European Journal of Cardiovascular Medicine

Sickle cell disease (SCD) is very commonly found in African populations. It is measured  that, 1 in 365 African American infants have sickle cell disease while 1 in 13 has are born with the sickle cell trait. The 2010 nationwide Centre for Disease Control (CDC) survey of state newborn screening programs which screen for sickle cell trait (SCT) reported that incidence of SCT was73.1 cases per 1000 black infants screened, 3.0 cases per 1000 white infants screened and 2.2 cases per 1000 Asians, native Hawaiian or other Pacific Islander infants have also screened. ^[1]   

In India, it was first discovered by Lehmann and Cut bush about (1952) 64 years ago. It had been discovered among the tribals of Nilgiri hills in southern India, but it commonly found in Chhattisgarh, tribes of Madhya Pradesh, Rajasthan and Maharashtra in central part of India. ^[2,3] 

Sickle Cell Disease (SCD) refers to a collection of inherited blood related disorders which was characterized by abnormal hemoglobin (Hb) molecules (National Institutes of Health, 2002). Haemoglobin is the conjugated proteineous compound, (heme is a prosthetic group and globin as a protein part) within erythrocytes (i.e. RBCs), which is involved for collection of O₂ (oxygen) in the lungs and transporting it throughout the body. The signs and symptoms of sickle cell disease are caused by the sickling of red blood cells.

The painful episodes can occur when sickled red blood cells (stiff, inflexible) are get stucked in the small blood vessels. These episodes decreases organs and tissues of oxygen-rich blood and can causes to organ damage, especially in the spleen, kidneys, lungs, and brain.  ^[4] 

The routine hematological and biochemical parameters; Hb, Hct, Iron, TIBC, MCV, MCHC, RET (Reticulocyte), RED, WBC, PLT (Platelet counts) were showed the variation in the sickle cell disease. LFT and KFT were also varies from the control. ^[5]

This study was being conducted in the Department of Biochemistry, People’s College of Medical Science and Research Centre (PCMS & RC) and Centre for Scientific Research and Development (CSRD), People’s University, Bhanpur, Bhopal. The study protocol had been approved by Departmental Doctoral Committee, Research Advisory Committee, Institutional Ethical Committee (Ref: PCMS/OD/2016/2551) and University Doctoral Committee of our institute/university.

Study Design:

The study was hospital based case control study, the sample size had been calculated by the expert statistician.

All the selected and enrolled patients had been confirmed with their stability state, without blood transfusion of four months prior to blood drawn. In addition, the patients were included in the study didn’t show infections, hospitalization or vaso-occlusive event, and were not under antibiotics therapy, corticosteroids or hyroxyurea (HU) treatment. 

The test group consists of 111 patients and control group consisted of 111 healthy individuals. All procedures followed were being in accordance and approved by the Research Ethics Committee of the People’s College of Medical Sciences & Research Center, Bhopal, and also with the Helsinki Declaration of 1975 and its revisions. Informed consent forms will be obtained from all patients.

Inclusion criteria for normal healthy subjects:

 Normal healthy person were comprised of departmental staff, medical students and the relatives who were healthy and accompany their OPD or IPD wards. And their well health condition was being detected.

Inclusion criteria:

1. A) Patients more than 5years age and less than 80 years of age.
2. B) Normal healthy individuals were recruited for the control group

Exclusion criteria:

1. A) Patients less than 5years and more than 80 years.
2. B) Individuals with any other hemoglobinopathy, and the patients having any history of blood transfusion within 3 months.
3. C) SCD patients under chemotherapy treatment were excluded from the study.

Screening tests for the sickle cell disease subjects:

The following screening tests had been carried out with SCD patients for the confirmation of sickle cell RBCs.

1. Sickling test with 2% meta-bisulphite: It is the principle of sickling test, was based on microscopical observation of sickling of red blood cells, when exposed to a low oxygen tension.
2. Solubility test with 0.02% sodium dithionate: It is the principle of solubility method, based on turbidity created when Hb S is mixed with sodium dithionate.
3. Peripheral blood film method: Thin blood films, stained with giemsa stain were examined by light microscopy (×100).
4.  Hb electrophoresis: The cellulose acetate membrane Hb electrophoresis method had been used to determine the presence of Hb-S in the sample. 
5. Patient’s history and blood cell counts such as; RBC, WBC, and HCT, MCH, MCV, MCHC, was being carried out.

To confirm any diagnosis, a sample of blood was examined under a microscope to check for large number of sickle cells, patient’s history and blood cell counts i.e.  RBC, WBC, and HCT, MCH, MCV, MCHC, was also carried out. Blood urea was estimated by Diacetyl Monoxime Method ^{[6, 7, 8 ]} and serum creatinine ^{[9, 10]} was estimated by Jaffe’s method.

Statistical Analysis

For the statistical analysis, statistical package SPSS 24 and Microsoft Excel 2010 had been used in this study.  The independent student ‘t’ test had been also applied for the comparison purpose, and “p” value ˂0.05 had been used for the level of significance.

Only few selective variables of kidney function tests were performed in the present study, which showed in table No-1. The selected variables blood urea and serum creatinine levels were showing variations:

The blood urea level was increased significantly high in cases of Sickle cell disease (41.02 ± 5.38; 29.27 ± 3.86) compared to controls and the serum creatinine level also elevated (1.39 ± 0.29; 0.77 ± 0.17) significantly high in cases compared to controls.

Table 1: Kidney Function Tests in Sickle Cell Disease

In graphical presentation both the variables were found highly significant in the cases compared to controls in SCD.

Graph – 1: Blood Urea level variation in Sickle Cell Disease

Graph 2:  Serum Creatinine level variation in Sickle Cell Disease

Silva Junior et al., (2013) studied that the, Kidney in sickle cell disease includes different types of glomerular and tubular disorders, which are associated with increased mortality. ^[11] Okafor UH et al. (2013) stated that the pathophysiology of sickle cell nephropathy (SCN) is related to the low oxygen tension, low pH, and high osmolality, these conditions in SCD patients predispose to red blood cell sickling. ^[12]   Marouf R et al., (2006) stated that sickle cell disease is associated with many functional and structural abnormalities of the kidney which may progress to end-stage renal disease. ^[13] 

In our study, there was highly significant difference in both serum concentration of urea and creatinine between patients and controls. This is similar to the findings of Silva Junior GB (2017) et al. ^[11]  and Pandey et al. ^[14]  but it is in contrast to that of al-Naama et al., ^[15] 

Blood urea in SCD cases and controls were found (41.02 ± 5.38; 29.27 ± 3.86) and Serum Creatinine in SCD cases and controls were found (1.39 ± 0.29; 0.77 ± 0.17).

Al-Naama LM et al., Christopher BE ., Edgar VL and Aloni MN., et al. reported that the, High serum urea and serum creatinine levels were found more during crises (23%. urea, 43%. creatinine) as compared to steady state (urea 3.3%, creatinine 15%) indicating reversibility of involvement after crisis. ^{[15, 16, 17, 18]} 

Aleem A. et. Al., (2008) studied that, the serum creatinine levels were found high in 73 patients with SCD at King Khalid University Hospital, Riyadh, Saudi Arabia. Creatinine clearance was found low in 12 patients (22.5%) and 7 of these patients had low and low-normal serum creatinine despite reduced creatinine clearance. Decreased serum creatinine level was found common in 28 patients (38%). ^[19]  Lakkakula BV et al (2017) studied bold urea and serum creatinine levels in sickle cell disease patients and they found that, sreum creatinine level were low, blood urea level were low, and but there was the increased uric acid level are found more commonly in SCD children than the controls. ^[20] 

Silva Junior GB et al studied that the serum creatinine levels in male and female SCD patients were found significantly lower. ^[21].

Voskaridou E, Terpos E, Michail S.  In their study they observed that, there were statistically significant positive correlations between blood urea serum creatinine levels, β-2 microglobulin and there were statistically significant negative correlations between β-2 microglobulin and Hb%. ^[22]

In kidney function tests, the Blood Urea level and Serum Creatinine levels had been found significantly increased in the sickle cell disease.  So, these KFT parameters could be helpful for the diagnosis of sickle cell disease.

1. Ojodu J, Hulihan MM, Pope SN, Grant AM; Centers for disease Control and Preventation (CDC), Incidence of sickle cell trait – United States, 2010. MMWR Morb Mortal Wkly Rep, 2014; 63:1155 – 1158. [PMID:25503918]
2. Lehmann H. and Cutbush M., Sickle-cell Trait in Southern India “Br Med J. 1952; 1(4755):404–405.doi: 10.1136/bmj.1.4755.404PMID: 14896162.
3. Vikas K. Gupta, Nilima Singh, S.W. Masram, S. K. B. Patil; Status of some Hematological and Antioxidant Parameters in SCA patients of Chhattisgarh Region, Journal of Medical Science and Clinical Research, 7, July 2015. Vol.03.

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https://www.ncbi.nlm.nih.gov/pubmed/15683091

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https://www.ncbi.nlm.nih.gov/pubmed/19151898; doi: 10.1100/tsw.2009.10.

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