Method for reservatrol-based osteoporosis correction and fragility fracture prevention

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.

1 tbl, 1 ex

 

The invention relates to medicine, in particular to experimental pharmacology, and can be used for the correction of osteoporosis and the prevention of osteoporotic fractures.

Closest to the claimed solution is the way to prevent bone loss detected by a number of authors in the study osteoprotective and antitumor activity of resveratrol (Jen-Liang Su, Ching-Yao Yang, Ming Zhao, Min-Liang Kuo, Men-flood 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 this method is that no assessment antiosteoporotic actions of resveratrol in rats in vivo, has not been conducted to study the effect of resveratrol on the degree of blood supply to the bone tissue, as well as the width of bone trabeculae.

The objective of the invention is to develop a method for correcting osteoporosis and the prevention of osteoporotic fractures involving the use of resveratrol.

This object is achieved in that the background modeling of osteoporosis in the experiment with bilateral oophorectomy in female rats Wistar held its correction within eight weeks of daily, once a day, intraperitoneal introduction of resveratrol at a dose of 2 mg/kg, which, having antioxidant, estrogenic, antigrav the operating and endotheliotropic activity causes an increase in blood supply to the bone tissue and increases the width of bone trabeculae.

The method is as follows.

Experiments were performed on rats female Wistar weighing 200-250 g Oophorectomy was performed under General anesthesia (solution of chloral hydrate intraperitoneally at a dose of 300 mg/kg at a rate of 0.1 ml per 100 g weight of the animal). Intraperitoneal administration of resveratrol at a dose of 2 mg/kg was started the next day after surgery and was performed daily for eight weeks. 57 day from the beginning of the experiment under anesthesia (chloral hydrate intraperitoneally at a dose of 300 mg/kg) was performed measurement of microcirculation in the proximal metaphase right thigh. The level of microcirculation was measured using the company's equipment Biopac systems: the polygraph MR module laser Doppler flowmetry LDF100C and invasive sensor TSD144, and program AcqKnowledge version 3.8.1. After measuring microcirculation was made fence thighs for histological examination; histological specimens of the proximal metafit thighs were stained with hematoxylin and eosin. Was carried out by optical microscopy preparations with a microscope Leica CME (magnification ×100) with photographing preparations using camera Olympus SP-350 (resolution of 3264×2448). Morphometric study of the obtained images was made at the same time the program ImageJ version 1.39, in the course of which was estimated width of bone trabeculae.

The obtained values were compared with intact animals (false operation oophorectomy), as well as the control animals, which through bilateral oophorectomy simulated generalized osteoporosis.

When the statistical processing of the data was calculated the average value, the magnitude of the standard errors. The differences were considered significant at p<0,05.

An example of a specific implementation.

The average level of the microcirculation in the proximal metaphase thigh intact (false-operated) animals amounted to 100.5±4,4 perfusion units (PU); the average width of bone trabeculae was equal 97,7±1,0 µm (table 1).

Bilateral oophorectomy eight weeks after the start of the experiment led control animals to a significant decrease in the average level of the microcirculation compared with intact up to the value of 61.5±3,7 PE. Attended the signs of osteoporosis: the average width of bone trabeculae was 61,7±1.2 µm (table 1). Found the fact that reduction of blood supply to the bone is an additional factor in the development of osteoporosis at hypoestrogenism condition that leads to the emergence of osteoporotic fractures.

In the group of animals receiving daily for eight weeks after oophorectomy NR is drybrushing resveratrol at a dose of 2 mg/kg, there was a significant increase in the mean level of the microcirculation in the proximal metaphase hip 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 bone trabeculae compared with control animals to 90.0±2,0 µm (table 1).

Table 1
Dynamics of the average level of the microcirculation in the proximal metaphase hips and an average width of bone trabeculae in the simulation of osteoporosis and its correction by resveratrol (M±m).
Groups of animalsThe average level of the microcirculation, NEThe average width of bone trabeculae, mcm
Intact (false-operated)100,5±4,497,7±1,0
Not treated (control)61,5±3,7*61,7±1,2*
Received within 8 weeks of resveratrol at a dose of 2 mg/kg91,0±12,8**90,0±2,0**
Note: * - p<0,05 in comparison with the group is Oh intact animals;
** p<0,05 in comparison with the group of control animals.

Thus, resveratrol on the background of bilateral oophorectomy increases the level of microcirculation in the proximal metaphase thigh almost to the level of intact animals and by enhancing the blood supply to the bone tissue, slows down the development of osteoporosis at hypoestrogenism state, which, in turn, leads to a decrease of the probability of occurrence of osteoporotic fractures.

Treatment of osteoporosis and prevention of occurrence of osteoporotic fractures, characterized in that the background modeling of osteoporosis in the experiment with bilateral oophorectomy, females of Wistar rats for eight weeks after oophorectomy administered daily once a day intraperitoneally resveratrol at a dose of 2 mg/kg, evaluating osteoprotective effect of resveratrol on the level of microcirculation in bone and width of bone trabeculae.



 

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17 cl, 2 tbl

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1 tbl, 2 ex

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