151 This work is licensed under a Creative Commons Attribution 4.0 International License. Correlation Study of Retinol Binding Protein4, Vitamin A with Liver Function Enzymes in Iraqi Fracture Patients with and without DM2 Abstract A fracture is a damage to bone tissue that causes damage to the tissue surrounding the bone and may penetrate the skin. Subjects and methods: the present study included (80) fractured Iraqi patients (and 40) patients with DM2 and (40) without DM2 and compared them with (40) healthy control. Patients and control are matched in age. This study showed a significant increase in retinol-binding protein 4 (RBP4) and a considerable decrease in Vit .A GPT and GOT in fracture patients with and without DM2. In addition, there was a significant negative correlation between RBP4 with (GPT and GOT) in fracture patients with DM2 and a significant positive correlation between RBP4 with (GPT and GOT) in fracture patients without DM2. Keywords: Fracture, DM2, RBP4, Vit.A, GPT, GOT, ALP. 1. Introduction Any bone in the body can be fractured [1]. A broken bone is a complete or partial break in the bone tissue's continuity. A significant percentage of bone fractures occur because of high force impact or stress. Type 2 diabetes is one of the most common metabolic diseases worldwide. It is caused by a combination of two essential factors: insufficient insulin synthesis by pancreatic cells and insulin-sensitive tissue failing to respond to insulin [2]. Vitamin A's significance in bone health has been thoroughly researched. Preformed retinol and provitamin A are the two types of vitamin A that may be eaten. Preformed retinol may be found in various animal-derived foods, including Doi: 10.30526/35.4.2830 Article history: Received 13 March 2022, Accepted 14 June 2022, Published in October 2022. Ibn Al-Haitham Journal for Pure and Applied Sciences Journal homepage: http://jih.uobaghdad.edu.iq/index.php/j/index Omar Yousif Majnun Department of Chemistry, College of Education for pure Science / Ibn-Al-Haitham, University of Baghdad,Iraq. Omar.Youssef1205a@ihcoedu.uobaghdad.edu.iq Anwar F. Altaie Department of Chemistry, College of Education for pure Science / Ibn-Al-Haitham, University of Baghdad, Iraq. anwar.f.m@ihcoedu.uobaghdad.edu.iq https://creativecommons.org/licenses/by/4.0/ mailto:Omar.Youssef1205a@ihcoedu.uobaghdad.edu.iq mailto:anwar.f.m@ihcoedu.uobaghdad.edu.iq IHJPAS. 53 (4)2022 152 dairy, liver, and eggs. Plant-based foods, such as fruits and vegetables, are high in provitamin A components such as alpha-carotene and beta-carotene. Provitamin A can be absorbed as an intact carotenoid or oxidized to the retinal and subsequently converted to retinol in the enterocyte. Retinol that has been performed, on the other hand, is absorbed directly from the skin [3]. RBP4 is a particular transporter of retinol from the liver to the periphery, and it comes from adipose tissue in smaller amounts. High RBP4 levels have been linked to inflammation and have been linked to metabolic syndrome, cardiovascular disease, type 2 diabetes, and insulin resistance in several investigations [4]. A liver profile includes the SGOT test, which is a blood test. Serum glutamic-oxaloacetic transaminase is one of two liver enzymes that it measures.Aspartate aminotransferase (AST) is the name given to this enzyme nowadays. The amount of liver enzyme in the blood is determined by an SGOT (or AST) test [5].The enzyme alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters in an alkaline environment, producing an organic radical and inorganic phosphate [6]. In healthy people, it predominantly comes from the liver, and bone iso-enzyme is a marker of bone formation that may be found (in osteoblasts) [7]. In Rheumatoid Arthritis (RA) patients, ALP may provide important information for determining fracture risk and monitoring osteoporosis [8]. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) values are often utilized as indicators of liver damage caused by fatty acid infiltration and inflammatory stimuli [9]. The most specific sign of this liver disease is alanine aminotransferase (ALT). According to recent research, increased ALT is linked to insulin resistance, type 2 diabetes, and metabolic syndrome [10]. Hepatic insulin resistance and eventual reduction in hepatic insulin sensitivity were linked to blood ALT concentrations [11]. AST and gamma glutamyltransferase concentrations, on the other hand, were unrelated to alterations in hepatic insulin action [12]. 2. Materials and methods Blood samples were collected from (80) fracture patients of ages in the range of (22-70) years Half of the number(40) suffered from type 2 diabetes mellitus, and the other half(40) were without diabetes, and (40) were healthy in the same age and sex with patients. All patients were enrolled at Al-Wasti teaching hospital in Baghdad during the study period from November to September 2021. Retinol binding protein 4 and vitamin A were estimated by ELASA kits (E-EL-H1581ELABSCIENCE, E-EL-0135ELABSCIENCE) in the United States and (GPT, GOT, ALP) by colorimetric method. 3. Statistical analysis The results of this investigation were presented as mean± SD. A t-test was employed to compare the patient and control groups. P values of 0.05 and 0.001 were also judged significant [13]. 4. Results and Discussion IHJPAS. 53 (4)2022 153 The recent study in table (1) and table (2) showed the concentration of (RBP4), Vit. A GPT AND GOT as mean ± SD in Iraqi patient fractures with DM2 (G2), without DM2 (G3), and healthy control (G1). The present study data showed a higher significant increase (P≥0.001) in G2 than in other groups. RBP4 (retinol binding protein 4) is a new adipokine that regulates insulin activity in various diseases [14]. The rise in (RBP4) might be attributed to an increase in insulin secretion to compensate for insulin resistance [15]. A metabolic disease involving retinol-binding protein (4) and type 2 diabetes is frequently associated with aberrant adipokine production [16]. RBP4 is an insulin signaling modulator; its levels are higher in those with insulin resistance or type 2 diabetes. It acts as a relay between inflammation and lipid metabolism in people transitioning from type 2 diabetes. As a result of the increased quantity of (RBP4) in fracture patients owing to inflammation, which is compounded by metabolic syndromes such as diabetes mellitus type 2 (DM2) and insulin resistance [17], table (2) also revealed a non-significant rise in (RBP4) in G2 compared to G3. Furthermore, in the circulatory system, (RBP4) is a lipocalin family member and the major transport protein of the hydrophobic molecule retinol, also known as vitamin A [18]. RBP4 activity is the most significant in the liver, which stores most of the body's vitamin A reserves as retinyl esters. Retinyl esters are hydrolyzed to retinol to mobilize vitamin A from the liver, which then binds to RBP4 in hepatocytes. So the increase in RBP4 concentration leads to a highly significant decrease in Vit. A, as shown in tables (1) and (2) in G2, then other groups. Vitamin A is a fat-soluble vitamin that humans require. It comprises retinol and its physiologically related metabolites [19]. It controls cell proliferation, differentiation, immune cell reproduction, and embryonic development [20]. A significant decrease in Vit. A might be the result of an increase in (RBP4). Tables (1) and (2) also showed the levels of liver enzymes (GPT, GOT, and ALP). There were significantly decreased concentrations in (GPT, GOT, and ALP) in G3 and G3, then G1. The enzymes alanine amino transfers (GPT) and aspartate amino transfers (GOT) are primarily located in the liver. Still, they can also be present in red blood cells, heart cells, muscle, and other organs such as the pancreas and kidney. The levels of GPT and GOT, are essential in diagnosing liver disorders. These enzymes are broadly dispersed in bodily tissues and are employed in diagnostics [21]. GPT, GOT, and ALP levels in the blood are proportional to the extent of tissue damage [22]. ALP in the blood can be used to determine whether or not a bodily tissue, particularly the heart and liver, has been harmed. ALP (alkaline phosphatase) is a membrane-bound enzyme that hydrolases monophosphate at a high pH and has been linked to metabolic bone disease [23]. So the levels of GPT, GOT, and ALP in serum can help people diagnose body tissues, especially the heart and liver. The levels of liver enzymes in all groups in a normal value lead to a healthy liver. IHJPAS. 53 (4)2022 154 Table 1. Concentration Study of RBP4,Vitamin A, GPT, GOT, ALP IN DM2, without DM2, Healthy Control P value Mean ± SD Parameters Without DM2 No. 40 With DM2 No. 40 Control No.40 0.001* 40.38 ±30.37 44.74 ± 30.17 9.75 ±13.46 RBP 4 (ng/ml) 0.001* 53.49 ± 5.36 39.07 ± 4.52 54.32 ± 3.78 VitaminA (Mg/dl) 0.005* 29.48 ± 5.05 30.44 ± 5.96 32.85 ± 2.27 GPT (IU/L) 0.039* 28.28 ± 6.18 30.54 ± 7.89 31.72 ± 2.50 GOT (IU/L) 0.033* 100.32 ±10.41 96.00 ±16.52 103.13±7.56 ALP (IU/L) *Significant Table 2. Compersion study of some biochemical parameters between patients groups. P value Mean ± SD Parameters Without DM2 No.40 With DM2 No.40 0.521* 40.38 ± 30.37 44.74 ±30.17 RBP.4 (ng/ml) 0.001* 53.49 ± 5.36 39.07 ± 4.52 VitaminA (Mg/dl) 0.438* 29.48 ± 5.05 30.44 ± 5.96 GPT (IU/L) 0.348* 28.28 ± 6.18 30.54 ± 7.89 GOT (IU/L) 0.167* 100.32 ±10.41 96.00 ±16.52 ALP (IU/L) IHJPAS. 53 (4)2022 155 Table 3.Correlation between RBP4 with Vit.A and liver enzymes in G2 Table 4. Correlation between PRB4 with Vit.A and liver enzymes in G3 ALP GOT GPT Vit.A RBP4 r = 0.327 r =-0.036 r =-0.312 r =0.006 P = 0.039* P = 0.02* P = 0.05* P = 0.68 ALP GOT GPT Vit.A RBP4 r = -0.712 r = 0.592 r = 0.563 r = 0.149 P = 0.005* P = 0.001* P = 0.001* P= 0.35 IHJPAS. 53 (4)2022 156 y = - 0.0618 x + 33.207 0 5 10 15 20 25 30 35 40 45 20 40 60 80 100 RBP4 DM2 Figure 1 .Correlation between RBP with DM2 and GPT with DM 2 with DM2 GPT with DM2 y = - 0.0746 x + 34.041 0 5 10 15 20 25 30 35 40 45 0 20 40 60 80 100 120 RBP4 DM2 Figure 2. Correlation between RBP4 with DM 2 and GOT with DM 2 GOT with DM2 GOT with DM2 IHJPAS. 53 (4)2022 157 y = 0.1791 x + 87.991 0 20 40 60 80 100 120 140 0 20 40 60 80 100 120 RBP4 DM2 Figure 3. Correlation between RBP4 with DM2 and ALP with DM2 y = 0.0935 x + 25.703 10 15 20 25 30 35 40 20 40 60 80 100 RBP4 without DM2 Figure 4.Correlation between RBP without DM2 and GPT without DM2 ALP with DM2 GPT whithout DM2 IHJPAS. 53 (4)2022 158 y = - 0.2438 x + 110.16 0 20 40 60 80 100 120 140 0 20 40 60 80 100 120 RBP4 without DM2 Figure 6. Correlation between RBP4 without DM2 and ALP without DM2 without DM2 ALP without DM2 y = 0.1204 x + 23.417 0 5 10 15 20 25 30 35 40 45 0 20 40 60 80 100 120 RBP without DM2 Figure 5. Correlation between RBP4 without DM2 and GOT without DM2 GOT whithout DM2 IHJPAS. 53 (4)2022 159 5. Conclusion This study concludes that retinol binding protein 4 has a good parameter for detecting the evolution of fracture. Reference 1. Maripuri SN, Rao P, Manoj-Thomas A, and Mohanty K. The classification systems for tibial plateau fractures: How reliable are they. Injur journal 2008, Y 39(10), 1216–1221. 2. Unai Galicia-Garcia , Asier Benito-Vicente , Shifa Jebari , Asier Larrea-Sebal ,Haziq Siddiqi, Kepa B. Uribe, Helena Ostolaza and César Martín. Pathophysiology of Type 2 Diabetes Mellitus. International journal moleculer science journal 2020, 21(17), 6275. 3. Michelle Min Fang Yee, Kok-Yong Chin, Soelaiman Ima-Nirwana and Sok Kuan Wong. Vitamin A and Bone Health: A Review on Current Evidence. International journal moleculer science 2021 , 26(6), 1757. 4. Bobbert P, Alice Weithäuser, Janin Andres, Thomas Bobbert, Uwe Kühl, Heinz Peter Schultheiss, Ursula Rauch and Carsten Skurk .Increased plasma retinol binding protein 4 levels in patients with inflammatory cardiomyopathy.Europ. Heart Failure Journal 2009, 11(12), 1163–1168. 5. Gjin Ndrepepa" Aspartate aminotransferase and cardiovascular disease—a narrative review. Journal of Laboratory and Precision Medicine .2021, 10.21037/jlpm-20-93. 6. Ganeb SS, El-Brashy AE, Baraka EA, Aboelazm AA, Basset SA. Peroxisome proliferator- activated receptor gamma expression in peripheral monocytes from rheumatoid arthritis patients. The Egyptian Rheumatologist journal .2016, 38(3):141-6. 7. Bover J, Ureña P, Aguilar A, Mazzaferro S, Benito S, López-Báez V, Ramos A, Cozzolino M. Alkaline Phosphatases in the Complex Chronic Kidney Disease-Mineral and Bone Disorders. Calcified tissue international. 2018, 14:1-4. 8. Sultan, N. S., & AL- Tai, A. F. Estimation of pentraxin-3(PTX3) in Rheumatoid arthritis males patients with (with and without) type II diabetes mellitus in Iraq. Ibn AL- Haitham Journal for Pure and Applied Sciences .2018, 31(3), 55–62. 9. Rector RS, Thyfault JP, Wei Y, Ibdah JA. Non-alcoholic fatty liver disease and the metabolic syndrome: an update. World Journal Gastroenterol .2008, 14,185–192. 10. Liangpunsakul S, Chalasani N. Unexplained elevations in alanine aminotransferase in individuals with the metabolic syndrome: results from the third National Health and Nutrition Survey (NHANES III). Am Journal Med Sci 2005, 329,111–116. 11. Oh SY, Cho YK, Kang MS, Yoo TW, Park JH, Kim HJ, Park DI, Sohn CI, Jeon WK, Kim BI, Son BH, Shin JH. The association between increased alanine aminotransferase activity and metabolic factors in nonalcoholic fatty liver disease. Metabolism journal .2006, 55,1604–1609. IHJPAS. 53 (4)2022 160 12. Sattar N, Scherbakova O, Ford I, O’Reilly DS, Stanley A, Forrest E, Macfarlane PW, Packard CJ, Cobbe SM, Shepherd J. The West of Scotland Coronary Prevention Study: Elevated alanine aminotransferase predicts new-onset type 2 diabetes independently of classical risk factors, metabolic syndrome, and C-reactive protein in the West of Scotland Coronary Prevention Study. Diabetes journal .2004, 53,2855–2860. 13. Alan, C. Elliott, Wayne, A. Woodward. Statistical Analysis Quick Reference Guidebook: With SPSS Examples, 1st Edition, 2007; ISBN-13: 978-1412925600 14. Al-taie, A.F., warkaa T. saloum ; Forat yahya mohsin shabil. Correlation study of retinol binding protein (4), nasfatin and thyroid hormoes in colorectal cancer Iraqi male patients. Plant archives journal 2020,2841_2846. 15. wuquan den, yuping, yanling zheng etal, serum retinol bindig protein (4) levels are resistance in newly diagnosed Chinese hyepertensive patients. Metabolic syndorom journal 2014, 10.1186/1758-5996-6-72. 16. Yang xiao, aiminxu, xiaoyan Hui and etal "circulating lipocalin-2 and retinol binding protein (4) are associated with intima media thickness and subelincal atnerosclerosis in patients with type 2 diabetes. Pone journal .2013, 0066607. 17. altaie, A.F.; warka T. saoum ; Eiman, A.A. Study on effect of methotrexate drug on retinol-binding protein 4, nesfatin and some trace elements in Iraqi male patients with rheumatoid arthritis. Biochem. Cell. Arch journal 2020, 0972-5075. 18. Julias, Steinhoff, Achimlass and Michael schupp "biological function of RBP4 and its relevance for human disease. Front. Physiol 2021, 10.3389. 19. Mc collum E.V.; Davis M. the necessity of certain lipids during growth. Journal Biol. chem .1973, 15, 167-175. 20. blaner, W.S. STRA6, a cell surface receptor for retinol binding protein, the plot thicknes. A Cell Press journa .2007, 5, 164- 166. 21. warkaa, T. ALs-saadi, A.F AL taie and forat yahya mohsin shabil , Correlation study of obestation and progranulin with liver function enzyme in Iraqi female patients with colorectal cancer. Planet archives 2020, 2412-2416. 22. xing – jiu Huang, yang-kyu choi and and hyung_soon im "aspartate aminotransfearase (AST/GOT) AND Alanine aminotransferase (ALT/GPT) detection techniques. Sensors journal .2006, 6, 756-782. 23. Masi, L.; marini, F. etal. Polymorphic variants of alkaline phosphatase gene correlate with clinical signs of adult hypo phosphatasia. Osteoporosis international journal .2021, 32, 2461-2472.