CN116440182A - Traditional Chinese medicine composition for promoting height growth and preparation method and application thereof - Google Patents
Traditional Chinese medicine composition for promoting height growth and preparation method and application thereof Download PDFInfo
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- CN116440182A CN116440182A CN202310378269.2A CN202310378269A CN116440182A CN 116440182 A CN116440182 A CN 116440182A CN 202310378269 A CN202310378269 A CN 202310378269A CN 116440182 A CN116440182 A CN 116440182A
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/254—Acanthopanax or Eleutherococcus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Physical Education & Sports Medicine (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Rheumatology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a traditional Chinese medicine composition for promoting height growth, and a preparation method and application thereof, and belongs to the technical field of medicines. The composition provided by the invention consists of teasel root and acanthopanax root according to a mass ratio of 1-2:1-2, and the preparation method of the traditional Chinese medicine composition can be used for obtaining liquid medicine after decoction and filtration, or can be used for combining extracts of raw materials. The Chinese medicinal composition can express the increase of the thickness of the growth plate by increasing the proliferation rate of the chondrocyte proliferation region of the growth plate in a mode of increasing the cell number of the proliferation region, the pre-hypertrophy region and the hypertrophy region, thereby promoting the growth of the body.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a traditional Chinese medicine composition for promoting height growth, and a preparation method and application thereof.
Background
The dwarfism is characterized by that the incidence rate of the crowd is about 3% when the height is lower than the median 2 standard deviations (-2 SD) or the third percentile of the heights of children with the same sex and the same age, the etiology is complex, the differential diagnosis range is wide, and the replacement therapy of recombinant human growth hormone (rhGH) or combined gonadotropin-releasing hormone (GnRH) agonist is the most common clinical treatment measure for patients with the dwarfism. However, excessive growth hormone therapy may increase growth rate to cause rapid progress of puberty, combined GnRH agonist therapy often causes reduction of bone density, in addition, long-term growth hormone injection therapy also has a plurality of potential risks of disease occurrence, and growth hormone is expensive, inconvenient in injection use mode and large in individual variability of clinical treatment effect, so that development of the traditional Chinese medicine compound preparation with growth promotion potential is feasible and has huge prospects. At present, research on the compound medicament for treating the dwarfism at home and abroad is based on the dialectical treatment theory of traditional Chinese medicine, but lack of pharmacodynamic substance basis and molecular mechanism elucidation severely limits the application development of the compound medicament for the clinical treatment of the dwarfism.
Disclosure of Invention
In view of the above, the present invention aims to provide a height growth promoting traditional Chinese medicine composition, which can promote height growth by increasing the proliferation rate of chondrocytes in a proliferation region of a growth plate, and increasing the thickness of the growth plate in a manner of increasing the cell number in the proliferation region, the pre-hypertrophy region and the hypertrophy region.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a traditional Chinese medicine composition for promoting height growth, which consists of teasel root and acanthopanax root according to the mass ratio of 1-2:1-2.
The invention provides a preparation method of the traditional Chinese medicine composition, which comprises the steps of weighing raw materials according to the proportion of the raw materials, decocting and filtering to obtain liquid medicine.
The invention also provides a preparation method of the traditional Chinese medicine composition, the raw materials are weighed according to the proportion of the raw materials, the radix dipsaci extract and the acanthopanax extract are respectively extracted, and the extracts are mixed.
Preferably, the radix dipsaci extract and the acanthopanax extract are Chinese patent medicine particles.
Preferably, the concentration of the dipsacus saponin VI in the dipsacus extract is more than or equal to 2mg/mL, and the concentration of the dipsacus saponin B is more than or equal to 30 mug/mL; the concentration of the eugenol glycoside in the acanthopanax root extract is more than or equal to 50 mug/mL, and the concentration of the acanthopanax root glycoside E is more than or equal to 80 mug/mL.
The invention also provides application of the traditional Chinese medicine composition in preparing medicines for treating dwarfism.
Preferably, the Chinese medicinal composition promotes proliferation of cartilage cells of the tibial growth plate.
Preferably, the Chinese medicinal composition promotes the formation of bone trabeculae.
The invention also provides a medicine for treating dwarfism, which contains 0.1-100 wt% of the traditional Chinese medicine composition.
Preferably, the medicament comprises pharmaceutically acceptable excipients.
The invention has the beneficial effects that:
the traditional Chinese medicine composition can express the increase of the thickness of the growth plate by increasing the proliferation rate of chondrocytes in the proliferation area of the growth plate in a mode of increasing the cell number of the proliferation area, the pre-hypertrophy area and the hypertrophy area, thereby promoting the growth of the height. The traditional Chinese medicine composition can obviously increase the proliferation rate of ITS (insulin-transferrin-sodium selenite) -induced ATDC5 cells and promote the cartilage-like differentiation degree of ITS-induced ATDC5 cells.
Drawings
Fig. 1: the effects of different Chinese medicinal compatibility on the serum IGF-1, LH, FSH and E2 levels of rats in each group;
fig. 2: the different traditional Chinese medicine compatibility influences the development of the tibia growth plates of rats in each group;
fig. 3: the influence of different traditional Chinese medicine compatibility on the tibia trabeculae of rats in each group;
fig. 4: influence of teasel root-acanthopanax (XC) group on SD rat growth;
fig. 5: influence of radix Dipsaci-radix Acanthopanacis Senticosi (XC) group on ITS induced ATDC5 cell cartilage proliferation;
fig. 6: influence of radix Dipsaci-radix Acanthopanacis Senticosi (XC) group on ITS induced ATDC5 cell cartilage differentiation;
fig. 7: sox9, col2a1, acan, col10a1 gene expression levels during ATDC5 cell proliferation and differentiation;
fig. 8: determination of the content of the major active ingredients of the teasel root-acanthopanax (XC) group.
Detailed Description
The invention provides a traditional Chinese medicine composition for promoting height growth, which consists of teasel root and acanthopanax root according to the mass ratio of 1-2:1-2 of raw medicinal materials, wherein the mass ratio is preferably 1:1.
Teasel root, chinese medicine name. Is dry root of Dipsacus asperWall. Ex Henry of Dipsacus of Dipsacaceae. Has effects of invigorating liver and kidney, strengthening tendons and bones, and promoting blood circulation. Is mainly used for treating liver and kidney deficiency, soreness of waist and knees, cold-dampness arthralgia, metrorrhagia, bleeding, fetal irritability, traumatic injury, fracture of bones and muscles, etc.
Acanthopanax root and its Chinese medicine name. Is rhizome or stem of Acanthopanax senticosus (Rupr. Etmaxim) Harms belonging to Acanthopanax of Araliaceae. Has effects of invigorating qi, invigorating spleen, tonifying kidney, and tranquilizing. Is mainly used for treating spleen and lung qi deficiency, kidney deficiency, soreness of waist and knees, heart and spleen deficiency, insomnia, amnesia and the like.
The invention provides a preparation method of the traditional Chinese medicine composition, which comprises the following steps: the first method comprises the steps of weighing the raw materials according to the ratio of the raw materials, decocting and filtering to obtain liquid medicine; as an alternative implementation mode, the invention weighs the raw medicinal materials of dipsacus root and acanthopanax, 15g of raw medicinal material powder is weighed and mixed after being processed by a traditional Chinese medicine pulverizer, 1.8L of distilled water is added according to the ratio of feed liquid to liquid of 1:60, and the mixture is decocted for 1.5 hours at 100 ℃, and the combined medicinal liquid of dipsacus root and acanthopanax is prepared after filtering by a filter screen for standby.
The second method comprises weighing the raw materials according to the ratio of the raw materials, respectively extracting to obtain radix Dipsaci extract and radix Acanthopanacis Senticosi extract, and mixing the extracts; in an alternative implementation mode, the preparation method comprises the steps of taking radix dipsaci and acanthopanax root raw medicinal materials, respectively treating the raw medicinal materials by a traditional Chinese medicine pulverizer, weighing raw medicinal material powder, adding 300mL of distilled water according to a ratio of feed liquid to liquid of 1:20, extracting for 1h at the water bath 80 ℃ under the conditions of ultrasonic power of 100W and ultrasonic frequency of 40KHz, and filtering by a filter screen to obtain radix dipsaci extract or acanthopanax root extract; mixing radix Dipsaci extract and radix Acanthopanacis Senticosi extract for use.
Preferably, the teasel root extract and the acanthopanax root extract are Chinese patent medicine particles. As an optional implementation mode, the invention uses the teasel root and acanthopanax root Chinese patent medicine particles together dissolved in double distilled water.
The teasel root extract requires that the concentration of teasel root saponin VI is more than or equal to 2mg/mL, preferably, the concentration of teasel root saponin VI is 2.5-5 mg/mL; the concentration of the teasel root saponin B is required to be more than or equal to 30 mug/mL, preferably, the concentration of the teasel root saponin B is 35-60 mug/mL; the concentration of the eugenol glycoside in the acanthopanax root extract is more than or equal to 50 mug/mL, preferably, the concentration of the eugenol glycoside is 55-90 mug/mL; the concentration of the eleutheroside E is required to be more than or equal to 80 mug/mL, and the preferred concentration of the eleutheroside E is 85-110 mug/mL.
The invention also provides application of the traditional Chinese medicine composition in preparing medicines for treating dwarfism. The traditional Chinese medicine composition promotes proliferation of cartilage cells of the tibia growth plate and promotes formation of bone trabeculae.
The invention also provides a medicine for treating dwarfism, which contains 0.1-100 wt% of the traditional Chinese medicine composition. The medicine also comprises pharmaceutically acceptable auxiliary materials, and the invention does not limit specific auxiliary materials.
As an alternative embodiment, the content of the effective components of the medicament for treating the dwarfism is calculated according to the administration of 35-45 g crude drugs by 40kg of clinical children, preferably 40-41 g crude drugs by 40kg of clinical children.
The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the following examples, the experimental animals were:
Sprague-Dawley (SD) rats weighing 200-250 g purchased from Jinan Pengyue laboratory animals Breeding Limited (license number: SCXK (robust) 20190003) and fed and drunk freely.
In the following examples, conventional methods are used unless otherwise specified.
Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1
A traditional Chinese medicine composition for promoting height growth comprises radix dipsaci and acanthopanax according to a raw medicinal material mass ratio of 1:1. The preparation method comprises the following steps: weighing 20g of each of the raw medicinal materials of radix dipsaci and acanthopanax, processing by a traditional Chinese medicine pulverizer, weighing 15g of each of the raw medicinal material powders, mixing, adding 1.8L of distilled water according to the ratio of feed liquid to liquid of 1:60, decocting at 100 ℃ for 1.5h, and filtering by a filter screen to obtain a radix dipsaci and acanthopanax combined liquid medicine for standby.
Example 2
A traditional Chinese medicine composition for promoting height growth comprises a teasel root granule preparation and an acanthopanax root granule preparation according to the mass ratio of raw medicinal materials of 1:1. The preparation method comprises the following steps: weighing 15g of raw medicinal material powder after being processed by a traditional Chinese medicine pulverizer, adding 300mL of distilled water according to the ratio of feed liquid to liquid of 1:20, extracting for 1h in water bath at 80 ℃ under the conditions of ultrasonic power of 100W and ultrasonic frequency of 40KHz, and filtering by a filter screen to obtain a teasel root extract; taking acanthopanax root raw medicinal material, treating the acanthopanax root raw medicinal material by a traditional Chinese medicine pulverizer, weighing 15g of raw medicinal material powder, adding 300mL of distilled water according to the ratio of feed to liquid of 1:20, extracting for 1h in a water bath at 80 ℃ under the conditions of ultrasonic power of 100W and ultrasonic frequency of 40KHz, and filtering by a filter screen to obtain acanthopanax root extract; mixing radix Dipsaci extract and radix Acanthopanacis Senticosi extract for use.
Example 3
A traditional Chinese medicine composition for promoting height growth comprises a teasel root granule preparation and an acanthopanax root granule preparation according to the mass ratio of raw medicinal materials of 1:1. The preparation method comprises the following steps: the teasel root and the acanthopanax are granular preparations, the teasel root and the acanthopanax are purchased from Jiangyin Tianjiang pharmaceutical industry Co., ltd, the mass of raw medicinal materials of each pack of granular preparations is 15g, the concentration ratio of the teasel root to the acanthopanax granules is 4.95 times and 10.12 times respectively, and the dosage of each single medicinal material is 1.5625g/kg, 1.2624g teasel root and 0.6176g acanthopanax are respectively weighed according to the ratio of 1:1 and dissolved in 40mL double distilled water for standby.
Comparative example 1
A Chinese medicinal composition comprises radix Codonopsis and radix Acanthopanacis Senticosi at a mass ratio of 1:1. The preparation method comprises the following steps: the radix codonopsis and the acanthopanax are granular preparations, the raw medicinal materials of each pack of granular preparations are 15g in mass, the concentration ratio of the radix codonopsis and the acanthopanax is 3.44 times and 10.12 times respectively, and each single medicinal material dose is 1.5625g/kg, 1.816g of radix codonopsis and 0.6176g of acanthopanax are respectively weighed according to the ratio of 1:1 and dissolved in 40mL of double distilled water for standby.
Comparative example 2
A Chinese medicinal composition comprises radix Codonopsis and radix Dipsaci at a mass ratio of 1:1. The preparation method comprises the following steps: the radix codonopsis and the radix dipsaci are granular preparations, purchased from Jiangyin Tianjiang pharmaceutical industry Co., ltd, the mass of raw medicinal materials of each pack of granular preparations is 15g, the concentration ratio of the radix codonopsis and the radix dipsaci granules is 3.44 times and 4.95 times respectively, and each single medicinal dosage is 1.5625g/kg, 1.816g of radix codonopsis and 1.2624g of radix dipsaci are respectively weighed according to the ratio of 1:1 and dissolved in 40mL double distilled water for standby.
Comparative example 3
A Chinese medicinal composition comprises radix Codonopsis, radix Dipsaci and radix Acanthopanacis Senticosi at a mass ratio of 1:1:1. The preparation method comprises the following steps: the radix codonopsis, the radix dipsaci and the acanthopanax are all granular preparations, the raw medicinal materials of each pack of granular preparations are 15g, the concentration ratio of the radix codonopsis, the radix dipsaci and the acanthopanax granules is 3.44 times, 4.95 times and 10.12 times respectively, and each single medicinal material dose is 1.5625g/kg, 1.816g of radix codonopsis, 1.2624g of radix dipsaci and 0.6176g of acanthopanax are respectively weighed according to the ratio of 1:1:1 and dissolved in 40mL double distilled water for standby.
Comparative example 4
A Chinese medicinal composition comprises radix Codonopsis, radix Dipsaci, radix Acanthopanacis Senticosi and radix Paeoniae Rubra at a mass ratio of 1:1:1:1. The preparation method comprises the following steps: the radix codonopsis, the radix dipsaci, the radix acanthopanacis and the radix paeoniae rubra are all granular preparations, the raw medicinal materials of each pack of granular preparations are 15g, the concentration ratio of the radix codonopsis, the radix dipsaci, the radix acanthopanacis and the radix paeoniae rubra granular preparation is 3.44 times, 4.95 times, 10.12 times and 7.16 times respectively, and each single medicinal material dose is 1.5625g/kg, 1.816g of radix codonopsis, 1.2624g of radix dipsaci, 0.6176g of radix acanthopanacis and 0.8728g of radix paeoniae rubra are respectively weighed according to the ratio of 1:1:1:1 and dissolved in 40mL double distilled water for standby.
Example 4
1. The experimental method comprises the following steps: SD rats (healthy rats) are animal subjects, and the traditional Chinese medicine compositions of the example 3 and the comparative examples 1 to 4 are subjected to single factor control experiments after being converted according to clinical dosage standards (calculated by 40g crude drugs given according to 40kg clinical children medication, converted into rat medication doses of 6.25g/kg, and executed according to 1mL/100g standard):
control group (Control): the stomach is irrigated with double distilled water in the same amount as the administration group.
Radix dipsaci-acanthopanax (XC): the Chinese medicinal composition described in example 3.
Radix codonopsis pilosulae-acanthopanax (DC): a Chinese medicinal composition according to comparative example 1.
Radix codonopsis-radix Dipsaci (DX): the traditional Chinese medicine composition described in comparative example 2.
Radix codonopsis pilosulae-radix dipsaci-acanthopanax (DXC): the Chinese medicinal composition described in comparative example 3.
Radix codonopsis pilosulae-radix dipsaci-acanthopanax root-red paeony root (DXCC): the Chinese medicinal composition of comparative example 4.
SD rats are respectively administrated with radix Dipsaci-radix Acanthopanacis Senticosi (XC), radix Codonopsis-radix Acanthopanacis Senticosi (DC), radix Codonopsis-radix Dipsaci (DX), radix Codonopsis-radix Dipsaci-radix Acanthopanacis Senticosi (DXC), radix Codonopsis-radix Dipsaci-radix Acanthopanacis Senticosi-radix Paeoniae Rubra (DXCC) group medicines by continuous intragastric administration for 4 weeks; each group was given 1 time per day by gavage, each dose being 1mL/100g.
2. After 4 weeks of dosing, serum biochemical index and tibia of rats were tested for each group:
the results of comparing the biochemical index of rat serum (IGF-1, LH, FSH, E2) between groups are shown in FIG. 1. As can be seen from fig. 1, radix dipsaci-acanthopanax (XC), radix codonopsis-acanthopanax (DC), radix codonopsis-radix Dipsaci (DX), radix codonopsis-radix dipsaci-acanthopanax (DXC), radix codonopsis-radix dipsaci-acanthopanax-radix paeoniae rubra (DXCC) all showed a trend of increasing serum IGF-1 level and inhibiting FSH, LH level of rats, and no significant difference (p > 0.05) was shown between groups.
The results of comparing the growth plate development of the rat tibia between the groups are shown in figure 2. As can be seen from fig. 2, codonopsis pilosula-acanthopanax (DC) and teasel root-acanthopanax (XC) can significantly promote the endochondral ossification process of the proximal metaphyseal growth plate of the tibia of the rat compared with the control group.
The tibial bone trabecular morphology of each group of rats was compared and the results are shown in fig. 3. As can be seen from fig. 3, teasel root-acanthopanax (XC) can significantly promote the formation of bone trabeculae compared with the other groups.
The results shown in FIGS. 1-3 indicate that radix Dipsaci-radix Acanthopanacis Senticosi (XC) has growth promoting potential.
3. The gastric lavage administration time of radix Dipsaci-radix Acanthopanacis Senticosi (XC) group and Control group (Control) is prolonged to 6 weeks.
After 6 weeks, the effect of teasel root-acanthopanax (XC) administration group on SD rat growth was investigated, and the results are shown in fig. 4. As can be seen from fig. 4, the weight and body length of the rats in the XC combination preparation administration group showed a certain increase compared to the normal control group, but there was no statistical significance (p > 0.05), and although there was no significant change in body length and weight, the number of chondrocytes in the proliferation zone, the pre-hypertrophy zone and the hypertrophy zone of the large tibial growth plate in the XC treatment group was significantly increased, and the thickness of the growth plate was significantly increased, demonstrating that the dipsacus root-acanthopanax root composition has significant growth-promoting potential.
4. Analysis of the effect of XC on ITS induction of ATDC5 cell chondroid differentiation:
ITS-induced ATDC5 cells were used as an in vitro model to simulate growth plate endochondral ossification in vitro.
1.2624g of teasel root and 0.6176g of acanthopanax are weighed and dissolved in 40mL of double distilled water to prepare XC solution with animal experimental action dose in vivo for later use. The XC is subjected to 20-fold, 100-fold, 500-fold, 1000-fold and 2000-fold gradient dilution by using a DMEM-Ham' sF12 cell culture medium, the action dosage range of an XC cell experiment is preliminarily determined to be 100-1000-fold by adopting CCK-8 to detect the cell activity, and the cell action dosage of the teasel root-acanthopanax combination preparation is further designed to be 125-1000-fold dilution: the low dose group (LD) was diluted 1000-fold; medium dose group (MD) was 500-fold diluted; high dose group (HD) was 250-fold diluted; very high dose group (VHD) was 125-fold diluted.
The effect of XC combination preparation on ITS induced cartilage proliferation of ATDC5 cells was investigated by CCK-8 and crystal violet staining methods, and the results are shown in FIG. 5. The experimental results of CCK-8 in FIG. 5 prove that XC obviously increases proliferation of ITS induced ATDC5 cells in a concentration-dependent manner, and the promotion effect of the XC combined preparation on proliferation of ITS induced ATDC5 cells can be obviously observed by adopting crystal violet staining on the 7 th day of induction, which indicates that the proliferation rate of ITS induced ATDC5 cells can be obviously increased by treating (125-fold dilution) the teasel root-acanthopanax combined preparation.
The effect of XC combination preparations on ITS induced chondrodifferentiation of ATDC5 cells was investigated using aliskiren blue and alizarin red staining on ATDC5 cells, and the results are shown in fig. 6. The results of the alisxin blue staining and alizarin red staining in fig. 6 demonstrate that the staining of the combination treatment of ITS and XC (very high dose) ATDC5 cells for 7 days, 14 days and 21 days is significantly deeper than that of the induction of ITS alone, demonstrating that the secretion of proteoglycan and calcium nodule formation of the ATDC5 cells after XC treatment are significantly increased, and therefore, XC can significantly promote the chondroid differentiation of the ITS-induced ATDC5 cells at 21 days.
Total RNA samples of each group of cells on day 0, day 7, day 14 and day 21 are respectively extracted by adopting a Trizol method, are subjected to reverse transcription into cDNA, and are subjected to detection and analysis on changes of expression levels of marker genes such as Sox9, col2a1, acan, col10a1 and the like in the process of inducing ATDC5 cell cartilage-like differentiation by ITS by using SYBRGreenqPCR by taking GAPDH as an internal reference gene, and the results are shown in FIG. 7. It was found that XC (very high dose) treatment did not affect the mRNA expression level of marker genes during 21 days of differentiation of cells, suggesting that the dipsacus root-acanthopanax combination preparation may exhibit an increase in growth plate thickness by increasing the proliferation rate of chondrocyte proliferation regions of growth plate in such a way as to increase the cell number of the regions, pre-hypertrophic regions and hypertrophic regions, thereby achieving the potential for promoting growth.
Example 5
Measuring the concentrations of the eugenol glycoside and the eleutheroside E in the teasel root-eleutheroside E combined preparation in the embodiment 3 by adopting an RP-HPLC method, respectively establishing standard curves by using eugenol glycoside and eleutheroside E standard substances, weighing a proper amount of uniformly mixed samples, putting the samples into a conical flask, passing through a 0.45 mu m microporous filter membrane, and measuring the concentrations of the eugenol glycoside and the eleutheroside E by using an upper machine, wherein the concentrations are 67.70 mu g/mL and 94.25 mu g/mL; and (3) measuring the concentration of the dipsacus root saponin VI and the dipsacus root saponin B in the dipsacus root-acanthopanax root combined preparation by adopting a UPLC-MS/MS method, respectively establishing standard curves by using the dipsacus root saponin VI and the dipsacus root saponin B standard substances, accurately weighing a proper amount of samples in a centrifuge tube, adding water into the centrifuge tube, vortex mixing uniformly, ultrasonically extracting and centrifuging, and then, loading the supernatant after passing through a 0.22 mu m microporous filter membrane to detect the concentration of the dipsacus root saponin VI and the dipsacus root saponin B, wherein the concentration is 45.35 mu g/mL and 3.99mg/mL respectively, and the result is shown in figure 8. As shown in FIG. 8, in the teasel root-acanthopanax root composite preparation in example 3, the concentration of teasel root saponin VI is more than or equal to 2mg/mL, the concentration of teasel root saponin B is more than or equal to 30 mug/mL, the concentration of syringin is more than or equal to 50 mug/mL, and the concentration of acanthopanax root saponin E is more than or equal to 80 mug/mL.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. The traditional Chinese medicine composition for promoting height growth is characterized by comprising teasel root and acanthopanax root according to the mass ratio of 1-2:1-2.
2. The method for preparing the traditional Chinese medicine composition according to claim 1, wherein the raw materials are weighed according to the proportion, and the liquid medicine is obtained after decoction and filtration.
3. The method for preparing the traditional Chinese medicine composition according to claim 1, wherein the raw materials are weighed according to the proportion of the raw materials, the radix dipsaci extract and the acanthopanax extract are respectively obtained by extraction, and the extracts are mixed.
4. The preparation method of claim 3, wherein the teasel root extract and the acanthopanax root extract are Chinese patent medicine particles.
5. The preparation method according to any one of claims 3 to 4, wherein the concentration of dipsacus saponin VI in the dipsacus extract is not less than 2mg/mL, and the concentration of dipsacus saponin B is not less than 30 μg/mL; the concentration of the eugenol glycoside in the acanthopanax root extract is more than or equal to 50 mug/mL, and the concentration of the acanthopanax root glycoside E is more than or equal to 80 mug/mL.
6. The use of the Chinese medicinal composition according to claim 1 in the preparation of a medicament for treating dwarfism.
7. The use of claim 6, wherein the Chinese medicinal composition promotes proliferation of tibial growth plate chondrocytes.
8. The use according to claim 6, wherein the Chinese medicinal composition promotes the formation of bone trabeculae.
9. A medicament for treating dwarfism, which is characterized by comprising 0.1-100 wt% of the traditional Chinese medicine composition of claim 1.
10. The medicament of claim 9, wherein the medicament comprises pharmaceutically acceptable excipients.
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