KR100725916B1 - Composition for preventing bone loss comprising Salvia miltiorrhiza extracts and chitosan - Google Patents

Abstract

The present invention relates to the treatment and prevention of osteoporosis, and more particularly, to a composition for preventing bone loss, which comprises Salvia extract and chitosan.

According to the present invention, the composition of the present invention can be effectively used for the treatment or prevention of osteoporosis, and can be a plant extract functional raw material for bone health that suppresses bone loss of bone density and promotes calcium absorption into bone.

Description

Composition for preventing bone loss comprising Salvia miltiorrhiza extracts and chitosan

1 is a photograph of Salvia miltiorrhiza used in the present invention.

FIG. 2 shows different histological regions of the stunning tibia section in control (sham), ovarian resection (OVX) or OVX + SM mice.

The present invention relates to the treatment and prevention of osteoporosis, and more particularly, to a composition for preventing bone loss, which comprises Salvia extract and chitosan.

Osteoporosis is a condition in which bones that are normally hard cause genetic or acquired factors, such as menopause, aging, poor lifestyle (inadequate calcium intake, lack of exercise, lack of sunshine, smoking, excessive drinking, caffeine, etc.) and various osteoporosis. Diseases (gastrointestinal disease, liver disease, thyroid disease, parathyroid disease, hypersecretory hyperemia) and drugs (antacids, epilepsy, arthritis, constipation, corticosteroids, anticoagulants, antimedications) Minerals (particularly calcium) and substrates are diminished and eventually fractures can occur easily. Fractures usually occur in the wrists, ankles, ribs, spine, femur neck or ass bones.

For the treatment of osteoporosis, conventionally, female hormone, bisphosphonate, raloxifene, calcitonin, vitamin D active derivatives, etc., are administered as bone repression inhibitors, or fluorine, growth hormone, parathyroid hormone, etc. Other vitamin D metabolites, calcium and ifriflavones were administered.

Salvia miltiorrhiza is a perennial herb of the dicotyledon plant moth plant and belongs to Lamiaceae and is distributed in China, Korea (Gyeongbuk, Gangwon) and Japan. 40-80 cm in height. The whole is covered with yellowish white soft hairs and hairs. Root is long columnar, outer skin is scarlet. The leaves are opposite, single leaf or double pinnate leaf. Flowers bloom purple in May-June, run in layers, and long stamens come out. The medicinal herbs have short and rough roots and usually have stem marks on the apical tip. Root is long columnar, sometimes divided into 1 ~ 2 or several, 8 ~ 25㎝ long, 3 ~ 10㎜ in diameter. The outer surface is reddish brown to dark reddish brown with rough and vertical wrinkles. Figure 1 is a photograph of Dansam used in the present invention.

In Chinese medicine, Gi and blood (血) is not well communicated, if the blockage or wind and cold energy, moisture, etc. invaded the bones, various diseases come. Osteoporosis is the main cause of the inability to nourish bone due to damage to the kidneys. Here, regular kidneys refer to nutrients accumulated in the human body. Therefore, in order to eat well nutrition, avoid malnutrition due to overwork, and to communicate with blood donation regularly exercise, and also to take good communication with Gi and blood was taken. Its efficacy is described as surpassing that of Samul-tang. That means that the effect of promoting blood circulation is strong. Danshin is also a drug composed of tanshinone, mitiron, tanshinol, and vitamin E. It has the effect of vasodilation, antibacterial action, sedation and analgesic effect. Anti-cancer experiments have shown that it inhibits the respiration (energy metabolism) of cancer cells and inhibits the fermentation of glucose fermentation. In addition, Dansam is also used for cirrhosis of the liver, allergic purpura, coronary arteriosclerosis, and uterine disease (primary herb 本草綱目).

Thus, the inventors of the present invention, while searching for a new natural material that can prevent and treat osteoporosis, found that the composition containing the salvia extract and chitosan has a high bone loss prevention effect, to complete the present invention.

Accordingly, it is an object of the present invention to provide a composition for preventing bone loss useful for treating osteoporosis.

According to one aspect of the present invention, the present invention provides a composition for preventing bone loss, comprising Salvia miltiorrhiza extract as an active ingredient.

In the composition of the present invention, the extract may be extracted using water, an organic solvent or a mixed solvent thereof, which is a well-known extraction solvent for extracting conventional natural products. In the embodiment of the present invention, the hot water extraction method was used, but the organic solvent extraction method may be used by diluting methanol or ethanol in water at 50% to 100%.

In the composition of the present invention, preferably chitosan is further included as an active ingredient. When ginseng extract and chitosan were administered in combination, the effect of preventing bone loss was more significantly increased than when ginseng extract was administered alone.

In the composition of the present invention, although the salviae extract and chitosan may have any weight ratio that can be increased without inhibiting the effects of each other, preferably the salviae extract and chitosan in a weight ratio of 1: 1 to 5: 1 It is characterized by. Within the above weight ratio, the effects of Salvia extract and chitosan were sufficiently exhibited.

In the composition of the present invention, the composition for preventing bone loss is used for the treatment or prevention of osteoporosis. Since osteoporosis is caused by bone loss, preventing bone loss will effectively treat or prevent osteoporosis.

The risk factors for osteoporosis include women before and after menopause, women who have stopped menstruating for a long time, people who have thinner bones than others, dieters, people who have weakened bones due to old age, and fractures of their arms or legs. If you have plaster fixation, have a family history of osteoporosis, have excessive consumption of caffeine-containing beverages such as coffee, have excessive drinking, smoke, have had surgery for a fracture of your spine or limbs, or have been lying down for a long time due to other chronic diseases People who are nutritionally deficient or who eat less foods rich in calcium, such as dairy products and fresh vegetables. The composition of the present invention is also effective in the extraction and healing of bone grafts, implants, and extractions among children and adolescents and dental patients of growing age who want to be taller, such as nursing mothers, who need to be taller. Can be used.

In the present invention, the prophylactic effect of SM extract (SM) on the development of bone loss induced by ovarian resection (OVX) was studied in rats. We measured body weight and bone histomorphometry in sham, OVX or SM-administered OVX mice. From light microscopic analysis, a porous or erosive appearance was observed on the trabecular bone surface of the tibia of OVX mice, whereas the surface of the same bone of control rats and SM-administered mice consisted of fine particles. In OVX rats, spongy bone area and thickness were reduced by 50% compared to control rats, and these reductions were completely inhibited by SM administration for 7 weeks. In the present invention, the mechanical strength of the femur neck is remarkably improved by SM treatment for 7 weeks. In OVX rats, free T3 was normal in all cases, while free T4 increased significantly. Although there was no difference between OVX and SM-administered mice at the T3 level, a significant difference was found between them at the T4 level. These results show that the SM of the present invention can effectively prevent the development of bone loss induced by OVX in rats.

The composition of the present invention can be a plant extract functional raw material for bone health that suppresses bone loss of bone density and promotes calcium absorption into bone. The composition of the present invention has the function of promoting the proliferation and activity of osteoblasts, which make bones, and inhibiting the differentiation and activity of osteoclasts, which destroy bones, to prevent postmenopausal women and senile osteoporosis, and also to grow children and adolescents. It can be a food that helps develop. At present, Korea is the world's aging society. Aging social well-being is a top priority for bone health. Since the health of bone is an important issue directly related to the quality of life, the composition of the present invention will be a functional raw material to help improve the quality of life in an aging society and those suffering from bone.

In the present invention, the salviae extract shall include all of the extracts, fractions and purified products obtained in each step of the extraction, fractionation and purification treatment, diluents or concentrates thereof, or dried products thereof.

Salvia extract and chitosan, which are the raw materials of the present invention, may be formulated in the form of granules, tablets, capsules, or drinks by conventional methods known in the art. Moreover, in order to make storage and handling easy, you may add the carrier used for normal formulation, such as dextrin and cyclodextrin, and other arbitrary preparations.

In addition, it is suitable to administer soybean extract and / or chitosan of the present invention so that the daily intake of adult 1 ~ 3000mg. In addition, the dosage can be appropriately increased or decreased depending on age, symptoms, and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example 1: Preparation of Salvia Extract and Chitosan

Salvia extract is 15g of Salvia miltiorrhiza It was prepared by dissolving in 500 ml of distilled water for 3 hours at 60 ℃. The extracted salvia was filtered through a 0.45 mm filter and then lyophilized and stored at 4 ° C. Dansam was obtained by Kim.J from Oriental Medicine Hospital of Wonkwang University and College of Oriental Medicine, Wonkwang University. Validation data of Salvia ginseng extract was verified by the sample hall of Wonkwang University College of Pharmacy (No.99-02-0009). As chitosan, the water-soluble high molecular chitosan manufactured by Jagwang Co., Ltd. was used.

Example 2 Ovarian Removal Surgery and Administration of Chitosan Extract and Chitosan in Experimental Animals

50 females of 10-300g (10 animals in each group) were purchased from the Center for Experimental Animals. Animals were raised by circulation. The rats were prepared in individually standardized cages and fed a commercially available standard diet containing 1.2% calcium and 0.8% phosphorus. The 15-week-old rat underwent ovarian ablation (OVX) and laparotomy. Sodium nembutal was administered intraperitoneally, followed by anesthesia followed by removal of both ovaries. Comparative surgery was performed with the same stimulus to remove the ovaries to the outside. The ovaries removed were divided into four groups: oral administration of Salvia (30mg / ml), Chitosan (15mg / ml), Salvia (30mg / ml) + Chitosan (15mg / ml), and 2ml of saline solution. Seven weeks after drug administration, blood was collected and stored at -70 ° C until biochemical measurements were taken. The femur and tibia were dissected. The muscles attached to the right femur were removed cleanly and placed in sterile saline and stored at 4 ° C. for 8-16 hours. Twelve left femurs and eight randomly selected mice per group were fixed with Burkhart fixative and used by histological examination. All experiments were conducted using animals raised in accordance with the guidelines of animals used in the field of physiology and science published in the Korean Physiological Society, and were reared according to the guidelines that were approved and shipped by NIFSACC.

Example 3: Biochemical Analysis

Calcium, phosphorus, and alkaline phosphatase in serum were analyzed by laboratory standard techniques. Free T4 (FT4) and free T3 (FT3) in serum were measured with a Liso-Phase kit (Technogenetics, Milan, Italy) after separation of the hormonal chromatography layer using Sephadex LH-20 chromatography.

Example 4: Bone histomorphometry

The left femur was fixed for 24 hours with a Buckheart fixative for 24 hours and then cut in half with a diamond saw. Dehydrated with methanol and embedded with methyl methacrylate. Vertically cut femurs were cut to 4um thickness using Ao Autocut / Jung 1150 microtomes. For histological progression and histological analysis, cuts with bone cavity adjacent to the center of the femoral alley were selected. Sections cut to 4um thickness were stained according to Von cossa method for tetrachromium counterstaining in order to measure the reticular area of bone, the appearance of osteoblasts and the appearance of osteoclasts. Analysis of all bone tissues was done with the Bioquant Bone Morphometry System (R & M Biometrics Corp. Nashville, TN) described in previous literature. The measurement of the bone reticular tissue was further extended with the growth plate and the end of the bony trunk in the region starting from 1 mm adjacent to the femur to measure the junction of the femoral alley and the projection of the larger femur. Bone reticular area was measured at 200 magnifications, with the percentage of total reticular visual field versus the percentage of main tissue and osteoblast and osteoclast appearance. The quantity, width and distance of the fiber column were calculated.

Example 5 Measurement of Mechanical Strength of Femoral Head

The femoral head was cut at the rate of 0.095 radian / sec (5.43 ^o ) and cut by hydraulic experimental technique. First, the shaft fractured 19 mm above the knee joint bending to the anterior three points. Next, the neck was pulled down with the front of the laser in a straight line, with compression, flexion, rapid reproducibility, and head fracture. The load was measured by a mechanical test connecting the working hole through an amplifier to a microcomputer. Load / deviation curves were recorded using the connected Work Bench Mac software.

Experimental Results 1: Effect of Salvia Miltiorrhiza Extract (SM) and Chitosan (K) on Sponge Bone

Different histological regions of the stunning tibia section in control (sham), ovarian resection (OVX) or OVX + SM mice are shown in FIG. 1. As expected, loss of cavernous bone was observed in OVX mice and was most evident in the median bone media (FIG. 1). In SM-administered rats, areas partially filled with spongy bone (thick areas) were observed, showing a constant width of bone. In SM-treated rats, the spongy bone in the area between the growth junction and the thick portion was maintained compared to that of the control rats. In contrast, the loss of spongy bone in the region between the growth plate-mediate bone junction and the thick portion was markedly flexible in control rats. Spongy bone area (%) was measured (Table 1). As expected, the spongy bone area was significantly less in the OVX compared to the control. However, spongy bone area in SM-administered rats was maintained at the level observed in control rats, and spongy bone area was highest in SM + K-administered rats.

[Table 1] Effect of Salvia Miltiorrhizae Extract (SM) and Chitosan (K) on Sponge Bone Area (%) in OVX Rats

Values are Mean Standard Deviation. OVX: Ovariectomized. *: Statistical significance as compared with sham group (***: p 0.001). #: Statistical significance as compared with OVX group (#: p 0.05)

Experiment 2: Effect of Salvia Militiorrhizae (SM) and Chitosan (K) on Sponge Bone Thickness and Separation

Seven weeks after surgery, the spongy bone thickness of OVX rats was significantly less than that of the control group. The decrease in spongy bone thickness induced by OVX was inhibited by the administration of salvia extract for 7 weeks, and was most inhibited by the administration of salvia extract and chitosan (Table 2). The spongy bone separation of OVX rats was significantly inhibited by the administration of salvia extract for 7 weeks, and was most inhibited by the administration of salvia extract and chitosan (Table 3). In addition, there was no significant change in spongy bone marrow between OVX and SM-administered OVX mice (OVX + SM) (data not shown).

[Table 2] Effect of Salvia Miltiorrhizae (SM) and Chitosan (K) on the Sponge Bone Thickness (um) of OVX Rats

Values are Mean Standard Deviation. OVX: Ovariectomized. *: Statistical significance as compared with sham group (***: p 0.001). #: Statistical significance as compared with OVX group (###: p 0.01).

[Table 3] Effect of Salvia Militiorrhizae (SM) and Chitosan (K) on Sponge Bone Isolation (um) in OVX Rats

Values are Mean Standard Deviation. OVX: Ovariectomized. *: Statistical significance as compared with OVX group (***: p 0.001).

Experimental Results 3: Effect of Salviae Radix (SM) and Chitosan (K) on Osteoclast Number and Osteoblast Surface

The average number of osteoclasts on the surface of spongy bone and per unit area of spongy bone (mm ² ) is shown in Table 4 in three experimental groups. The number of osteoclasts per bone area was significantly affected by OVX. However, there was no significant change in osteoclast count between OVX mice and OVX + SM and OVX + SM + K mice. Although the surface of osteoblasts was larger in OVX mice, the effects induced by Salvia extract or Salvia extract and chitosan on osteoblast surface were not significant (Table 4).

[Table 4] Effect of Salmon Extract (SM) and Chitosan (K) on Osteoclast Number (No / mm) and Area (%) in OVX Rats

Values are Mean Standard Deviation. OVX: Ovariectomized. *: Statistical significance as compared with sham group (***: p 0.001).

Test Result 4: Effect of Salvia Militiorrhizae (SM) and Chitosan (K) on the Mechanical Strength of the Femoral Neck

OVX decreased the weight of ash in the femoral neck at each postoperative time point, and averaged 29.89 N in OVX group and 33.86 N in control group after 7 weeks of OVX. Maximum load on the femoral neck was also reduced due to estrogen deficiency at each time point after surgery (data not shown). The mechanical strength of the femoral neck was significantly improved by the treatment of salvia extract for 7 weeks, and most significantly by the treatment of salvia extract and chitosan (Table 5).

[Table 5] Effect of Saliva Extract (SM) and Chitosan (K) on the Mechanical Strength of the Femur Neck of OVX Rats

Values are Mean Standard Deviation. OVX: Ovariectomized. : Statistical significance as compared with sham group (*: p 0.05). Statistical significance as compared with OVX group (#: p 0.05).

Experimental Result 5: Effect of Salvia Miltiorrhiza Extract on Serum Biochemical Levels

In order to evaluate the effect of Salvia ginseng extract on the factors involved in anti-osteoporosis, serum biochemical analysis was performed. At week 7, serum biochemical measurements showed a marked increase in serum levels of alkaline phosphatase (ALP) or inorganic phosphorus (IP) in OVX mice compared to control (sham) mice. The increase in ALP was inhibited by the administration of Salvia extract. However, Salvia extract did not have a modulating effect on IP changes in OVX mice. However, serum calcium levels were not affected by administration of OVX or Salvia extract or Salvia extract and chitosan (Table 6). The data in Table 6 also mentions serum T3 levels and T4 levels. Thyrotoxicosis is associated with an increase in bone turnover leading to bone loss. The amount of T3 was not changed in OVX rats compared to control rats, but Mean T4 was significantly increased in OVX rats. When all the data for thyroid hormone measurements were evaluated, reduced levels of T4 were found in OVX rats administered with Salvia extract.

[Table 6] Effect of Salvia extract (SM) and chitosan (K) on serum biochemical levels of OVX rats

Values are Mean Standard Deviation. OVX: Ovariectomized. *: Statistical significance as compared with sham group (**: p 0.01; ***: p 0.001). #: Statistical significance as compared with OVX group (#: p 0.05; ##: p 0.01)

As described above, according to the present invention, the composition of the present invention can be effectively used for the treatment or prevention of osteoporosis, and will be a plant extract functional raw material for bone health that suppresses bone loss in bone density and promotes calcium absorption into bone. Can be.

Claims (5)

delete delete delete delete Danshen (Salvia miltiorrhiza) water soluble extract and chitosan 1: 1 to 5: The composition for the prevention and treatment of osteoporosis, comprising a weight ratio of 1.

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