الفهرس 
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Abstract
Diabetes mellitus is a group of metabolic disorders characterized by chronic hyperglycemia., causing many complications as nephropathy, retinopathy and peripheral neuropathy.
Diabetic nephropathy (DN) is one of the most known microvascular complications of diabetes mellitus (DM), which presents with persistent proteinuria, a progressive decrease in glomerular filtration rate.
The severity of DN is assessed by measuring urinary albumin excretion, such as albumin-to-creatinine ratio (ACR). Microalbuminuria is typically considered as a
marker of DN, while persistent macroalbuminuria is regarded as a predictor of ESRD . However, urinary microalbuminuria, it is not sensitive, because tissue damage and inflammationhave already occurred by the time that microalbuminuria is detectable. Furthermore, microalbuminuria is not specific to DN but is merely a hallmark of glomerular lesions.
Thus, asensitive and reliable biomarkers are needed to evaluate the initiation and development of DN for early intervention.
MicroRNAs (miRNAs), a class of endogenous small non-coding RNAs, are becoming recognized as important regulators in gene expression .MicroRNAs can be remarkably stable in biofluids, and specifically plasma/serum miRNAs have been investigated as biomarkers in several diseases , including diabetic nephropathy.
Change the levels of microRNAs 130b and 146a have an association with diabetic renal damage and development of DN in T2DM patients
The study was done in MINIA university hospital diabetic out-clinic. In current study we included 100 subjects divided into three main groups
group I: prediabetic group , It included 20 patients 4(20%) males and 16(80%) females , their age ranges from (44-62 years.
group II: Diabetic group, It included 60 diabetic patiants 14(23.3%) males and 46(67.7%) females , their age ranges from (40-62 years) , this group divided into three groups according to the level of albuminuria:
• Normoalbuminuria group,( UACR <30 mg/g) It included 20 diabetic patiants,4(20%) males and 16(80%) females.
• Microalbuminuria group, (UACR 30– 300 mg/g) It included 20 diabetic patiants,4(20%) males and 16(80%) females.
• Macroalbuminuria group, (UACR>300 mg/g) It included 20 diabetic patiants with macroalbuminuria , 6(30%) males and 14(70%) females
group III : controlled group, It included 20 healthy volunteers persons. 6 (30%) males and (70%) females , their age ranges from (44-60 years).
The studied groups diagnosed according to American diabetes Association
(ADA) 2017 )We classified them according to FBG, 2hpp BG, and HBA1C.
Patients were subjected to:
1- History and clinical evaluation.
2-Laboratory investigations which included:
A) Glycemic parameters: Fasting Blood Glucose ( FBG), 2 hour post prandial blood glucose (2hppBG), Fasting insulin, HbA1c, HOMA IR, HOMA
β) Lipid profile: (Total Cholesterol (TC),Triglycerides(TG), High Density Lipoprotein Cholesterol (LDLC) and Low Density Lipoprotein Cholesterol (LDLC).
C) Complete Blood Picture .
D) renal functions.
E) Urinary Albumin /Creatinin ratio .
F)Estimated Glomerular Filtration Rate (eGFR)
G) serum levels of microRNAs 130b and 146a detected by PCR.
2-Assessment of symptoms and/or signs using scoring
Patients were also subjected to ultrasound and Fundus examination.
The results of the study showed that: demographic and anthropometric data of the three studied groups (control group, pre-diabetic group and diabetic group) as regard age was ranged in all groups from 40 to 62 yrs, age mean in control group was (50.6±4.4yrs), pre-diabetic group was (53±6yrs) and diabetic group was (53±5.3yrs).
Females were more participated in our study 14(70%) for control group , 16(80%) in pre-diabetics and 46(76.7%) for diabetics. There was no significant difference between the 3 groups as regards systolic and diastolic blood pressure.
Fundus and abdominal ultrasounography results between the three studied groups showed that there was significant increase in the incidence of abnormality of both in diabetic group(p=0.001) compared to other two groups, without significant difference between both (control group and pre-diabetic group).
There were 28 diabetic patients had abnormal fundus examination 6 patients of them had (10%) non proliferative diabetic retinopathy , 12(20%) patients had mild -proliferative diabetic retinopathy,6 patients had (10%) moderate -proliferative diabetic retinopathy , 4(6.7%) patients had severe -proliferative diabetic retinopathy), while all patients in control and pre-diabetic groups had normal fundus examination .
As regard abdominal ultrasounography examination of diabetic patients there was 20 patients (33.3%) had fatty liver and 8 patients (13.3%) kidenys echogenicity grade 1, while in pre-diabetic only 2 patients (10%)had fatty liver.
the diabetic group had significantly higher levels in FBG, 2hour postprandial blood glucose, HBA1c and than pre-diabetic and control group with (p value<0.001).
As regarding HOMA-IR , there was asignificant increase in diabetic and prediabetic groups compared to control group (p=<0.001) , without significant difference between both diabetic and prediabetic groups(p=0.10). However, the diabetic group showed significantly lower levels in HOMA-B than pre diabetics and control (p=<0.001) , without significant difference between both pre-diabetic and control groups (p=0.516).
The comparison of lipid profile between the three studied groups TG, TC, HDL and LDL showed higher significant levels in diabetic and prediabetic groups than control group (p<0.001) with highest levels in diabetic group. , without significant difference between both diabetic and prediabetic groups regarding TG, TC, HDL and LDL with (p value = 0.12 , 0.314 , 0.851 ,0.054 respectively ) .
As regarding serum insulin, there was significant increase in diabetic group compared to control group (p=< 0.001) , and in prediabetic group compared with both diabetic and control groups (p= 0.02 ,0.001 respectivly).
Results of of microRNAs 130b and 146a:
Our study explored the changes of serum miR‐130b levels in T2DM patients. The analysis showed that the serum miR‐130b level of Diabetic group was significantly lower than that of the Pre-diabetic and control groups, With p value (< 0.001) .
Our results showed that serum miR‐130b was obviously negatively correlated with HbA1c levels(r = −0.759, p = 0.001) , FBG level (r=0.709, p < 0.001) and HOMA‐IR(r = −0.603 p < 0.001). Thus, it can be speculated that circulating serum miR‐130b is closely related to blood glucose levels and insulin resistance in patients with T2DM.
In addition, we found that serum miR‐130b exhibited different expression in T2DM patients with different stages of urinary albumin. The levels of circulating serum miR‐130b gradually and markedly decreased and was significantly lower in T2DM with macroalbuminuria group than that in T2DM with microlbuminuria ,T2DM with normalbuminuria and control groups.
our results revealed that circulating serum miR‐130b was inversely correlated to UACR(r = −0.759, p < 0.001),Scr(r = −0.759, p = 0.001)andBUN(r = −0.759, p < 0.001). ) and positive correlation with eGFR (r=0.373, p<0.001).
These results suggest that the decrease of serum miR‐130b may be closely associated with the pathological progress of DN . Bai et al 2016 reported that the miR-130b play critical role of in regulating renal tubulointerstitial fibrosis in diabetic nephropathy.
So,decreased serum miR-130b level can be considered as a good predictor for DN and indicate the existence of early renal damage in diabetic patients even before the appearance of albuminuria. in view of its reno-protective role by inhibiting lipid peroxidation reaction, chronic inflammation, renal fibrosis and extracellular matrix deposition .
As regard miR‐146a , Our study explored that the serum level of miR‐146a of was significantly increased in diabetic group more than that of the Pre-diabetic and control groups, With p value (< 0.001) .
Our results showed that serum miR‐146a was positively correlated with fasting blood glucose (FBG) levels (r = −0.627, p < 0.001), with HbA1c levels(r = −0.646, p < 0.001)and HOMA‐IR(r = −0.551 p < 0.001). In contary to Balasubramanyam et al 2011 that showed that miR-146a expression was negatively correlated to fasting blood glucose (P = 0.002), HbA1c (P = 0.002), insulin resistance (P = 0.001).
In addition, we found that serum miR‐146a exhibited different expression in T2DM patients with different stages of urinary albumin. The levels of circulating serum miR‐146a gradually and markedly increased and was significantly higher in T2DM with macroalbuminuria group than that in T2DM with microalbuminuria ,T2DM with normalbuminuria and control groups.
our results revealed that circulating serum miR‐146a was positively correlated to UACR(r = 0.724, p < 0.001),Scr(r = 0.436, p < 0.001)andBUN(r = 0.451, p <0.001),and was nagatively correlated to eGFR (r = −0.450, p < 0.001)
So , Change the levels of serum miR-130b( decreased) and miR-146a (increased) )in the early stage of DN, suggesting that they can be used as new diagnostic biomarkers for early detection of DN. Furthermore,further change during the progression of DN and correlated with eGFR and UACR, which are two clinical parameters that reflect the severity of renal impairment. These suggest the involvement of the two miRNAs in the pathogenesis and progression of DN. thus possibly making them use¬ful markers to monitor the progression of DN.