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 Table of Contents  
ORIGINAL ARTICLE
Year : 2021  |  Volume : 21  |  Issue : 4  |  Page : 167-173

Role of microRNA as a marker in detection of diabetic nephropathy in type-2 diabetic Egyptian patients


1 Department of Internal Medicine and Nephrology, Faculty of Medicine, Menoufia University, Menoufia, Egypt
2 Department of Medical Biochemistry and Molecular Biology, Faculty of Medicine, Menoufia University, Menoufia, Egypt
3 Department of Internal Medicine, Faculty of Medicine, Menoufia University, Menoufia, Egypt

Date of Submission25-Jun-2021
Date of Acceptance29-Jul-2021
Date of Web Publication27-Oct-2021

Correspondence Address:
Dr. Elsayed Ghonamy
Department of Internal Medicine, Faculty of Medicine, Menoufia University, Tahlet Burden, Zagazig City Sharkia Government, 44523
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jesnt.jesnt_17_21

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  Abstract 


Background Diabetic kidney disease (DKD) represents the major cause of chronic kidney disease. New biomarkers for early diagnosis of DKD are of interest. Our objective is to elucidate the significance of microRNA-192 (miRNA-192) and neutrophil gelatinase-associated lipocalin (NGAL) as early predictors of diabetic nephropathy in comparison with albuminuria in type-2 diabetic patients.
Patients and methods A number of 45 persons in three groups, group 1: 15 healthy persons with no diabetes mellitus (DM). Group 2: 15 patients diagnosed with type-2 diabetes mellitus (T2DM) without albuminuria. Group 3: 15 patients diagnosed with T2DM with albuminuria. Estimation of serum miRNA, NGAL, urine albumin/creatinine ratio, serum creatinine, urea, fasting blood glucose, 2-h postprandial blood glucose, and glycated hemoglobin.
Results The study showed that the serum miRNA-192 and NGAL levels were significantly higher in patients diagnosed with T2DM with albuminuria in comparison with patients diagnosed with T2DM without albuminuria. There was a significant positive correlation between miRNA-192 and NGAL and urine albumin/creatinine ratio.
Conclusion Serum NGAL and miRNA-192 may serve as tools to follow up the development and progression of DKD.

Keywords: albuminuria, diabetic kidney disease, microRNA-192


How to cite this article:
Khamis SS, Yassin YS, Tawfeek AR, Kasem HE, Ibrahim SM, Ghonamy E. Role of microRNA as a marker in detection of diabetic nephropathy in type-2 diabetic Egyptian patients. J Egypt Soc Nephrol Transplant 2021;21:167-73

How to cite this URL:
Khamis SS, Yassin YS, Tawfeek AR, Kasem HE, Ibrahim SM, Ghonamy E. Role of microRNA as a marker in detection of diabetic nephropathy in type-2 diabetic Egyptian patients. J Egypt Soc Nephrol Transplant [serial online] 2021 [cited 2022 Aug 16];21:167-73. Available from: http://www.jesnt.eg.net/text.asp?2021/21/4/167/329321




  Introduction Top


Chronic kidney disease (CKD) is a major health problem because of its association with a high cardiovascular risk. Most CKD patients progress to end-stage renal disease (ESRD) and require dialysis or transplantation. Type-2 diabetes mellitus (T2DM) is one of the most important risk factors in the development of CKD as around 30–50% of ESRD patients worldwide come from a diabetic origin [1]. Microalbuminuria may not be a sensitive and specific predictor for diabetic kidney disease (DKD) as only ∼30% of patients with microalbuminuria develop overt nephropathy after 10 years of follow-up [2]. Approximately 30–45% of T2DM patients with nephropathy did not have an increase in urine albumin/creatinine ratio defined as normoalbuminuric DKD [3]. One important problem is the growing number of diabetic kidney-disease patients, which highlights the urgent need for novel biomarkers that allow an earlier diagnosis of renal damage [4].

MicroRNAs (miRNAs) are endogenous noncoding single-stranded RNA transcripts most frequently of 19–25 nucleotides in length. It acts as posttranscriptional regulators of gene expression. The identification of differences in miRNA expression may still provide important information on disease mechanism. miRNAs are highly enriched in the endothelial cells. They contribute to the maintenance and repair of vascular integrity and angiogenesis [5].

Neutrophil gelatinase-associated lipocalin (NGAL) is a 25-kDa molecule that belongs to the superfamily of lipocalin proteins. It was initially identified in neutrophils. NGAL is a large glycosylated protein synthesized in renal tubular epithelial cells that is related to antibacterial properties or cellular apoptosis [6].

Currently, clinical studies have highlighted the role of NGAL for detecting diabetic nephropathy [3].


  Patients and methods Top


The present study was carried out on a number of 45 persons from Suez Hospital of Insurance Egypt (who are in the age group of 30–60 years). All were given informed consent and the study was approved by the local medical committee of the hospital.

They were divided into three groups:
  • Group 1: 15 healthy persons with no DM, as well as age, sex, and ethnic origin matched to the patients.
  • Group 2: 15 patients diagnosed with T2DM without albuminuria.
  • Group 3: 15 patients with diabetic nephropathy, assessed by using albumin-to-creatinine ratio in spot urine sample.


Inclusion criteria: T2DM patients above 30 years old.

Exclusion criteria: patients below 30 years, pregnancy, malignancy, patients on angiotensin-converting enzyme inhibitors (ACEIS) or angiotensin II receptor blockers (ARBS) or sodium-glucose cotransporter-2 (SGLT2) inhibitors.

All participants were subjected to

Detailed medical history, physical examination, and measurement of fasting blood glucose, 2-h postprandial blood glucose and glycated hemoglobin (HbA1c), urine albumin-to-creatinine ratio in spot urine sample, serum urea, creatinine, serum miRNA-192, and NGAL.

Statistical analysis

Data were analyzed using Statistical Program for Social Science (SPSS), version 20 (SPSS Statistics is a software package used for logical batched and non-batched statistical analysis). Quantitative data were expressed as mean±SD. Qualitative data were expressed as frequency and percentage.

The following tests were done: independent samples t test of significance was used when comparing between two means, χ2 test of significance was used in order to compare proportions between two qualitative parameters, and Pearson’s correlation coefficient (r) test was used for correlating data, P value.

P value less than or equal to 0.05 was considered significant.

P value less than or equal to 0.001 was considered highly significant.

P value more than 0.05 was considered insignificant.

The sample sizing assumes the expected mean difference for serum miRNA-155 values between controls and type-2 diabetic patients with microalbuminuria to be 0.72-ΔΔCt [7]. To achieve 90% power to detect this difference with a significance level of 5%, it is estimated that 11 participants would be required. With a withdrawal/nonevaluable patient rate of 10%, a total of 13 per-group patients will be recruited, leading to a total required sample size of 39 patients for three groups (control, type-2 diabetic patients with normoalbuminuria, and type-2 diabetic patients with microalbuminuria).


  Results Top


There were no statistically significant differences among demographic data among all studied groups ([Table 1]). There was no significant difference in DM duration between diabetic patients with normoalbuminuria and those with microalbuminuria, P=0.0739 ([Table 2]). There was no significant difference in mean HbA1c between diabetic patients with normoalbuminuria and those with microalbuminuria, P=0.189. The mean HbA1c was significantly higher in patients with microalbuminuria and those diabetic with normoalbuminuria in comparison with the control group (P<0.001, P=0.001, respectively) ([Table 3]).
Table 1 Comparison between control group, diabetes mellitus without albuminuria group, and diabetes mellitus with albuminuria group as regards demographic data

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Table 2 Comparison between control group, diabetes mellitus without albuminuria group, and diabetes mellitus with albuminuria group as regards diabetes mellitus duration

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Table 3 Comparison between control group, diabetes mellitus without albuminuria group, and diabetes mellitus with albuminuria group as regards glycated hemoglobin, 2-h postprandial, and fasting blood glucose

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No significant difference between the control group and each of those diabetic with microalbuminuria or normoalbuminuria regarding serum creatinine (P=0.821, 0.431, respectively). Also, there was no significant difference between the control group and each of those diabetic with microalbuminuria or normoalbuminuria regarding serum urea (P=0.085, 0.656, respectively) ([Table 4]).
Table 4 Comparison between control group, diabetes mellitus without albuminuria group, and diabetes mellitus with albuminuria group as regards serum urea and creatinine

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There was a statistically significant increase in urine albumin/creatinine ratio in DM with albuminuria group in comparison with both control group and DM without albuminuria group (P=0.001, 0.001, respectively) ([Table 5]).
Table 5 Comparison between control group, diabetes mellitus without albuminuria group, and diabetes mellitus with albuminuria as regards urine albumin/creatinine ratio

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The level of serum miRNA-192 is more significantly elevated in patients with microalbuminuria than those with normoalbuminuria (P=0.000) ([Table 6]). There was a significant positive correlation between miRNA-192 and urine albumin/creatinine ratio in patients with microalbuminuria ([Figure 1]). There was a significant positive correlation between NGAL and urine albumin/creatinine ratio in patients with microalbuminuria ([Figure 2]).
Table 6 Comparison between control group, diabetes mellitus without albuminuria group, and diabetes mellitus with albuminuria group as regards serum neutrophil gelatinase-associated lipocalin and microRNA-192

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Figure 1 Correlation between microRNA and albumin/creatinine ratio regarding DM with albuminuria group (r=0.774 and P=0.001). DM, diabetes mellitus.

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Figure 2 Correlation between NGAL and albumin/creatinine ratio in DM with albuminuria group (r=0.586 and P=0.022). DM, diabetes mellitus; NGAL, neutrophil gelatinase-associated lipocalin.

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There was a statistically significant increase in serum NGAL in DM with albuminuria group in comparison with both control group and DM without albuminuria group (P=0.000, 0.041, respectively) ([Table 6]). There was a positive correlation between NGAL with miRNA in patients with microalbuminuria (r=0.701, P=.0.004) ([Figure 3]).
Figure 3 Correlation between NGAL and microRNA in DM with albuminuria group (r=0.701 and P=0.004). DM, diabetes mellitus; NGAL, neutrophil gelatinase-associated lipocalin.

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  Discussion Top


DKD remains the most common cause of ESRD in the United States and most countries. Although albuminuria is the most widely used test for early detection of DKD, it has multiple drawbacks [8]. This triggers exploring new biomarkers for the identifying early diagnosis and prognosis of DKD.

In the present study, there were no significant differences between all studied groups regarding demographic data, which ensures the comparability of the groups for further comparisons. As regards laboratory data, the lack of difference regarding serum creatinine and urea between both groups may be because they are a late indicator of impairment of kidney function [9].

In the current study, the HbA1c level in the microalbuminuric diabetic group showed nonsignificant difference compared with diabetic normoalbuminuric group but significantly higher in comparison with the control group. These findings are in agreement with Saadi et al. [10], who found the same results and in controversy with Sun et al. [11] who recorded that the HbA1c level in the microalbuminuric diabetic group showed a significant increase compared with diabetic normoalbuminuric group. This could be due to the metabolic memory of DM that cannot be detected by HbA1c level or due to the genetic difference between the patients as the hyperglycemia in diabetics is not the only predisposing factor for DKD development.

In the present study, the miRNA-192 blood level is upregulated in participants with DKD and T2DM patients with normal albuminuria. But its level was significantly higher in participants with DKD compared with T2DM patients with normal albuminuria. Moreover, a significant positive correlation was also found between miRNA-192 and the amount of urinary albumin. These results agreed with Saadi et al. [10] who found a significantly higher level of miRNA-192 blood in patients with diabetic nephropathy than the newly diagnosed patients without microvascular complications. Contrariwise to our results, El-Monem et al. [12] found that miRNA-192 expression was significantly lower in T2DM patients with microalbuminuria than those with normoalbuminuria. Some of the differences may relate to models and/or experimental conditions; however, another explanation is that some effects of miRNAs are likely to be indirect in nature.

In the current study, there was a statistically significant increase in NGAL in DM with albuminuria group than DM without albuminuria group. Also, there was a statistically significant increase in serum NGAL in DM without albuminuria group and DM with albuminuria group in comparison with the control group. This was consistent with Nauta et al. [13] who demonstrated that NGAL was 1.5-fold significantly elevated in diabetic patients without albuminuria compared with nondiabetic control group. In our study, serum NGAL showed a significant positive correlation with urinary albumin/creatinine in DM with albuminuria group, Fu et al. [14] found the same results.On the contrary, Aslanhan et al. [15] found that serum NGAL levels did not show any difference between normoalbuminuric and microalbuminuric patients with T2DM. They also did not find any correlation between NGAL levels and urinary albumin excretion. As NGAL is a smaller protein than albumin, it is presumed that it is rapidly filtered and reabsorbed by proximal tubules in normal circumstances. Only 0.1–0.2% of filtrated NGAL is expected in urine. In comparison with glomerular signs, the increasing levels in NGAL occur earlier and this condition might be secondary to subclinical tubular impairment [16].


  Conclusion Top


From this study, we can conclude that serum NGAL and miRNA-192 may serve as tools to follow-up clinical monitoring of the development and progression of DKD.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

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Donate-Correa J, Luis-Rodríguez D, Martín-Núñez E, Tagua VG, Hernández-Carballo C, Ferri C et al. Inflammatory targets in diabetic nephropathy. J Clin Med 2020; 9:458.  Back to cited text no. 1
    
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Motawi TK, Shehata NI, ElNokeety MM, El-Emady YF. Potential serum biomarkers for early detection of diabetic nephropathy. Diabetes Res Clin Pract 2018; 136:150–158.  Back to cited text no. 2
    
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Kaul A, Behera MR, Rai MK, Mishra P, Bhaduaria DS, Yadav S et al. Neutrophil gelatinase-associated lipocalin: as a predictor of early diabetic nephropathy in type 2 diabetes mellitus. Indian J Nephrol 2018; 28:53–60.  Back to cited text no. 3
    
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American Diabetes Association, Microvascular complications and foot care. Standards of medical care in diabetes-2020. Diabetes Care 2020; 43:S135–SS51.  Back to cited text no. 4
    
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Ortega FJ, Mercader JM, Moreno-Navarrete JM, Rovira O, Guerra E, Esteve E et al. Profiling of circulating microRNAs reveals common micro-RNAs linked to type 2 diabetes that change with insulin sensitization. Diabetes Care 2014; 37:1375–1383.  Back to cited text no. 5
    
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Rahimipour A. Assessment of neutrophil gelatinaseassociated lipocalin (NGAL) as an early biomarker for detection of renal impairment in hypertensive patients. J Paramed Sci 2014; 5:63–66.  Back to cited text no. 6
    
7.
Mohany KMM, Rezk MY, Elkatawy HA. Circulating miR-155 Levels correlate with serum neutrophil gelatinase associated lipocalin in patients with diabetic nephropathy. Int J Diabetes Res 2017; 6:41–46.  Back to cited text no. 7
    
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Chen Y, Lee K, Ni Z, He JC. Diabetic kidney disease: challenges advances, and opportunities. Kidney Dis 2020; 6:215–225.  Back to cited text no. 8
    
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Levey AS, Coresh J, Tighiouart H, Greene T, Inker LA. Strengths and limitations of estimated and measured GFR. Nat Rev Nephrol 2019; 15:785–786.  Back to cited text no. 9
    
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Saadi G, El Meligi A, El-Ansary M, Alkemary A, Ahmed G. Evaluation of microRNA-192 in patients with diabetic nephropathy. Egypt J Internal Med 2019; 31:122–128.  Back to cited text no. 10
    
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Sun YM, Su Y, Li J, Wang LF. Recent advances in understanding the biochemical and molecular mechanism of diabetic nephropathy. Biochem Biophys Res Commun 2013; 433:359–361.  Back to cited text no. 11
    
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El-Monem A, Mahfouz M, Mohamed M. Microrna 192 gene expression in type II diabetic nephropathy. Egypt J Hosp Med 2017; 68:885–893.  Back to cited text no. 12
    
13.
Nauta FL, Boertien WE, Bakker SJ, van Goor H, van Oeveren W, de Jong PE, Bilo H, Gansevoort RT. Glomerular and tubular damage markers are elevated in patients with diabetes. Diabetes Care 2011; 34:975–981.  Back to cited text no. 13
    
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Fu WJ, Xiong SL, Fang YG, Wen S, Chen ML, Deng RT et al. Urinary tubular biomarkers in short-term type 2 diabetes mellitus patients: a cross-sectional study. Endocrine 2013; 41:82–88.  Back to cited text no. 14
    
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Aslanhan E, Ojalvo D, Özsenel EB, Basat SU, Borlu F. Association of neutrophil-gelatinase-associated lipocalin with microvascular complications in patients with type 2 diabetes: a cross-sectional study. Cardiovasc Endocrinol Metab 2019; 8:82–87.  Back to cited text no. 15
    
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Zachwieja J, Soltysiak J, Fichna P, Lipkowska K, Stankiewicz W, Skowronska B et al. Normal-range albuminuria does not exclude nephropathy in diabetic children. Pediatr Nephrol 2010; 25:1445–1451.  Back to cited text no. 16
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]



 

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