 5(1):1442

[5] Medrano, R.F.V., de Oliveira, C.A., Medrano, R.F.V., de Oliveira ,C.A.(2014).

Guidelines for the Tetra-Primer ARMS-PCR Technique Development, Mol Biotechnol , 56(7):599-608.

[6] Newton, C. R., Graham, A., Heptinstall, L. E., Powell, S. J., Summers, C., Kalsheker, N (1989). Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS), Nucleic Acids Research, 17(7), 2503–2516.

[7] Celik, S.D.(2018). Genetic analysis of interleukin 18 gene polymorphisms in alopecia areata, J Clin Lab Anal. 32:e22386.

[8] Yue, M., Wang, J.J., Tang, S.D., Feng, L., Zhang, Y., Liu, Y (2013). Association of interleukin-18 gene polymorphisms with the outcomes of hepatitis C virus infection in high-risk Chinese Han population, Immunol Lett, 154: 54–60.

[9] Sedimbi, S., Hägglöf, T., Karlsson, M.I.(2013). IL-18 in inflammatory and autoimmune disease, Cell Mol Life Sci, 70. 4795-08.

[10] Gracie, J.A., Robertson, S.E., McInnes, I. B. (2003). Interleukin-18, J Leukoc Biol, 73:213–24.

[11] Celik, K.S., Öz, Z.S., Dursun, A., Unal, A., Emre, U., Cicek, S..(2014). Interleukin 18 gene polymorphism is a risk factor for multiple sclerosis. Mol Biol Rep, 41:1653–8.

[12] Farias, T.D.J., do Canto, L.M., Medeiros, M,D., Sereia, A.F.R. de Carlos Back, L.K.F., de Mello, F.M., Zimmermann, A.F., Pereira, I.V., Muniz, Y.C., Marrero, A.R. et al. (2013). Ausência de associaçمo entre os polimorfi smos do gene interleucina-18 e artrite reumatoide , Revbrasreumatol , 53(2): 199 – 205.

(8)

[13] Fatima, S.S., Jamil,Z., Abidi,S.H., Nadeem,D., Bashir, Z.; Ansari, A.(2017).

Interleukin-18 polymorphism as an inflammatory index in metabolic syndrome: A preliminary study, World J Diabetes, 8(6): 304-310.

[14] Umare, V., Pradhan, V., Nath, S., Rajadhyaksha, A., Ghosh, K., Nadkarni, A.H.(2019). Impact of functional IL-18 polymorphisms on genetic predisposition and diverse clinical manifestations of the disease in Indian SLE patients.0: 1–10.

[15] Shi, J-H., Niu, L-D., Chen, X-Y., Hou, J-Y., Yang, P., Li, G-P.(2015). Investigation on the IL-18 - 607A/C and -137C/G on the susceptibility of ischemic stroke, Pak J Med Sci 31(1).198-02.

[16] Ma, J.B., Chen, L., Gao, B., Xu, J.(2016). Effect of polymorphisms in interleukin-18 gene on the susceptibility to coronary artery disease in a Chinese population, Genet Mol Res, 15.

Referințe

DOCUMENTE SIMILARE

Confidence level was set at 95% (P value ≤0.05), calculation for the relative risk of developing the disease for specific genotypes was made as following ((the percentage of allele

As expected, the presence of chronic renal lesions (glomerulosclerosis, interstitial fibrosis, arteriolohyalinosis) was associated with a decrease in kidney length, but

The disease is observed in all patients with clinical and laboratory manifestations of kidney inflammation, renal echographic changes are noted, which can be divided

He reviewed the methodology for the statistical analysis of censored survival data which arise from a model in which the factors under investigation act

24 In the present study, we also found that IL-18 607 C/A and 137 G/C genotypes had no effects on any of the studied data, that may be explained by the demonstrated

This paper examines the significance of exploring Alzheimer's Disease utilizing AI, the need to utilize both conduct and natural markers information, and a computational strategy

However, the statistical analysis showed no significant(P>0.05) differences between them. A significant P<0.05) improvement was observed in active sperm motility

Low serum albumin levels are strongly associated with cardiovascular disease, heart failure and mortality especially in elderly population.[16–19] Albumin levels are not present