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ECPB 2017, 78(2): 61–65
https://doi.org/10.25040/ecpb2017.02.061
Research articles

Determination of serum Fractions of Hydroxyproline at chronic toxic Hepatitis in Rats against the Background of Correction by Lisinopril and Glutargin

N. RIKALO, YU. BEREHOVENKO
Abstract

The results of an experimental study on the effect of lisinopril and glutargin on the dynamics of serum hydroxyproline fractions of immature rats with chronic tetrachloromethane hepatitis are presented in the article. Experimental studies were carried out on 60 non-linear white laboratory immature rats, with an initial body weight of 50-70 g. The animals were divided into 5 groups of 12 rats each in the beginning of the experiment. The first group is intact rats, the second group is with chronic toxic hepatitis modeled by the intragastric administration of an oily solution of CCl4 at a dose of 0.1 ml / 100 g of weight twice a week in combination with a 5 % solution of ethanol for eight weeks [Pat. 43704 Ukraine]. Animals of the third group in parallel with hepatotoxins daily for six weeks in the treatment and prophylaxis regimen were injected lizinopril in a dose of 20 mg / kg (Lizinopril LLC Astrafarm, Ukraine). Animals of the fourth group in parallel with hepatotoxins daily for six weeks in the treatment and prophylactic regimen, L-Glutargin-L-glutamate 0.75 g (Glutargin, Health, Ukraine) was intragastrally administered at 30 mg / kg. The animals of the fifth group received an intragastric 20 % CCl4 oil solution at a dose of 0.1 ml / 100 g of weight twice a week in combination with a 5 % solution of ethanol, as well as L-Glutargin-L-glutamate 0.75 g (Glutargin LLC, Health, Ukraine) at 30 mg / kg and lisinopril at a dose of 20 mg / kg (lisinopril «LLC Astrafarm», Ukraine). After the end of the experimental period, animals with euthanasia under the thiopental anesthesia were removed from the experiment by decapitation and blood samples were taken (for biochemical study). Blood was centrifuged for 15 min at a speed of 3000 rpm in a laboratory centrifuge OPn-3 with a plasma release for further biochemical studies. The content of free hydroxyproline, peptide-linked hydroxyproline and total hydroxyproline in serum was determined with these methods. The complex of biochemical studies was carried out on a semi-automatic biochemical analyzer «Vital Microlab-300» (USA) and an automatic biochemical analyzer «Beckman Coulter AU-480» (USA). Preparation of samples and determination of biochemical parameters were carried out in accordance with the instructions to the devices and reagents by the general methods. To evaluate the metabolic processes of the connective tissue, the content of free hydroxyproline and peptide-bound hydroxyproline was determined. The principle of the method is based on determining the optical density of the red chromogen, formed by condensation of oxidation products of hydroxyproline with para-dimethylaminobenzaldehyde. The increase in the content of precisely peptid-linked hydroxyproline, as the most important indicator of collagen formation, in the modeling of chronic toxic hepatitis, is natural, since this indicator is a biochemical marker of the intensity of fibrogenesis and a predictor of the unfavorable course of the disease. While the simultaneous administration of hepatotoxins with lisinopril significantly reduced the level of bound hydroxyproline, both at the expense of the free and peptide-linked fraction. With the combination of lisinopril and glutargin, the content of peptide-linked hydroxyproline was decreased. The results obtained, in our opinion, can be evaluated as a positive therapeutic effect of ACE inhibitors in fibrosis and liver cirrhosis. Given that, according to the literature, the activation of the renin-angiotensin-alsterone system is involved in the pathogenesis of fibrosis processes against the background of chronic liver pathology. A significant increase in the concentration of free and peptide-linked hydroxyproline in rats with chronic toxic hepatitis has been established, and it has been established that lisinopril as a monotherapy of experimental chronic toxic hepatitis, and also in combination with glutargin, significantly reduces the content of peptide-linked fraction of hydroxyproline as an important factor of fibrosis in the liver.

Keywords: serum hydroxyproline, chronic tetrachlortemtan hepatitis, rats

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