Study The Arginase Activity And Its Correlation With Liver And Kidney Functions For Patients With Type-2 Diabetes Mellitus in Nineveh

Contact: ali.ma@wecistanche.com

Ruaa Waleed Alramadhany, Hamodat, Zahraa Mohammed Al

Abstract

Background: Arginase plays an essential role in type 2 diabetes mellitus. Aim of the study: The study aimed to determine the activity of arginase in individuals patients with diabetes mellitus type 2 (DMT2) and study the correlation between arginase activity and the investigated variables Patients and material methods: The study included 201 adults divided into two groups: those with DMT2 (136 people) and control groups (65 people). Blood non-fasting persons had their determination of arginase activity, glucose, glycated hemoglobin (HbA1c), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, and creatinine levels. Results: The findings showed that the level of arginase activity in patients with DMT2 was significantly increased (p≤0.0001) in comparison with control groups, but there was no significant differentiation in the activity of arginase for both gender and age. A positive significant correlation (P=0.01) was found between Glucose (r=0.584), HbA1c (r=0.454), AST(GOT) (r=0.433), ALT (GPT) (r=0.295), ALP (r=0.639), Urea (r=0.667), and Creatinine (r=0.658). Conclusion: In patients with DMT2, increase the activity of arginase plays an active role through it is direct effect on liver and kidney functions.

Keywords: Arginase, diabetes mellitus type 2, Glucose, HbA1c %.

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1. Introduction

Diabetes mellitus is considered one of the common diseases that lead to death in the absence of followup of the disease, as it was found that diabetes is on the increase, and in a study that recorded in 2017, approx 462 million people worldwide were influenced by type 2 diabetes, accounting for 6.28 % of the global population (4.4 % of those aged 15-49 years, 15 % of those aged 50-69 years, and 22 %of those over the age of 70 years) [1].

Besides, diabetes mellitus has different types, the most important of which is type 1, called insulin-dependent Juvenile Diabetes. The second type, which is called adult diabetes, is not dependent on insulin [2]. Arginase (EC 3.5.3.1) catalyzes L-arginine hydrolysis into L-ornithine as well as urea and needs for its reaction cofactor, especially Mn+2. Arginase is a part of the urea cycle that regulates the L-arginine level [3]. Arginase is one of the important enzymes involved in the metabolism and formation of urea [4,5]. Arginase has two isozymes, the first is found in the cytosol in the liver, which plays a major role in the urea cycle, and the second is located in the mitochondria of the kidneys and prostate [4,6].

Besides, a study has found that patients with DMT2 have an increase in the activity of arginase [7], and in other studies conducted on rats exposed to diabetes, they have an increase in the activity of the arginase. It was also found that patients with DMT2 have a failure in liver function and a rise in activity of each AST, ALT, and ALP [8,9]. Furthermore, It was also found that diabetic patients get a rise in glycated hemoglobin, especially untreated patients, compared to treated patients and differences in liver functions. In addition, DMT2 suffers from impaired kidney function through high levels of urea and creatinine [10,11,12].

Our study aimed to measure the activity of arginase in the sera of patients with DMT2 and it is relationship to liver and kidney function.

2. Materials and Methods

2.1.population study:

The study was conducted on 201 patients who were divided into two groups, the first of which was 136 patients with type 2 diabetes of both sexes (78) females (58) males, and their ages ranged from (35-75). The healthy group included 65 people without Diabetes and other diseases of both sexes (33) females (32) males; their ages ranged from (35-68). Diabetic patients with other diseases such as heart, cancer, and kidney disease were excluded.

The study was conducted on diabetic patients who visit the clinics of Al-Wafa Center and the outpatient medical clinics licensed by the Nineveh Health Department. The patients were divided into two groups according to the age group, the first group was (35-45) years, and the second group was (≥46).

Furthermore, the Ethics Committee of the University of Mosul (College of Science) and the Nineveh Health Department followed the study protocol. All participants were provided with an informed consent questionnaire form to participate in the research.

2.2. Samples Collection:

2.2.1. Serum samples were obtained by drawing 5 ml of venous blood, then placing it in a gel tube and leaving it for 15 minutes at a temperature of 37Ċ and centrifuging 3000xg, then separating the serum and keeping it at a temperature of -20 Ċ until the necessary tests are performed.

2.2.2. Whole blood 2 ml of venous blood was withdrawn from only patients with diabetes mellitus type 2 to conduct a glycated hemoglobin test and placed in tubes containing an anticoagulant substance EDTA.

2.3. Variables Assay

2.3.1 Estimation of the arginase activityArginase activity was measured by following a modified method [13]. The activity of arginase represents the amount of enzyme needed to convert micromol from arginine to ornithine per minute.

2.3.2 Glycated hemoglobin measurements

glycated hemoglobin was measured using the German-made On-Call device, using the company's supplied kit G136-111.

2.3.3 Measurement of glucose concentration serum glucose concentration was measured using a colorimetric enzymatic method, using a kit manufactured by the Randox GL364 [13].

2.3.4 Measuring the activities of ALT, AST, and ALP. Using the Japanese-origin Fuji automated analyzer device and following the manufacturer's instructions

2.3.5 Measuring the concentration of urea and creatinine concentrations of urea were measured using the kit supplied by biosystems company [15] and creatinine was measured using the kit supplied by Biolabo Company [16].

2.4.Statistical Analysis

The analysis was done using SPSS version 25. The data were analyzed. Mean ± standard deviation (SD) was obtained; two groups were comparable with an independent T-test. The Pearson correlation coefficient is used to explore the correlation of serum arginase activity with the study's variables. The 0.0001 P-values were taken into account statistically significant[17].

3. Results and Discussion

Diabetes mellitus Type 2 (DMT2) is a common disease, and failure to monitor it and take the necessary treatment leads to death [18,19]. DMT2 leads to an imbalance in the body's functions, such as a destabilization in the functions of the liver and kidneys [20]. And in turn, it affects the rest of the body's functions as it affects the disorder of enzymes such as arginase [21,22,23]. Therefore, in this study, we decided to study the efficacy of the arginase activity and its correlation with the studied variables, included glucose, glycated hemoglobin (HbA1c), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, and creatinine.

Table 1 shows the characteristics of the population study; where the mean± SD age of the controls group and patients with DMT2 is 48.3±10.6 years and 50.7±10.3 years, respectively, and that the percentage of females (%57.4) is more affected by the disease than males (%42.6)activity is high and that insulin infusion can lower it [24]. Reduced substrate concentration is another important mechanism that could explain decreased nitric oxide (NO) generation. Arginase's accelerated enzymatic conversion of arginine to ornithine. Arginase is consistent with this nitrogen oxide species NOS vasodilatory properties are reduced. [24,26].

Some factors affecting arginase activity were also studied, including gender and age in patients and the control group. Table (2) and Figure 1 show that the sex factor does not affect arginase enzyme activity in patients and the control group. The arginase activity is higher in males than females in each of the patients and Control group, but there are no significant differences, there was no effect of the age factor on the activity of the arginase in patients and control group. However, the activity of arginase increases with age but without a significant difference.

Table (3) compares the studied variables, including glucose, glycated hemoglobin, liver and kidney functions in DMT2 patients and their comparison with controls. A significant increase compared with the control group. A significant increase (p≤0.0001) in the glucose concentration for patients compared to the control group agrees with ( Kashyap et al .2008) [24]. Also, increased HbA1c % agrees with (shatanawi et al. 2017)[24,26,27].

Studies have shown that there is a significant increase ( p≤0.0001) in AST, ALT, and ALP in patients with DMT2 compared to the control group, and this is consistent with [28,29]. This study is comparable to a study conducted by Salman et al [29] and the two additional studies conducted by Idris et [30], who reported an obvious rise in ALT and AST. Ni et al. Htet also conducted a study on patients with diabetes in Malaysia, and the study showed that these patients had liver function tests that were abnormal [31]. To supplement, Wang et al. had conducted a study in which they examined DMT2 patients and control. Their research discovered that ALT was significantly associated with a heightened risk of type 2 diabetes in the Chinese population [32]. This study discovered that ALT was significantly associated with an increased risk of DMT2 in Chinese individuals. The occurrence of these enzymes could be attributable to the significant hepatotoxic effect intense insulin resistance (measured as insulin resistance or HbA1c) may lead to higher levels of fatty acids (i.e., free fatty acids) in the liver [28,33]. This accumulation may be due to disruption of the cell membrane, malfunction of the mitochondria, and increased production of substances like transaminases, fatty acids, and toxic metabolites. These disruptions may also cause mitochondrial dysfunction, oxidative stress, and a rise in pro-inflammatory cytokines [34,29].

The results also showed in Table 3 the effect of the arginase activity on kidney function, which included urea and creatinine, where the results showed a significant increase ( p≤0.0001) for each of urea, creatinine in patients with DMT2 compared to the control group, this is consistent with other studies [35,36,37,38].

According to this study, high concentrations of urea and creatinine in the blood of diabetes patients indicate a pre-renal disease [36]. In 2017, Abdul Rahman Al-Dakhil conducted a study in Saudi Arabia that revealed a comparison [39]. Iraq had the highest prevalence of diabetic nephropathy among arab countries, whereas the United Arab Emirates had the lowest majority in Bahrain [39].

Correlation of arginase activity with variables study Table 4 and Figure 2 show the correlation of arginase activity was studied with the variables, which included glucose, HbAc%, AST, ALT, ALP, urea, and creatinine

1. Correlation of arginase activity with glucose results showed in Table (4) and Figure (2,1) that there is a positive relationship between the activity of arginase and the concentration of glucose in the blood. This result agrees with [40]. While our study does not agree with Shatanawi and Momani, 2017. Where they found that the levels of arginase do not show any significant relationship with the concentration of blood sugar [25].

2-correlation of arginase and HbA1c The results showed in Table (4) and Figure (2, 2) that there is a positive correlation between arginase activity and HbA1c. This is consistent with a previous study [25]. That found a positive correlation between the arginase activity and HbA1c because that chronic exposure to high glucose levels is necessary to increase arginase activity.

3-correlation of arginase and liver functions The results showed in Table (4) and Figure (2.3), (2.4) and (2.5) a significant increase and a positive correlation between the activity of arginase and liver function in patients with DMT2 compared to the control group AST, ALT, and ALP. This result is consistent with a study that found that the results of liver function tests for patients with DMT2 were significantly higher compared to the control group [41].

The positive correlation of arginase activity with these enzymes is due to the toxic effect of the liver due to the fatty acids that are formed in excess quantities on the liver cells as a result of insulin resistance [33].

3-correlation of arginase and kidney function urea and creatinine The results showed in Table (4) and Figure (2.6), (2.7) a significant positive correlation between the activity of arginase and kidney function in patients with DMT2. A study found that there was a agree with [42]. The findings of this study also revealed that poorly regulated blood sugar levels could result in high glucose levels and blood urea levels, putting diabetes individuals at a higher risk of developing nephropathy. This is in line with the findings of a prior study, which demonstrated that the most common cause of increasing kidney injury is hyperglycemia [36].

Conclusions

The increased activity of arginase in patients with DMT2 directly affects liver and kidney functions. Besides, disorders in liver and kidney function generally coincide with DMT2 diabetes in Nineveh Governorate. Therefore, screening for liver and kidney dysfunction in diabetic patients is required to prevent further complications associated with it with the liver and kidneys.

Acknowledgments

I extend my thanks and appreciation to my colleges' deanship (College of Science) and my department (Chemistry). As well as the Nineveh Health Department to facilitate the conduct of this research study.

Funding This study did not receive external funding.

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