RU2437161C1 - Method for reservatrol-based osteoporosis correction and fragility fracture prevention - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: reservatrol efficacy is studied in female Wistar rats with simulated osteoporosis by bilateral ovariectomy. For osteoporosis correction, reservatrol is administered in animal intraperitoneally for eight weeks after ovariectomy daily once a day in single dose 2 mg/kg. Osteoprotective action of reservatrol is estimated by a bone tissue microcirculation level and width of trabeculas of bone.

EFFECT: improved blood circulation and microarchitectonics of bone tissue in hypoestrogenic 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 of preventing bone loss found by a number of authors when studying the osteoprotective and antitumor activity of resveratrol (Jen-Liang Su, Ching-Yao Yang, Ming Zhao, Min-Liang Kuo, Men-Luh Yen “Forkhead proteins are critical for bone morphogenetic protein-2 regulation and anti-tumor activity of resveratrol ", J Biol Chem. 2007 Jul 6; 282 (27): 19385-98).

The main disadvantages of the method is that there is no evaluation of the anti-osteoporotic effect of resveratrol in rats in vivo, the effect of resveratrol on the degree of blood supply to the bone tissue, as well as the width of the bone trabeculae, has not been 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.

The task is achieved by the fact 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, having antioxidant, estrogen , anti-aggregation and endotheliotropic activity, 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 was started 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 software version 3.8.1. 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.

An example of a specific implementation.

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 the development 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 with a hypoestrogenic state, leading to the occurrence of osteoporotic fractures.

In the group of animals that received resveratrol at a dose of 2 mg / kg intraperitoneally daily for eight weeks after ovariectomy, a significant increase in the average level of microcirculation in the proximal hip metaphysis was observed compared with the control group to 91.0 ± 12.8 PE. At the same time, there was a significant increase in the average width of the bone trabeculae compared with the animals of the control group up to 90.0 ± 2.0 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 (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 at a dose of 2 mg / kg for 8 weeks 91.0 ± 12.8 ** 90.0 ± 2.0 ** Note: * - p <0.05 in comparison with a group of intact animals; ** - p <0.05 compared with the group of control animals.

Thus, against the background of bilateral ovariectomy, resveratrol increases the level of microcirculation in the proximal hip metaphysis to the level of intact animals and, due to the 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 injected intraperitoneally daily with a dose of 2 mg / kg daily eight times after ovariectomy, evaluating osteoprotective the effect of resveratrol on the level of microcirculation in bone tissue and the width of bone trabeculae.