RU2384894C1 - Method for modelling osteoporosis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to experimental medicine and concerns modelling osteoporosis. That is ensured by oophorectomy in a laboratory animal. Then omeprazole is administered in a dosage 0.3-0.4 mg/kg orally within 6 months.

EFFECT: higher accuracy of modelling osteoporosis caused by inhibitors of proton pump.

3 ex

Description

The invention relates to medicine and can be used to model osteoporosis.

There is a known method for modeling avascular osteonecrosis, taken as an analogue (Zaitseva M.Yu., Netylko GI Experimental modeling of avascular osteonecrosis of the condyles of the femur and tibia. Traumatol. Orthopedist. Russia. 2005; 37 (3): 36-39).

A known method of modeling osteoporosis by introducing steroids, adopted as a prototype (Wang G., Cui Q., Balian G. The patogenesis and prevention of steroid induced osteonecrosis Clin. Orthop. 2000; 370: 295-310).

However, the known method does not allow to simulate osteoporosis that occurs when exposed to proton pump inhibitors in different age groups.

The aim of the invention is to improve the accuracy of modeling of osteoporosis that occurs when exposed to proton pump inhibitors.

The technical result is achieved by the fact that the animals undergo bilateral oophorectomy followed by the introduction of omeprazole at a dose of 0.3-0.4 mg / kg orally for 6 months.

The method is implemented as follows.

Female Wistar rats weighing 220-250 g, which were kept on a standard diet and free access to food and water under vivarium conditions, underwent bilateral ovariectomy and omeprazole at a dose of 0.3-0.4 mg / was administered orally for 6 months / kg

Phosphorus, calcium, creatinine, albumin, alkaline phosphatase were determined in blood serum; in the bone tissue, the level of calcium and phosphorus was determined. Morphological examination of bone tissue was performed with preliminary decalcification. Tissue samples were fixed according to standard methods and stained with hematoxylin and eosin.

The method is confirmed by the following examples.

Example 1

The Wistar rat weighing 220 g (female) was kept on a standard diet and free access to food and water in vivarium conditions. Bilateral oophorectomy was performed as follows: in the conditions of the surgical stage of nembutal anesthesia (40 mg / kg), a median laparotomy was performed, uterine tubes were isolated by taking them on a ligature, and ovarian arteries were ligated. The ovaries were isolated and crossed the proximal end of the tubes behind the ligature. The resected end of the ligature tube was immersed in the abdominal cavity. A thorough hemostasis was performed. The aponeurosis and skin were sutured with a double suture. Omeprazole at a dose of 0.3 mg / kg was orally administered for 6 months.

Six months later, under the conditions of the surgical stage of nembutal anesthesia (40 mg / kg), blood was taken (1.5 ml), centrifuged (3000 rpm). The serum was then separated into cuvettes for determination of phosphorus, calcium, creatinine, albumin, alkaline phosphatase on an Olimpus analyzer. Next, bone tissue was collected in the area of the diaphysis of the femur and sternum and the level of calcium and phosphorus was determined. Morphological examination of bone tissue was performed with preliminary decalcification. Tissue samples were fixed according to standard methods and stained with hematoxylin and eosin.

The results of studies of biochemical parameters in an animal with ovariectomy and oral administration of omeprazole showed that the serum calcium level was 2.57 mmol / L (norm 2.16), alkaline phosphatase - 168.0 (norm 45.5), phosphorus - 1 82 (norm 1.6), albumin - 33.0% (norm 35), creatinine - 45.0 (norm 72). That is, there was an increase in the level of calcium, phosphorus and alkaline phosphatase, which indicates the development of osteoporosis.

The calcium content in bone tissue was 0.99 mg / g of tissue (normal 3.5 mg / g), phosphorus - 0.91 mg / g of tissue (normal 1.5 mg / g), which indicates a decrease in saturation of calcium and phosphorus bone tissue.

A morphological study revealed thinning of bone beams and degeneration of osteoblasts.

Example 2

A 250 g Wistar rat (female) was kept on a standard diet and free access to food and water under vivarium conditions. Bilateral oophorectomy was performed under the conditions of the surgical stage of nembutal anesthesia (40 mg / kg). A median laparotomy was performed, uterine tubes were isolated, taking them on a ligature, ovarian arteries were ligated. The ovaries were isolated and crossed the proximal end of the tubes behind the ligature. The resected end of the ligature tube was immersed in the abdominal cavity. A thorough hemostasis was performed. The aponeurosis and skin were sutured with a double suture. Omeprazole at a dose of 0.4 mg / kg was orally administered for 6 months.

Six months later, under the conditions of the surgical stage of nembutal anesthesia (40 mg / kg), blood was taken (1.5 ml), centrifuged (3000 rpm). The serum was then separated into cuvettes for determination of calcium, phosphorus, alkaline phosphatase, creatinine, albumin on an Olimpus analyzer. Next, bone tissue was collected in the area of the diaphysis of the femur and sternum and the level of calcium and phosphorus was determined. A morphological study of bone tissue was performed with preliminary decalcification in a 10% solution of nitric acid for 1 day. Tissue samples were fixed according to standard methods and stained with hematoxylin and eosin.

Biochemical parameters in an animal with ovariectomy and oral administration of omeprazole were as follows: serum calcium level was 2.75 mmol / L (norm 2.16), alkaline phosphatase - 204.0 (norm 45.5), phosphorus - 2.10 (norm 1.6), albumin - 30% (norm 35), creatinine - 58.0 (norm 72). That is, there was an increase in the level of calcium, phosphorus and alkaline phosphatase, which indicates the development of osteoporosis.

The calcium content in bone tissue was 0.91 mg / g of tissue (normal 3.5 mg / g), phosphorus - 0.71 mg / g of tissue (normal 1.5 mg / g), indicating a decrease in saturation of calcium and phosphorus bone tissue.

A morphological study found that the severity of thinning of bone beams and degeneration of osteoblasts is higher than in example 1.

Example 3

A Wistar rat weighing 240 g (female) was kept on a standard diet and free access to food and water under vivarium conditions. Bilateral oophorectomy was performed as follows: under the conditions of the surgical stage of nembutal anesthesia (40 mg / kg). A median laparotomy was performed, uterine tubes were isolated, bandaging them in the proximal (ovarian) section, and ovarian arteries were ligated. The ovaries were isolated and crossed the proximal end of the tubes behind the ligature. The resected end of the ligature tube was immersed in the abdominal cavity. A thorough hemostasis was performed. The aponeurosis and skin were sutured with a double suture. Omeprazole at a dose of 0.35 mg / kg was orally administered for 6 months.

Six months later, under the conditions of the surgical stage of nembutal anesthesia (40 mg / kg), blood was taken (1.5 ml), centrifuged (3000 rpm). The serum was then separated into cuvettes for determination of phosphorus, calcium, creatinine, albumin, alkaline phosphatase on an Olimpus analyzer. Then, bone tissue was collected in the area of the diaphysis of the femur and sternum and the level of calcium phosphorus was determined. Morphological examination of bone tissue was performed with preliminary decalcification. Tissue samples were fixed according to standard methods and stained with hematoxylin and eosin.

The results of studies of biochemical parameters in an animal with ovariectomy and oral administration of omeprazole showed that the level of calcium in the blood serum was 2.66 mmol / L (norm 2.16), alkaline phosphatase - 186.0 (norm 45.5), phosphorus - 1 , 92 (norm 1.6), albumin - 32.0% (norm 35), creatinine - 52.0 (norm 72). That is, there was an increase in the level of calcium, phosphorus and alkaline phosphatase, which indicates the development of osteoporosis.

The calcium content in bone tissue was 0.95 mg / g of tissue (normal 3.5 mg / g), phosphorus - 0.9 mg / g of tissue (normal 1.5 mg / g), which indicates a decrease in saturation of calcium and phosphorus bone tissue.

A morphological study revealed thinning of bone beams and degeneration of osteoblasts, expressed in a moderate degree.

The study was conducted on 18 rats in which a model of osteoporosis was reproduced. The purpose of the invention is to increase the accuracy of modeling of osteoporosis that occurs when exposed to proton pump inhibitors, is achieved.

Claims (1)

A method for modeling osteoporosis by administering a drug, characterized in that a bilateral oophorectomy is performed followed by the administration of omeprazole at a dose of 0.3-0.4 mg / kg orally for 6 months.