RU2462765C1 - Method for endothelial dysfunction correction by distant preconditioning in adma-like gestosis model - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves simulating gestosis in Wistar rats by the daily intraperitoneal introduction of L-nitro-arginine-methyl ester 25 mg/kg for 14th to 20th day of pregnancy. The simulated pathology is corrected by distant preconditioning (DP) from the 10^th to 20^th day of pregnancy by 10-minute femoral artery clamping at the level of an upper one-third of thigh followed by tissue reperfusion.

EFFECT: method provides the effective endothelial dysfunction correction in pregnant women suffering gestosis ensured by a manifested anti-ischemic effect of DP in the specific experimental environment and the specified DP regimen.

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Description

The invention relates to medicine, in particular to experimental pharmacology.

Closest to the claimed solution is a method of correcting endothelial dysfunction in pregnant animals using L-arginine, a substrate for the synthesis of nitric oxide, according to the description of experimental studies in the article: "Pharmacological correction of L-arginine" ADMA-eNOS-associated targets "in experimental preeclampsia" , Kuban Scientific Medical Bulletin, No. 1, 2010, pp. 85-92.

The main disadvantage of this method is that L-arginine is a direct competitor to ADMA and an increase in its concentration only leads to an increase in the substrate for the formation of NO and activation of NO synthase due to the competitive displacement of ADMA. One of the leading pathophysiological factors in reducing the activity of endothelial NO synthase (e-NOS) and the development of gestosis is placental ischemia. An increase in the activity of NO synthase can be achieved with anti-ischemic agents. Therefore, the results of the correction of endothelial dysfunction in an ADMA-like model of pathology in pregnant animals using L-arginine are unsatisfactory.

The objective of the invention is a method for the correction of endothelial dysfunction with an ADMA-like model of gestosis, including the reproduction of distant preconditioning with anti-ischemic action.

The objective is achieved by the fact that against the background of gestosis modeling in an experiment, an intraperitoneal injection of pregnant female Wistar rats for 7 days with an ADMA-like blocker of endothelial NO synthase - N-nitro-L-arginine-methyl ether (L-NAME) - at a dose of 25 mg / kg correction of endothelial dysfunction is carried out by reproducing distant preconditioning. This leads to a pronounced correction of endothelial dysfunction in the simulated pathology. This effect is associated with a pronounced anti-ischemic effect due to the isolation of a number of humoral factors: adenosine, bradykinin, apioids, etc., followed by activation of the mitochondrial and plasmole protein kinases. The final effector element is the mitochondrial and plasmalleal pool of ATP-dependent potassium channels. The realization of their protective potential is associated with membrane hyperpolarization due to their opening, activation of the nitric oxide synthesis system, and anti-apoptotic mechanisms. This leads to increased bioavailability of NO.

The method is as follows.

The experiments are carried out on white pregnant rats of Wistar female females weighing 250-300 g. N-nitro-L-arginine-methyl ether (L-NAME) is administered intraperitoneally at a dose of 25 mg / kg / day for 7 days (from 14 to 20 days pregnancy). Remote preconditioning was carried out from 10 to 20 days of pregnancy with a 10-minute clamping of the femoral artery with subsequent reperfusion. The compression of the femoral artery was performed by applying a cuff to the upper third of the thigh. The absence of a pulse on the lower leg arteries served as a control of the correctness of the cuff application.

On the 21st day of pregnancy under anesthesia (chloral hydrate 300 mg / kg), a catheter is inserted into the right carotid artery to record blood pressure (BP), bolus administration of pharmacological agents is carried out in the right femoral vein. Vascular tests for endothelium-dependent vasodilation (ESV) are performed - intravenous administration of acetylcholine (AH) at a dose of 40 μg / kg, and endothelium-independent vasodilation (ENZV) - intravenous administration of sodium nitroprusside (NP) at a dose of 30 μg / kg K ) - Patent No. 2301015 of June 20, 2007 (Bull. No. 17).

When statistical data processing is calculated, the average value, the standard deviation. Differences are considered significant at p <0.05.

Concrete example

The blockade of NO synthase caused by the 7-day administration of L-NAME to pregnant rats disrupted the relationship between vasodilating and vasoconstrictor mechanisms for regulating vascular tone, as evidenced by the results of vascular tests for endothelium-dependent relaxation (acetylcholine) and endothelium-independent (increase) sodium nitrossruss 1.28 ± 0.23 in intact pregnant animals up to 3.06 ± 0.32 * (p <0.05). In addition, a significant increase in systolic and diastolic blood pressure was observed from 125 ± 6.3 and 82.0 ± 5.8 to 183.1 ± 9.4 and 136.7 ± 7.4 mm Hg. respectively.

Daily reproduction of distant preconditioning from 10 to 20 days of pregnancy led to a significant decrease in QED to 1.60 ± 0.2, which is two times less than in the group of animals with the introduction of L-NAME (p <0.05), as well as correction of systolic and diastolic blood pressure - 142.1 ± 6.6 and 104.6 ± 7.1 mm Hg respectively.

Thus, the obtained results convincingly indicate a pronounced correction of endothelial dysfunction under conditions of modeling ADMA-like L-NAME-induced gestosis in pregnant rats by reproducing distant preconditioning.

Claims (1)

A method for correcting endothelial dysfunction in an ADMA-like gestosis model, comprising reproducing a gestosis model in Wistar rats daily, from 14 to 20 days of gestation, by intraperitoneal administration of L-nitro-arginine-methyl ester at a dose of 25 mg / kg, characterized in that for correction of simulated pathology daily, from 10 to 20 days of pregnancy, reproduce distant preconditioning by 10-minute clamping of the femoral artery at the level of the upper third of the thigh, followed by reperfusion.