MOROȘAN Elena1 , MITITELU Magdalena1*, UDEANU Denisa Ioana1, IONIṬǍ Ana Corina1, BLEBEA Nicoleta Mirela2, POP Anca Lucia1*,HOVANEṬ Marilena-Viorica3
1Clinical Laboratory and Food Hygiene Department, Faculty of Pharmacy, ”Carol Davila” University of Medicine and Pharmacy, 6, Traian Vuia Street, 020956, Bucharest (ROMANIA)
2Pharmacology and clinic Pharmacy Departament, Faculty of Pharmacy, Ovidius University from Constanta, Romania
3 Pharmaceutical Botany Department, Faculty of Pharmacy, ”Carol Davila” University of Medicine and Pharmacy, 6, Traian Vuia Street, 020956, Bucharest (ROMANIA)
*Corresponding author: magdamititelu@yahoo.com
Abstract
The research aimed to evaluate the serum concentrations of urea, creatinine, uric acid compared to the concentrations of the same parameters excreted in the urine during treatment. For a more detailed analysis considering the association of gout with other diseases such as diabetes, research also looked at the following blood parameters: blood glucose; cholesterol; serum triglycerides.
Keywords: gout, Polygoni Avicularis herba, Fraxini folium,
Introduction
Inflammatory diseases have affected the population since time immemorial. This determined the researchers to study them in detail and the treatments did not stop appearing. Their effectiveness has led to a decrease in the incidence of inflammatory diseases, however, lifestyle changes and technological developments have led the population to be exposed to a worryingly high number of risk factors for these diseases [1,2].
Gout is a metabolic disease that has affected the population since ancient times. Currently, it represents 1-2% of the causes of general mortality and 2-4% of rheumatic pathology. Gout is formed by the manifestation of a heterogeneous group of diseases that occur alone or in combination and includes: hyperuricemia, attacks of typical monoarticular inflammatory arthritis, deposits of urate crystals in and around the joints, interstitial deposits of urate crystals in the renal parenchyma, urolithiasis. Hyperuricemia, the main characteristic of gout, is defined by a plasma urate concentration greater than 7 mg / dL. This is the result of increased urate production, poor uric acid excretion or a combination of these two processes [3,4].
Clinical parameters are essential in this disease, both for establishing the diagnosis and for monitoring the evolution under treatment. They have broadened the area of knowledge of pathological processes and suggest the possibility of understanding the essence of these processes [5-9]. Gout is a common form of inflammatory osteoarthritis. Paradoxically or not, today we are facing an exacerbated increase in the incidence of gout similar to that of our ancestors hundreds of years ago. This is why we started the study of developing a treatment that is available to today’s population that adopts an unbalanced lifestyle [3,4,5].
We chose the vegetable products Polygoni Avicularis herba and Fraxini folium due to the diversity of active substances they contain. Plant extracts (depending on the solvent and extraction conditions used), due to the diversity of the composition, can influence some enzymatic systems in the sense of induction or inhibition and can influence the biochemical parameters [10]. The use of natural extracts in the treatment of various diseases must also take into account the quality of the environment where the raw materials come from. Numerous studies demonstrate the contamination of natural resources with pollutants in the development environment [11-16].
Materials and Methods
For the study were used batches of rats, male, Wistar breed, weighing 200 ± 10 g, purchased from the Bio.Mase U.M.F. “Carol Davila”, Bucharest. The animals brought from the farm were kept in the conditions of our laboratory for 2 days, in order to get used to the new habitat and the diet. The animals were fed twice a day and received water ad libitum.
The experiment was carried out in accordance with Directive 2010/63 / EU of the European Parliament and of the Council on the protection of animals used for scientific purposes and Ordinance no. 37 of the Government of Romania of 2/02/2002.
The diet consisted of feeding animals with food rich in animal protein (PRIMORDIAL Holistic Grain-Free Puppy with Chicken and Ocean Fish, nutritional analysis: moisture 8%, crude protein 30%, crude oils and fats 19%, crude ash 8%, raw fibers 2.6%.) And dark chocolate, according to the models proposed by the literature.
Lots were formed consisting of 10 rats that received 1 mL / 100g oral body daily from aqueous solutions, as follows:
- Lot I (control group) – Sodium chloride solution 0.9%;
- Lot II – Aqueous solution from Polygoni Aviculare grass 5%;
- Lot III – Aqueous solution of Fraxini folium 5%;
- Lot IV – Mixture of Polygoni Aviculare herba and Fraxini folium in mass ratio 1/1 concentration 5%;
The aqueous solutions were obtained by infusing 5 grams of crushed vegetable product in 200 ml of water for 20 minutes. The extracts were standardization in total polyphenols and total flavonoids. Two hours after the last administration, the animals were guillotined, blood was collected, and the serum separated by centrifugation was used to determine the biochemical parameters at the Cormay biochemical analyzer with specific reagent kits [17-20].
In order to determination the parameters monitored in the urine we followed the following steps:
Initially, the following parameters were measured. On the 4th day after the first administration of the solutions, two rats from each group were isolated in collection cages for 24 hours for diuresis. After collecting the necessary samples, the following parameters were analyzed: uric acid, urea, creatinine, urine volume, pH.
The same protocol was followed on days 7, 10 and 14 of the analysis.
On the 7th day of treatment, 3 rats from each group were placed in the diuresis equipment. Thus, we were able to track the relationship between the amount removed, time and the aqueous extract administered to all animals treated and to make an average of each group treated.
The variations of the effect at each moment compared to the initial moment were calculated according to the formula:
(Ex-E0)/E0 x 100,
where:
E0= the initial effect of the control;
Ex = the effect measured for each component studied
Results and Discussions
Results obtained from serum analysis
The determination of non-protein nitrogenous constituents was done by the Cormay method. For each parameter, 10 determinations were performed (Table 1,2,3).
Table 1. Influence of extractive solutions on serum urea in rats
Tested Product | Serum urea mg / dL | |
x±e.s. | Effect %/iniţial | |
Control group | 48,81 | – |
Group treated with Fraxini folium extract (Group F) | 44,61 | -8.60 |
Group treated with Polygoni Avicularis herba extract (Group P) | 44,19 | -9.46 |
Group treated with equimassal mixture extract (Group A) | 42,45 | -13.03 |
x±e.s.=media±standard error; p<0,05
Table 2. Influence of aqueous extractive solutions on uric acid in rats
Tested Product | Uric acid mg / dL | |
x±e.s. | Effect %/iniţial | |
Control group | 7.25 | – |
Group treated with Fraxini folium extract (Group F) | 6.19 | -14.62 |
Group treated with Polygoni Avicularis herba extract (Group P) | 6.23 | -14.06 |
Group treated with equimassal mixture extract (Group A) | 5.96 | -17.79 |
x±e.s.=media±standard error; p<0,05
Table 3. Influence of aqueous extractive solutions on serum creatinine in rats
Tested Product | Creatinine mg / dL | |
x±e.s. | Effect %/iniţial | |
Control group | 2.37 | – |
Group treated with Fraxini folium extract (Group F) | 2.19 | -6.16 |
Group treated with Polygoni Avicularis herba extract (Group P) | 2.22 | -6.32 |
Group treated with equimassal mixture extract (Group A) | 2.14 | -9.70 |
Serum test findings:
Following the treatment to which the groups of animals treated were subjected, the most significant effects compared to the initial time were recorded in the group treated with aqueous extract obtained from the equimassal mixture of the two plant products.
The results recorded on the analysis of non-protein nitrogenous serum constituents were -13.03% for serum urea, -17.79% for serum uric acid, -9.70% for serum creatinine.
The groups treated with aqueous extracts obtained from individual plant products of Polygoni Avicularis herba and Fraxini folium respectively decreased serum levels for urea, uric acid, creatinine which show the properties of plant products that contribute to lowering the factors of risk of gout and associated pathology
Results obtained from urine analysis
The determination of non-protein nitrogenous constituents was done by the Cormay method. For each parameter, 5 determinations were performed corresponding to treatment days 1, 3, 7, 10 and 14 (Figure 1, Figure 2, Figure 3).
Determination of the amount of urea in the urine:
Figure 1. Graphical representation of the influence of aqueous extractive solutions on urine from urine in rats
Determination of the amount of uric acid in the urine:
Figure 2. Graphical representation of the influence of extractive solutions on uric acid in rat
Determination of the amount of creatinine in the urine:
Figure 3. Graphical representation of the influence of aqueous extractive solutions on creatinine in urine in rats
Urine test findings:
Following the treatment to which the groups of animals treated were subjected, the most significant effects compared to the initial time were recorded in the group treated with aqueous extract obtained from the equimassal mixture of the two plant products.
The results recorded on the analysis of non-protein nitrogenous constituents in urine were + 10.71% for urea content, + 11.98% for uric acid content and + 10.09% for creatinine content after 14 days of treatment. The groups treated with aqueous extracts obtained from individual plant products of Polygoni Avicularis herba and Fraxini folium, respectively, recorded increases in the levels of urea, uric acid and creatinine in the urine during treatment, which shows the properties of plant products that contribute to reducing risk factors in the appearance of gout and associated pathology.
Conclusions
Gout is a disease that affects the population regardless of gender and the age at which it begins is lower as the patient adopts an unbalanced diet and a sedentary lifestyle.
In the current context of society, the risk factors to which we are exposed in daily life and the prevalence of inflammatory diseases, a pharmaceutical product based on the two herbal products in the form of infusion would ensure good adherence of patients to treatment.
The combination of the two plant products, Poligony Avicularis herba and Fraxini folium, in the same pharmaceutical preparation determines the mutual potentiation of the diuretic action with the favor of eliminating uric acid, creatinine and urea.
In all the animals studied, an improvement in health was observed with the course of treatment. This aspect gives hopes that by making a pharmaceutical product based on the two herbal products, results can be obtained in a wide range of patients with distinct characteristics.
The most significant results were obtained after the administration of the aqueous extract obtained from the equimassal mixture of the two vegetable products, Poligony Avicularis herba and Fraxini folium. Thus, following the study, it can be concluded that the development of a pharmaceutical product based on the eczema mixture of Poligony Avicularis herba and Fraxini folium will represent a progress in combating gout and inflammatory pathology. Along with these benefits, the product could also help in improving hyperglycemia, hypercholesterolemia and elevated serum triglyceride levels.
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