CN103877140A - Application of saussurea involucrate culture in preparation of product for promoting bone health - Google Patents

Abstract

The invention discloses the application of snow lotus culture in preparing products for promoting bone health, including the preparation of food or medicine for promoting bone health. The promotion of bone health is prevention and treatment of osteoporosis, improvement of bone calcium content, increase of bone density, improvement of rheumatoid arthritis and treatment of arthritis. Through animal tests and clinical tests, the present invention finds that the snow lotus culture can increase bone calcium content, increase bone density to prevent and treat osteoporosis, improve rheumatoid arthritis, and can be used in food and medicine for promoting bone health.

Description

Application of snow lotus culture in preparation of products promoting bone health

technical field

The invention relates to the technical fields of food and medicine, in particular to the application of snow lotus culture in preparing products for promoting bone health.

Background technique

Osteoporosis is a systemic bone disease characterized by low bone mass and damage to bone microarchitecture, resulting in increased bone fragility and susceptibility to fracture. It is a degenerative disease of the bones and the risk of developing it increases with age. With the extension of human life span and the arrival of aging society, osteoporosis has become an important health problem for all human beings. Osteoporosis is also a common and frequently-occurring disease that threatens menopausal women. Osteoporosis is caused by a variety of factors. Its basic pathological mechanism is the defect in the coupling of bone resorption and bone formation in the process of bone metabolism, which leads to the imbalance of calcium and phosphorus metabolism in the human body and the gradual decrease of bone density. Clinical symptoms. The most common symptom of osteoporosis is low back pain, accounting for 70% to 80% of pain patients. The pain spreads along the spine to both sides. The pain is relieved when lying on the back or sitting, and the pain is aggravated when standing upright or standing or sitting for a long time. The pain is mild during the day and aggravated at night and when waking up in the morning. Aggravated by straining to defecate.

Bone calcium is the content of calcium element in bones, and the reduction of bone calcium content is an important pathological feature of osteoporosis. The full name of bone density is bone mineral density. The level of bone mineral density is a measure of bone mass, reflects the degree of osteoporosis, and is an important basis for predicting the risk of fracture. Increasing the calcium content in bones and increasing bone density can prevent and treat osteoporosis.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by joint synovitis. The pathogenesis is very complicated and has not been fully elucidated so far. Its clinical manifestations are joint pain, swelling, poor flexion and extension, and morning stiffness. In RA, the persistent hyperplasia of the synovium and the destruction of cartilage and bone are its important pathological features. There are many animal models of RA, and the rat arthritis induced by Freund's complete adjuvant is one of the more classic models.

The effects of Saussurea sauraceae culture on improving bone calcium content, increasing bone density, preventing and treating osteoporosis have not been reported.

ZL201210213551.7 A traditional Chinese medicinal wine for the treatment of osteoporosis. It is recorded that "Tianshan snow lotus, Xihe willow, ground maple bark, stork grass, curculigo can invigorate the spleen and stomach, strengthen the spleen and lungs, and strengthen bones and muscles". "Tianshan Saussurea is used for rheumatoid arthritis, joint pain, lung cold cough, kidney and lower abdomen cold pain, excessive leucorrhea, etc.", there is no record that Saussurea or Saussurea culture can increase bone density, prevent and treat osteoporosis and related pharmaceuticals data.

Invention application 201310299562.6 A traditional Chinese medicine composition for the treatment of postmenopausal women with osteoporosis, records the traditional efficacy of snow lotus, "indications snow blindness, toothache, rheumatoid arthritis, impotence, irregular menstruation, red collapse, leucorrhea, gastric ulcer , hemorrhoids, bronchitis, heart disease, epistaxis and snake bites, etc., it has the functions of promoting menstruation and blood circulation, warming the uterus and dispelling stasis, dispelling cold and dehumidification, strengthening tendons and yang, stopping bleeding, and reducing swelling.” There is no record of snow lotus or Efficacy and related pharmaceutical data of Saussurea sauraceae culture in increasing bone density and preventing osteoporosis.

ZL200710017385.2 has the function of increasing bone density and its preparation and quality control method. Cimicifuga, the two synergistically improve bone mass and increase bone density" and "Calcium citrate, pearl powder and calcium carbonate are selected as calcium sources to prevent and alleviate bone loss to prevent fractures and maintain bone strength; at the same time, supplement casein Phosphopeptides promote better absorption of calcium by the human body”, there is no record of Saussurea or Saussurea cultures having the effect of increasing bone density and bone calcium and related pharmaceutical data.

In terms of anti-arthritis, the patent ZL201010519409.6 discloses that "the snow lotus culture composition has been found through experiments to be effective in preventing and alleviating arthritis", but the snow lotus culture composition given in this patent contains the following components: Snow lotus culture , cucumber seed powder, lettuce seed powder, celery seed powder, and calcium source, and the culture of snow lotus appeared to "make up for the singleness of rapeseed food on the market for arthritis health care", and did not mention the culture of snow lotus Independent role in arthritis. In the embodiment of anti-inflammatory effect, the animal model adopted is that acetic acid causes changes in the permeability of mouse abdominal capillaries and carrageenan causes swelling of the paws of rats, which can only illustrate that the above-mentioned composition has anti-inflammatory effect, but The role of Saussurea cultures in rheumatoid arthritis cannot be stated.

Contents of the invention

The purpose of the present invention is to propose the application of the snow lotus culture in the preparation of products for promoting bone health.

Application of snow lotus culture in preparation of products for promoting bone health.

The snow lotus culture of the present invention is obtained by selecting the snow lotus callus as the subculture seed and subcultured by cell culture technology, that is, the Chinese patent ZL200510115902.0 describes a method for large-scale culture of the snow lotus culture.

Further, the present invention provides the application of Saussurea edulis culture in the preparation of food or medicine for promoting bone health.

Further, the promotion of bone health is the prevention and treatment of osteoporosis.

Further, the promotion of bone health is to increase bone calcium content.

Further, the promotion of bone health is to increase bone density.

Further, the promotion of bone health is improving rheumatoid arthritis.

Further, the promotion of bone health is the treatment of arthritis.

Further, the drug is administered in the form of oral administration, injection, infusion or transdermal administration.

Further, the dosage of the food prepared by the Saussurea chinensis culture is 0.02-4g (calculated by the Saussurea chinensis culture) per day, preferably 0.1-0.2g.

Further, the dosage of the medicine prepared from the culture of snow lotus is 0.02-12 g per day (calculated based on the culture of snow lotus).

Invention effect:

Although the treatment of rheumatism and rheumatoid arthritis is one of the traditional efficacy of Saussurea involucrata, the culture of Saussurea involucrata in the present invention is obtained from Tianshan Saussurea indica through plant cell culture technology (see ZL200510115902.0 for specific cultivation methods), its color, The traits are completely different from Tianshan Saussurea; the composition and content of functional components are also very different from Tianshan Saussurea, the content of flavonoids, polysaccharides, chlorogenic acid, and syringin in the culture of Saussurea chinensis are much higher than that of wild Tianshan Saussurea. It can be considered that although the culture of snow lotus comes from Tianshan snow lotus, it is completely different from Tianshan snow lotus.

Through animal tests and clinical tests, the present invention finds that the snow lotus culture can increase bone calcium content, increase bone density to prevent and treat osteoporosis, improve rheumatoid arthritis, and can be used in food and medicine for promoting bone health.

Detailed ways

The following non-limiting examples can enable those skilled in the art to understand the present invention more fully, but do not limit the present invention in any way.

Example 1

BMD (Bone Mineral Density) is a commonly used physiological index in orthopedic clinics, and is currently the standard for diagnosing osteoporosis. The World Health Organization's definition of osteoporosis is based on the T value of bone density (comparing the bone density (BMD) obtained by the examination with the BMD of the normal young population to obtain a high (+) or low (-) young age The standard deviation (SD) number of people), as follows:

Normal: bone mineral density within 1 SD of the mean for young people (T value between +1 and -1);

Low bone density: the bone density is 1-2.5 SD lower than the average value of young people (T value is between -1 and -2.5);

Osteoporosis: bone mineral density 2.5 SD lower than the mean for young people (T value lower than -2.5);

Severe osteoporosis: Bone mineral density 2.5 SD below the mean for young adults (T-score below -2.5) with one or more osteoporotic fractures.

Sample: snow lotus culture capsules, each containing 100mg of snow lotus culture powder;

Dosage: two capsules per day;

Taking time: 6 months;

Bone Densitometer: Dual Energy X-ray Densitometer

Measurement site: distal radius

cases symptom T value before taking medicine T value after taking medicine Symptom improvement 1 back pain -3.1 -1.9 pain relief 2 low back pain -2.7 -0.6 pain gone 3 back pain -3.3 -1.5 pain relief 4 low back pain -2.5 -0.7 pain gone

Example 2

Saussurea cerevisiae culture used in clinical trials of rheumatoid arthritis

Test method: Case selection refers to the diagnostic criteria of rheumatoid arthritis of the American College of Rheumatology. The course of disease is 6 months to 11 years.

Number of cases: 15

Trial time: 4 weeks

Dosage: 2 times a day, 100mg sausage culture each time

Test results: Pain improved by 73.3%, swelling improved by 66.7%, and morning stiffness improved by 60.0%.

Comparison of the total effect of joint pain, swelling and morning stiffness in patients after treatment

case number number of painful joints curative effect case number number of swollen joints curative effect case number Morning stiffness curative effect pre-treatment after treatment pre-treatment after treatment pre-treatment after treatment 1 6 2 improve 1 6 4 improve 1 2 1 improve 2 6 2 improve 2 6 2 improve 2 2 2 invalid 3 4 0 disappear 3 4 0 disappear 3 2 1 improve 4 6 2 improve 4 6 2 improve 4 2 0 disappear 5 6 4 improve 5 6 6 improve 5 2 0 disappear 6 4 4 invalid 6 4 4 invalid 6 2 1 improve 7 4 4 invalid 7 4 4 invalid 7 2 1 improve

8 4 0 disappear 8 4 0 disappear 8 1 1 invalid 9 6 6 invalid 9 6 6 invalid 9 1 0 disappear 10 6 2 improve 10 6 2 improve 10 1 0 disappear 11 2 2 invalid 11 2 2 invalid 11 2 2 invalid 12 4 0 improve 12 4 0 improve 12 2 1 improve 13 2 0 disappear 13 2 0 disappear 13 2 2 invalid 14 4 0 disappear 14 4 4 invalid 14 1 1 invalid 15 4 0 disappear 15 4 0 disappear 15 2 2 invalid

Experimental example 1

Drug efficacy experiment of Saussurea edulis culture in increasing bone density, increasing bone calcium content, and preventing and treating osteoporosis

Materials and Methods

Test grouping: sham operation group, model control group, positive control group (estradiol 0.12mg/kg)

Test model: ovariectomized low bone density model

Experimental animals: SD adult female rats, adapted to feeding for 3-5 days before castration.

Modeling method: Rats underwent bilateral ovariectomy to castrate, and at the same time were given special feed to form a model with low bone density; another part of the rats underwent similar operations but did not remove bilateral ovaries, only part of the fat was removed, which was a sham model. surgery group.

The formula feed was formulated with reference to the AIN93 feed formula to prepare the semi-finished feed, and the soybean-derived ingredients in the feed were replaced with appropriate substances to avoid the interference of soybean isoflavones and other plant hormones on the experimental results. Formula composition: casein 23%, DL-methionine 0.3%, corn starch 32%, sucrose 30%, fiber 5%, corn oil 5%, mixed mineral salt 3.5%, mixed vitamin 1%, choline bitartrate 0.2%, Produced by Beijing Keao Xieli Feed Co., Ltd.

Snow lotus culture high-dose group (100mg/kg.d), snow lotus culture medium-dose group (33.3mg/kg.d), snow lotus culture low-dose group (16.7mg/kg.d) mg/kg of the present invention .d is the daily dose per kilogram of rat body weight.

Number of animals: 12 per group

Administration method: Oral gavage administration, once a day, for 3 months.

Test items: Three months later, the rat femur was taken to measure bone calcium, bone density (midpoint of femur and distal end of femur), ash weight and dry weight.

Determination of bone calcium: determination of bone calcium content by atomic absorption method;

Bone Densitometry: Measure the density of the midpoint of the femur and the distal end of the femur with a bone densitometer.

Effects of Snow Lotus Culture on Bone Calcium and Bone Density in Ovariectomized Rats

Note: Compared with the model group, *p<0.05, **p<0.01, ***p<0.001.

Results: After ovariectomy, compared with the sham operation group, the bone mineral density and bone calcium content of the rats in the model group were significantly reduced and had statistical significance, indicating that ovariectomized rats can cause bone calcium and The density decreased, and the modeling was successful; after 90 days of administration, the bone calcium content of each dose group of Saussurea chinensis culture was significantly higher than that of the model group, and there was statistical significance (p<0.01 or p<0.001); the bone of distal femur Compared with the model group, the density was significantly improved, with a significant difference (p<0.05 or p<0.01); the bone density of the midpoint of the femur also increased to varying degrees in each dose group, and the low-dose group had a significant difference compared with the model group (p<0.05).

Conclusion: Saussurea sauraceae culture can significantly increase bone calcium in rats with low bone density, and can also increase bone density at the midpoint of the femur and at the distal end of the femur. Therefore, the culture of snow lotus can be used to prevent and treat osteoporosis and promote bone health.

Experimental example 2

Rat arthritis model in complete Freund's adjuvant is one of the commonly used animal models for rheumatoid arthritis. The purpose of this experiment is to illustrate the therapeutic effect of Saussurea sauraceae culture on adjuvant arthritis in rats, and further illustrate its therapeutic effect on rheumatoid arthritis. Materials and Methods

Test groups: vehicle group (0.5% sodium carboxymethyl cellulose), positive control group (Xubi granule 2g/kg, indomethacin 4.5mg/kg), high-dose snow lotus culture group (140mg/kg.d) , medium dose group of Saussurea indica culture (70mg/kg.d), low dose group of Saussurea indica culture (35mg/kg.d)

Experimental model: Rat arthritis model in complete Freund's adjuvant

Test animals: rats with a body weight of 160-200 g, half male and half male.

Modeling method: Use a foot volume measuring instrument to measure the volume of the hind limbs on both sides of the rat, inject 0.1ml of Freund's complete adjuvant (containing 0.5 mg of inactivated Mycobacterium tuberculosis) subcutaneously into the right hind foot, and measure the animal's weight on the 15th day after injection , the volume of the hindlimbs on both sides, and the swelling rate and body weight of the left (non-inflammatory feet) hindlimbs were divided into six groups, with 10 rats in each group.

Number of animals: 10 per group

Administration method: intragastric administration, the administration volume is 20ml/kg. Once daily, give for seven days.

Test items and methods: The body weight of each animal and the volume of both hind limbs were measured on the 2nd, 5th, and 8th days after administration, and the swelling rate of the left and right paws was calculated.

Effects of Snow Lotus Culture on Inflamed Paw and Plantar Swelling in Rats with Adjuvant Arthritis

Note: Compared with the vehicle group, *p<0.05, Student’s-t test

The results showed that after the curative administration of the Saussurea sativa tissue cell culture, the 5th and 8th day, the Saussurea sauraceae tissue cell culture had a certain inhibitory effect on the inflammatory paw swelling.

Effects of Snow Lotus Culture on Paw Swelling of Non-inflammatory Rats with Adjuvant Arthritis

Note: Compared with vehicle group, *p<0.05, **p<0.01, Student’s-t test

The results showed that the 70mg/kg and 140mg/kg Saussurea tissue cell culture had a significant inhibitory effect on the swelling of non-inflammatory feet caused by delayed hypersensitivity on the 5th and 8th day after administration.

Conclusion: Therapeutic administration of Saussurea edulis tissue cell culture substances, 70mg/kg, 140mg/kg of Saussurea edulis tissue cell culture has a certain inhibitory effect on the inflammatory foot swelling rate of rats with adjuvant arthritis on the 5th and 8th day, but there is no statistics It has significant inhibitory effect on paw swelling caused by delayed hypersensitivity reaction in the face of the other antilimb (non-inflammatory foot).

Claims (10)

1. Application of Snow Lotus culture in the preparation of bone health promoting products. 2. The application according to claim 1, characterized in that the Saussurea edulis culture is used in the preparation of food or medicine for promoting bone health. 3. The application according to claim 1 or 2, characterized in that the promotion of bone health is the prevention and treatment of osteoporosis. 4. The application according to claim 1 or 2, characterized in that the promotion of bone health is to increase bone calcium content. 5. The application according to claim 1 or 2, characterized in that said promoting bone health is increasing bone density. 6. The application according to claim 1 or 2, characterized in that the promoting bone health is improving rheumatoid arthritis. 7. The application according to claim 1 or 2, characterized in that said promotion of bone health is treatment of arthritis. 8. The application according to any one of claims 1-7, characterized in that, the drug is administered in the form of oral administration, injection, infusion or transdermal administration. 9. The application according to any one of claims 1-7, characterized in that the dosage of the food prepared from the culture of snow lotus is 0.02-4 g per day. 10. The application according to any one of claims 1-7, characterized in that the dosage of the medicine prepared from the culture of snow lotus is 0.02-12 g per day.