RU2437160C1 - Method for reservatrol and enalapril based osteoporosis correction and fragility fracture prevention - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: efficacy is studied in female Wistar rats. Osteoporosis is simulated by bilateral ovariectomy. Osteoporosis is corrected by intraperitoneal introduction of reservatrol 2 mg/kg and intragastric introduction of enalapril 0.5 mg/kg daily once a day for eight weeks after ovariectomy. Osteoprotective action of reservatrol combined with enalapril is estimated by a bone tissue microcirculation level and width of trabeculas of bone.

EFFECT: improved blood circulation and microarchitectonics of bone tissue in hypoestrogen condition.

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Description

The invention relates to medicine, in particular to experimental pharmacology, and can be used to correct osteoporosis and prevent the occurrence of osteoporotic fractures.

Closest to the claimed solution is a method for correcting osteoporosis and preventing the occurrence of osteoporotic fractures with enalapril (Pat. RU 2369390, IPC ⁷ A61K 31/401. A method for correcting osteoporosis and preventing the occurrence of osteoporotic fractures with enalapril / Pokrovsky MV, Gudyrev OS, Veitelson A.V. et al .; applicants and patent holders Pokrovsky M.V., Gudyrev O.S., Faitelson A.V. - No. 2008122134/14; filed 02.06.08; publ. 10.10.09. Bull. No. 28).

The main disadvantages of the method is that there is no evaluation of the anti-osteoporotic effect of the combination of enalapril and resveratrol in female Wistar rats, the effect of this combination of drugs on the degree of blood supply to bone tissue, as well as the width of the bone trabeculae, was not studied.

The objective of the invention is to develop a method for the correction of osteoporosis and prevention of osteoporotic fractures, including the use of resveratrol in combination with enalapril.

This object is achieved in that, against the background of modeling osteoporosis in an experiment using bilateral ovariectomy in female Wistar rats, it is corrected for eight weeks by daily, once a day, intraperitoneal administration of resveratrol at a dose of 2 mg / kg, which has antioxidant, estrogenic, antiaggregatory and endotheliotropic activity, and intragastric administration of enalapril at a dose of 0.5 mg / kg, which, affecting the activity of the renin-angiotensin-aldosterone system (RAAS), inhibits the conversion of angiot of benzin I to angiotensin II, which in aggregate directly reduces the vasoconstrictor activity of circulating and tissue components of RAAS and leads to peripheral vasodilation, causes an increase in blood supply to bone tissue and an increase in the width of bone trabeculae,

The method is as follows.

The experiments were carried out on female Wistar rats weighing 200-250 g. An ovariectomy was performed under anesthesia (a solution of chloral hydrate intraperitoneally at a dose of 300 mg / kg at the rate of 0.1 ml of solution per 100 g of animal weight). Intraperitoneal administration of resveratrol at a dose of 2 mg / kg and intragastric administration of enalapril at a dose of 0.5 mg / kg began on the day after surgery and was carried out daily for eight weeks. On day 57 from the start of the experiment under anesthesia (chloral hydrate intraperitoneally at a dose of 300 mg / kg), microcirculation was measured in the proximal metaphysis of the right thigh. The microcirculation level was measured using Biopac systems equipment: MP100 polygraph with LDF100C laser Doppler flowmetry module and TSD144 invasive sensor, and AcqKnowledge version 3.8.1 software. After measuring microcirculation, the femurs were taken for histological examination; histological preparations of proximal femur metaphyses were stained with hematoxylin and eosin. Optical microscopy of the preparations was carried out using a Leica CME microscope (magnification × 100) with photographing of the preparations using an Olympus SP-350 camera (resolution 3264 × 2448). A morphometric study of the obtained photographs was performed using the ImageJ program version 1.39, during which the width of the bone trabeculae was estimated.

The obtained values were compared with intact animals (false ovariectomy operation), as well as control animals, in which generalized osteoporosis was simulated by bilateral oophorectomy.

When statistical data processing was calculated, the average value, the standard error. Differences were considered significant at p <0.05.

EXAMPLE OF SPECIFIC PERFORMANCE

The average level of microcirculation in the proximal hip metaphysis in intact (false-operated) animals was 100.5 ± 4.4 perfusion units (PE); the average width of the bone trabeculae was 97.7 ± 1.0 μm (table 1).

Bilateral oophorectomy eight weeks after the start of the experiment in control animals led to a significant decrease in the average level of microcirculation compared with intact ones to a value of 61.5 ± 3.7 PE. There were signs of osteoporosis: the average width of the bone trabeculae was 61.7 ± 1.2 μm (table 1). The discovered fact of a decrease in blood supply in bone tissue is an additional factor in the development of osteoporosis in the hypoestrogenic state, which leads to the occurrence of osteoporotic fractures.

In the group of animals, which received intraperitoneal resveratrol at a dose of 2 mg / kg and intragastrically enalapril at a dose of 0.5 mg / kg intravenously for eight weeks after ovariectomy, a significant increase in the average level of microcirculation in the proximal hip metaphysis was observed compared to the control group to 108. 6 ± 19.0 PE At the same time, there was a significant increase in the average width of the bone trabeculae compared to the animals of the control group to 94.2 ± 2.9 μm (table 1).

Table 1. Dynamics of indicators of the average level of microcirculation in the proximal metaphysis of the thigh and the average width of the bone trabecula in modeling osteoporosis and its correction with resveratrol in combination with enalapril (M ± m). Groups of animals The average level of microcirculation, PE The average width of the bone trabecula, microns Intact (false-operated) 100.5 ± 4.4 97.7 ± 1.0 Untreated (control) 61.5 ± 3.7 * 61.7 ± 1.2 * Receiving resveratrol 2 mg / kg and enalapril 0.5 mg / kg for 8 weeks 108.6 ± 19.0 ** 94.2 ± 2.9 ** Note: * - p <0.05 in comparison with a group of intact animals; ** - p <0.05 compared with the group of control animals.

Thus, resveratrol in combination with enalapril against the background of bilateral ovariectomy increases the level of microcirculation in the proximal metaphysis of the thigh to the level of intact animals and, due to increased blood supply to bone tissue, slows down the development of osteoporosis in the hypoestrogenic state, which, in turn, reduces the likelihood of osteoporotic fractures.

Claims (1)

A method for correcting osteoporosis and preventing the occurrence of osteoporotic fractures, characterized in that, against the background of modeling osteoporosis in an experiment using bilateral ovariectomy, female Wistar rats are administered intraperitoneally once daily daily at a dose of 2 mg / kg and intragastrically enal for eight weeks after ovariectomy. dose of 0.5 mg / kg, evaluating the osteoprotective effect of resveratrol in combination with enalapril by the level of microcirculation in bone tissue and the width of the bone trabeculae.