Application of American ginseng stem and leaf total saponins in treatment of postmenopausal osteoporosis
By using total saponin of American ginseng stems and leaves as a drug component, the adverse reactions and high cost problems in postmenopausal osteoporosis treatment were solved, and effective osteoporosis treatment and prevention were achieved without adverse reactions.
Patent Information
- Application Number
- CN202411715621.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-27
- Publication Date
- 2025-02-21
AI Technical Summary
Osteoporosis caused by the decline in estrogen levels after menopause, existing treatment methods have adverse reactions and high costs.
The total saponin of American ginseng stem and leaf is used as a pharmaceutical ingredient and is administered in the form of oral preparations. The total saponin of American ginseng stem and leaf includes ginseng saponin Rd, Rb3, Rb2, F11 and other ingredients. It has gentle pharmacological effects and no adverse reactions. It is suitable for long-term use.
Total saponin of American ginseng stems and leaves can effectively reduce postmenopausal osteoporosis, improve bone density, improve bone metabolism, reduce fracture risk, and have no adverse reactions. It is suitable for long-term use.
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Figure CN119488539A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of biomedicine, and in particular to application of total saponins from American ginseng stems and leaves in treating postmenopausal osteoporosis. Background Art
[0002] Postmenopausal osteoporosis is a common and frequently occurring disease in postmenopausal women. Modern medicine believes that postmenopausal osteoporosis is a high-conversion metabolic disease caused by an imbalance between bone resorption by osteoclasts and bone formation by osteoblasts. It has decreased bone mass and destruction of bone microstructure as its main pathological features, and systemic bone pain and easy fractures as its clinical characteristics.
[0003] Estrogen is one of the most important steroid hormones in women, mainly secreted by the ovaries. As women age, ovarian function declines, estrogen levels decrease, and the associated bone metabolism abnormalities can eventually lead to postmenopausal osteoporosis. Human bones are always in a dynamic remodeling process, which requires the synergistic action of two cell types, osteoclasts and osteoblasts. Estrogen can regulate the state of bones by affecting the bone remodeling rate. Estrogen can directly induce osteoclast apoptosis and reduce in vitro induced osteoclast differentiation, thereby reducing the bone resorption of osteoclasts. The significant decrease in estrogen levels after menopause not only weakens the effect on osteoclasts, leading to trabecular perforation and loss of connectivity; it also reduces the effect on osteoblasts, leading to bone loss, and the width of the bone wall begins to slowly decrease. In addition, the decline in estrogen leads to an increase in bone turnover rate, especially when the bone resorption rate is significantly greater than the bone formation rate. This imbalance leads to the loss of trabecular connectivity and cortical bone porosity, which leads to increased bone fragility and fracture susceptibility.
[0004] Given that the significant decline in estrogen levels after menopause significantly affects the occurrence and development of osteoporosis in women, some research has been conducted on its prevention and treatment measures. Modern medicine mainly adopts drug therapy and basic intervention to treat postmenopausal osteoporosis. While Western medicine can relieve patients' pain, increase bone density, and improve bone metabolism, long-term use of Western medicine still has many adverse reactions and high costs, which brings certain difficulties to clinical treatment. In particular, the use of hormone replacement therapy to prevent and treat osteoporosis has achieved certain clinical effects, but as time goes by, it increases the risk of cardiovascular events, breast cancer, and endometrial cancer.
[0005] In view of this, in order to solve the deficiencies of the prior art, the present invention provides the use of total saponins from American ginseng stems and leaves in the treatment of postmenopausal osteoporosis, which has a good research and development prospect. Summary of the invention
[0006] The purpose of the present invention is to provide an application of total saponins from stems and leaves of American ginseng in treating postmenopausal osteoporosis. The total saponins from stems and leaves of American ginseng have mild effects, no adverse reactions, and can be used sustainably, and have broad medical prospects and economic value.
[0007] To achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:
[0008] The present invention provides application of total saponins of American ginseng stems and leaves in preparing a medicine for treating postmenopausal osteoporosis.
[0009] Preferably, the total saponins from American ginseng stems and leaves include ginsenoside Rd, ginsenoside Rb3, ginsenoside Rb2 and pseudo-ginsenoside F11.
[0010] Further preferably, the total saponins from American ginseng stems and leaves also include ginsenoside Rc, ginsenoside Re, ginsenoside Rg1, ginsenoside Rg2, ginsenoside Rg3 and ginsenoside F2.
[0011] Preferably, the postmenopausal osteoporosis is caused by postmenopausal estrogen deficiency.
[0012] Preferably, the postmenopausal osteoporosis includes early postmenopausal osteoporosis and late postmenopausal osteoporosis.
[0013] Preferably, the symptoms of postmenopausal osteoporosis include hunchback, shortened stature and bone pain.
[0014] Further preferably, the symptoms of postmenopausal osteoporosis also include local tenderness and percussion pain.
[0015] Preferably, the complications of postmenopausal osteoporosis include bone fractures, chest tightness, shortness of breath, cough, abdominal distension and constipation.
[0016] Further preferably, the complications of postmenopausal osteoporosis include fractures and chest tightness.
[0017] Preferably, the medicine contains an effective amount of total saponins from American ginseng stems and leaves.
[0018] Further preferably, the mass proportion of total saponins from stems and leaves of American ginseng in the medicine is 16.7%.
[0019] Further preferably, the drug is in the form of at least one selected from pills, capsules, granules, oral liquids, powders, tablets, lozenges, and lozenges, and a suitable drug carrier in the art may be selected for different dosage forms.
[0020] The pharmaceutical carrier used can be solid, liquid or gas. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil and water. Examples of gaseous carriers include carbon dioxide and nitrogen.
[0021] When preparing the composition for oral dosage form, any convenient pharmaceutical medium can be used. For example, water, ethanol, oil, alcohol, flavoring agent, preservative, coloring agent, etc. can be used to form oral liquid preparations, such as suspensions and solutions; while carriers, such as starch, sugars, microcrystalline cellulose, diluents, granulating agents, emulsifiers, lubricants, binders, disintegrants can be used to form oral solid preparations, such as powders, capsules and tablets. Tablets and capsules are the preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques.
[0022] The pharmaceutical composition suitable for parenteral administration of the present invention can be prepared as an aqueous solution or suspension of the active compound. Suitable surfactants such as hydroxypropylcellulose can be included. Dispersions can also be prepared in glycerol, liquid polyethylene glycol and oil mixtures thereof. In addition, preservatives can be added to prevent the harmful growth of microorganisms.
[0023] The pharmaceutical preparation of the present invention may be in a form suitable for topical use, such as an aerosol, cream, ointment, lotion, powder or the like.
[0024] The pharmaceutical preparation of the present invention can be in a form suitable for rectal administration, wherein the carrier is a solid. Preferably, the mixture is made into a unit dose suppository. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be prepared by first forming a composition containing a softened or melted carrier, followed by cooling and shaping in a mold.
[0025] In addition to the above-mentioned carrier components, the above-mentioned pharmaceutical preparations may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. In addition, other excipients, such as colorants and flavoring agents such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc., may be added.
[0026] More preferably, the drug is an oral pharmaceutical preparation.
[0027] The beneficial effects of the present invention are:
[0028] Animal experiments have confirmed that total saponins from American ginseng stems and leaves can reduce osteoporosis in postmenopausal mice, indicating that total saponins from American ginseng stems and leaves can effectively treat osteoporosis caused by postmenopausal estrogen deficiency. BRIEF DESCRIPTION OF THE DRAWINGS
[0029] Figure 1 To investigate the effects of total saponins from American ginseng stems and leaves on the body weight of high-fat-fed and castrated mice.
[0030] Figure 2 To investigate the effects of total saponins from American ginseng stems and leaves on bone mass and bone microarchitecture in high-fat-fed and castrated mice, Micro-CT was used to detect trabecular volume fraction (BV / TV), average trabecular thickness (Tb.Th) and bone mineral density (BMD) at the bottom of the growth plate on the knee joint side of the distal femur of mice. Scale: 500 μm.
[0031] Figure 3 To investigate the effects of total saponins from American ginseng stems and leaves on the expression of PINP (bone formation) and CTX-1 (bone resorption) in the serum of osteoporotic mice induced by high-fat diet and castration.
[0032] Figure 4 This is the effect of total saponins from American ginseng stems and leaves on the histopathological findings of osteoporotic mice fed a high-fat diet combined with castration. Conventional HE staining: scale bar: 50μm.
[0033] Figure 5 This is the effect of total saponins from American ginseng stems and leaves on the histopathological findings of osteoporotic mice induced by high-fat diet and castration, including TRAP staining of femur: scale bar: 50μm. DETAILED DESCRIPTION
[0034] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is only an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the invention of the present invention based on the disclosed content, and they should also fall within the scope of the present application.
[0035] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all weight contents. Unless otherwise specified, it is understood that the process is carried out at room temperature.
[0036] Example 1
[0037] The main component of the total saponin of American ginseng stems and leaves used in the present invention is the clinically used drug Xinyue Capsule (national drug license number: No. Z20030073).
[0038] Results test:
[0039] 1. Materials
[0040] 1.1 Animals
[0041] SPF 8-week-old ApoE in C57BL / 6J background - / - 75 female mice and 10 C57BL / 6J female mice were housed in cages in a breeding room with natural light supplemented by artificial light at a temperature of 22-26°C and an air humidity of 45-65%. The animals were allowed to eat and drink freely and were adaptively fed for two weeks.
[0042] 1.2 Reagents
[0043] Total saponins from stems and leaves of American ginseng (purity 99.0%) were purchased from Ji'an Yisheng Pharmaceutical Co., Ltd., Jilin Province;
[0044] Estradiol valerate tablets were purchased from Guangzhou Bayer Healthcare Company;
[0045] Pentobarbital sodium was purchased from Sinopharm Chemical Reagent Co., Ltd.;
[0046] ELISA kits were purchased from Nanjing Jiancheng Biological Company Research Institute.
[0047] 1.3 Establishment of postmenopausal osteoporosis model
[0048] After two weeks of adaptive feeding of mice, the mice were weighed one day in advance, and 1% sodium pentobarbital injection was injected from the lower abdomen at 25g / 0.1mL; after successful anesthesia, the mice were placed in a prone position and fixed on the operating table, the back hair was removed, iodine was disinfected, and surgical scissors were used to cut the mouse along the middle of the back between the two hind legs, about 1cm long; then the subcutaneous tissue was cut along the midline to the left and right sides, and the muscle layer was found and cut; the white fat tissue covering the ovary was found, and the hemostatic forceps were used to separate the fat tissue, and the blood vessels under the ovary and other tissues were clamped and ligated; the ovary was cut off with surgical scissors, and the ovary on the other side was cut off in the same way; the fat tissue was restored, and the wound was rinsed with saline, and the wound was sutured, and the mouse was kept warm; it was put back in a clean cage, and the wound was cleaned with iodine for 1 week after surgery to prevent infection. After the operation was successful, the mouse recovered for 2 weeks, and the serum estrogen level of the mouse was detected by ELISA after blood was taken from the tail vein. According to the experimental results, the estrogen level was significantly reduced 2 weeks after ovariectomy, indicating that the mouse castration model was successful. After successful modeling, the mice were fed a high-fat diet for 14 weeks to construct a postmenopausal osteoporosis model.
[0049] 1.4 Experimental groups and drug administration method
[0050] 75 ApoE - / - Female mice and 10 C57BL / 6J female mice, after 2 weeks of adaptive feeding, C57BL / 6J female mice served as the control group; ApoE - / -Female mice were divided into the following five groups, 15 in each group: high-fat group (HFD), castration group (OVX), high-fat + castration group (HFD + OVX), high-fat + castration + PQS group (HFD + OVX + PQS), high-fat + castration + E2 group (HFD + OVX + E2) (see Table 1); according to the equivalent dose conversion coefficient table of animal and human body weight per kg, mice in each group were gavaged at 0.7 mL / 100 g. The control group, high-fat group, castration group, and high-fat + castration group were gavaged with the same amount of pure water; the high-fat + castration + PQS group was gavaged with 45 mg / kg of total saponins from American ginseng stems and leaves, once a day for 14 consecutive weeks; the high-fat + castration + E2 group was gavaged with 0.13 mg / kg of estradiol valerate tablets, once a day for 14 consecutive weeks. Serum and corresponding tissues were collected 24 hours after the last administration for detection.
[0051] Table 1
[0052] Serial number Group Dosage 1 Normal group Give an equal amount of pure water by gavage 2 High fat group Give an equal amount of pure water by gavage 3 Castration group Give an equal amount of pure water by gavage 4 High fat + castration group Give an equal amount of pure water by gavage 5 High fat + castration + PQS group Administer 45 mg / kg / day of total saponin solution from stems and leaves of American ginseng by intragastric administration 6 <![CDATA[High-fat + Ovariectomized + E2 group]]> Administer estradiol valerate aqueous solution 0.13 mg / kg / day by oral gavage
[0053] 2. Effect of total saponins from American ginseng stems and leaves on body weight of osteoporotic mice induced by high-fat diet and castration
[0054] During the 14-week experiment, the mice were weighed and recorded every 7 days, and the dosage was adjusted according to the weight. 24 hours after the last administration, the mice were weighed, anesthetized with intraperitoneal injection of sodium pentobarbital 40 mg / kg, and blood and corresponding tissues were taken for testing. Figure 1 ) showed that total saponins from American ginseng stems and leaves had no significant effect on the body weight of postmenopausal mice.
[0055] 3. Effects of total saponins from American ginseng stems and leaves on bone microstructure and bone mass in postmenopausal mice
[0056] Skyscan1276 X-Ray Micro CT was used to scan the entire femur. The bottom of the growth plate on the knee side of the femur was set as the scanning reference line. 83 consecutive slices were taken, that is, the area with a thickness of about 0.8 mm was set as the three-dimensional reconstruction region of interest (ROI); the bone volume fraction BV / TV (%), the number of trabeculae Tb.N (1 / mm), and the trabecular thickness Tb.Th (mm) in the bone tissue were analyzed; and the bone density BMD (g / cm 3 ). The experimental results showed that the number of trabeculae in the distal femur of the mice in the control group was large, the gaps were small, and the bone tissue was relatively dense; compared with the control group, the number of trabeculae in the model group was reduced, the gaps were enlarged, and the bone tissue was sparse; compared with the model group, after intervention with total saponins from American ginseng stems and leaves, the number of trabeculae increased, the gaps increased, and the bone tissue density increased. Experimental results ( Figure 2 ) indicates that total saponins from American ginseng stems and leaves can effectively counteract the decline in bone mass and bone volume in postmenopausal mice.
[0057] 4. Effects of total saponins from American ginseng stems and leaves on the expression of PINP and CTX-1 in the serum of postmenopausal mice
[0058] After the mice were anesthetized by intraperitoneal injection of 40 mg / kg sodium pentobarbital, non-anticoagulated blood was collected from the eye sockets and placed at -4°C for 2 hours. The blood was centrifuged at 3000 rpm for 20 minutes to separate the serum and stored at -80°C. The ELISA kit was used to detect the serum levels of PINP (ELISA kit) and CTX-1 (ELISA kit). Experimental results ( Figure 3 ) showed that total saponins from American ginseng stems and leaves could counteract the decrease in serum osteogenesis-related PINP levels and the increase in osteoclast-related CTX-1 levels in postmenopausal mice. After PQS treatment of postmenopausal mice, PINP levels increased and CTX-1 decreased.
[0059] 5. Effects of Total Saponins from American Ginseng Stems and Leaves on Histopathological Findings in Postmenopausal Mice
[0060] Take the femur and remove the skin and muscle; place the processed bone tissue in a 4% tissue fixative in a 4°C refrigerator overnight, rinse with deionized water several times, and then decalcify with 14% ethylenediaminetetraacetic acid (EDTA) on a shaker for 1 month, and change the decalcification solution once a day; remove the decalcification solution, and repeatedly rinse with deionized water and then dehydrate; after dehydration, take out the specimen, place it in the liquid wax pool on the paraffin embedding machine, and embed the inner side of the femur toward the bottom sagittal face, and after the embedded specimen is cooled and solidified on a low-temperature ice table, place it in a -20°C refrigerator for slicing; after the wax block is trimmed, slice it, start from the inner side, the slice thickness is 5μm, and observe while cutting and unfolding the slice, collect the articular surface slices in the center of the joint, and place them in a 37°C constant temperature box to dry overnight. Before dewaxing and hydration, place the tissue slices in a 60°C electric constant temperature blast drying oven for 2h. Dewaxing and hydration were performed in xylene (I, II) and gradient ethanol (95%, 90%, 80%, 70%, 50%), each time for 5 min.
[0061] A. Conventional HE staining: expression of basic morphology of bone tissue and number of osteoblasts;
[0062] B. TRAP staining of femur: osteoclast morphology.
[0063] Experimental results ( Figure 4 and Figure 5 ) showed that total saponins from American ginseng stems and leaves could counteract the decline in osteoblast function and the increase in osteoclasts in postmenopausal mice.
[0064] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims
1. Application of total saponins from American ginseng stems and leaves in the preparation of drugs for the treatment of postmenopausal osteoporosis.
2. The use according to claim 1, characterized in that: The total saponins of American ginseng stems and leaves include ginsenoside Rd, ginsenoside Rb3, ginsenoside Rb2 and pseudo-ginsenoside F11.
3. The use according to claim 2, characterized in that: The total saponins of American ginseng stems and leaves also include ginsenoside Rc, ginsenoside Re, ginsenoside Rg1, ginsenoside Rg2, ginsenoside Rg3 and ginsenoside F2.
4. The use according to claim 1, characterized in that: The postmenopausal osteoporosis is caused by postmenopausal estrogen deficiency.
5. The use according to claim 1, characterized in that: The postmenopausal osteoporosis includes early postmenopausal osteoporosis and late postmenopausal osteoporosis.
6. The use according to claim 1, characterized in that: Symptoms of postmenopausal osteoporosis include hunchback, decreased stature, and bone pain.
7. The use according to claim 6, characterized in that: The symptoms of postmenopausal osteoporosis also include local tenderness and percussion pain.
8. The use according to claim 1, characterized in that: The complications of postmenopausal osteoporosis include fractures, chest tightness, shortness of breath, cough, abdominal distension and constipation.
9. The use according to claim 1, characterized in that: The medicine contains an effective amount of total saponins from the stems and leaves of American ginseng.
10. The use according to claim 9, characterized in that: The mass proportion of total saponins in the stems and leaves of American ginseng in the medicine is 16.7%.