A traditional Chinese medicine composition for treating osteoporosis combined with abnormal lipid metabolism and application thereof

Abstract

The application belongs to the technical field of traditional Chinese medicines, and discloses a traditional Chinese medicine composition for treating osteoporosis combined with abnormal lipid metabolism and application, and the technical scheme is characterized in that the traditional Chinese medicine composition is composed of the following raw medicinal materials in parts by weight: rhizoma drynariae 9-15 parts, psoralea 7-13 parts, dog spine 7-13 parts, medlar 7-13 parts, oyster 7-13 parts, ginseng 3-9 parts, notoginseng 1-3 parts, and amomum 3-9 parts. The traditional Chinese medicine composition can not only improve bone density and bone tissue microstructure, but also regulate systemic lipid metabolism disorder, and significantly reduce TG, CHO and LDL-C levels. The dual function of "osteoporosis and lipid metabolism adjustment" is an important innovation point of the application, changes the limitation that traditional osteoporosis treatment only focuses on bone metabolism itself, and fills the treatment blank that the prior art cannot simultaneously intervene osteoporosis and abnormal lipid metabolism.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and more specifically, to a traditional Chinese medicine composition and its application for treating osteoporosis complicated with lipid metabolism abnormalities. Background Technology

[0002] Postmenopausal osteoporosis (PMOP) is a systemic bone metabolic disease caused by the decline in estrogen levels in women after menopause. It is characterized by decreased bone mass, reduced bone density, deterioration of bone microstructure, and increased bone fragility, and is a major cause of systemic bone pain, spinal deformities, and fragility fractures of the vertebrae and hip joints in middle-aged and elderly women. Bone metabolism is maintained in dynamic balance by osteoblast-mediated bone formation and osteoclast-mediated bone resorption. Decreased bone formation capacity and / or increased bone resorption are the core pathological mechanisms of PMOP. During bone formation, the osteogenic and adipogenic differentiation functions of bone marrow mesenchymal stem cells (BMSCs) are closely related to the number and function of osteoblasts, and are key upstream links affecting bone formation. When pathological changes occur in the local microenvironment of bone tissue, the osteogenic differentiation capacity of BMSCs weakens, and they differentiate into adipogenic cells instead, leading to a large accumulation of adipose tissue in the bone marrow cavity, reduced osteoblast sources, and inhibited bone formation, ultimately resulting in bone loss. Epidemiological studies and clinical practice have shown that osteoporosis patients often have concurrent lipid metabolism disorders, and there is a close pathophysiological link between the two. Postmenopausal estrogen deficiency not only leads to accelerated bone resorption but also causes lipid metabolism disorders, manifested as elevated levels of triglycerides (TG), total cholesterol (CHO), and low-density lipoprotein (LDL-C). This "bone-lipid comorbidity" phenomenon is particularly prominent in middle-aged and elderly populations, but currently, there is a lack of clinical treatment strategies that can simultaneously intervene in osteoporosis and lipid metabolism disorders.

[0003] Currently, clinical drugs for treating PMOP are mainly classified into three categories based on their mechanism of action: bone resorption inhibitors, bone formation promoters, and dual-action drugs. Bone resorption inhibitors are the most commonly used category in clinical practice, including bisphosphonates (such as alendronate sodium and zoledronic acid), selective estrogen receptor modulators (such as raloxifene), calcitonin, and RANKL monoclonal antibodies (denosumab). Bone formation promoters are mainly parathyroid hormone analogs (such as teriparatide). Dual-action drugs are represented by sclerosstatin monoclonal antibodies (romozolumab). These drugs mainly increase bone density and reduce fracture risk by inhibiting osteoclast function or promoting osteoblast activity. However, these drugs have significant side effects: long-term use of bisphosphonates can lead to serious adverse reactions such as osteonecrosis of the jaw and atypical femoral fractures; long-term use of selective estrogen receptor modulators and calcitonin carries a potential risk of tumor development, limiting their long-term clinical application. Furthermore, their therapeutic targets are singular: existing drugs mostly target only a single aspect of bone resorption or bone formation, lacking synergistic regulatory effects on bone metabolism and lipid metabolism. Commonly used bone resorption inhibitors in clinical practice mainly act on osteoclasts and have limited effects on promoting bone formation. Bone formation promoters such as teriparatide are limited by the duration of treatment (generally not exceeding 24 months), and bone resorption inhibitors need to be followed up after discontinuation to maintain bone mass. The regimen is cumbersome and patient compliance is poor.

[0004] Traditional Chinese medicine (TCM) possesses unique advantages in treating PMOP (porcine osteoporosis) due to its "multi-component, multi-target, and multi-pathway" approach and good safety profile. TCM theory posits that the pathogenesis of PMOP is a combination of deficiency and excess, with kidney deficiency as the root cause and blood stasis and phlegm turbidity as the manifestation. The kidneys govern bone and marrow production; abundant kidney essence ensures a sufficient source for bone marrow production, resulting in strong and powerful bones. Kidney deficiency leads to reduced marrow and bone wasting, manifesting as "bone atrophy." In TCM, "lipids" fall under the category of "phlegm turbidity." Kidney deficiency disrupts qi transformation and the abnormal distribution of body fluids, leading to fluid stagnation, phlegm production, and blood stasis. The mutual binding of phlegm and blood stasis obstructs the meridians, further exacerbating bone malnutrition. Therefore, osteoporosis and lipid metabolism disorders share a common origin in TCM theory, both closely related to kidney deficiency. This provides a theoretical basis for developing drugs that simultaneously improve bone and lipid metabolism. However, existing traditional Chinese medicine compound formulas and prepared Chinese medicines (such as Xianling Gubao Capsules, Gushukang Capsules, and Qianggu Capsules) mostly treat osteoporosis by tonifying the kidneys and strengthening bones, and are promoted and applied mainly by increasing bone density and improving bone microstructure. However, the specific signaling pathways by which they regulate bone metabolism are still unclear, and there is a general lack of systematic research on their regulatory effects on lipid metabolism, which cannot meet the comprehensive treatment needs of patients with osteoporosis and comorbid lipid metabolism disorders.

[0005] Therefore, the present invention provides a traditional Chinese medicine composition for treating osteoporosis complicated with lipid metabolism abnormalities and its application. Summary of the Invention

[0006] This invention provides a traditional Chinese medicine composition for treating osteoporosis complicated with lipid metabolism disorders and its application. The composition can promote bone formation, increase bone density, and regulate lipid metabolism disorders, aiming to solve the problem of the lack of drugs that can simultaneously intervene in bone metabolism and lipid metabolism in current clinical treatment.

[0007] Specifically, the present invention is achieved through the following technical solutions:

[0008] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating osteoporosis complicated with lipid metabolism abnormalities, the traditional Chinese medicine composition comprising the following raw medicinal materials in parts by weight: 9-15 parts of Drynaria fortunei, 7-13 parts of Psoralea corylifolia, 7-13 parts of Cibotium barometz, 7-13 parts of Lycium barbarum, 7-13 parts of Ostrea gigas, 3-9 parts of Panax ginseng, 1-3 parts of Panax notoginseng, and 3-9 parts of Amomum villosum.

[0009] Preferably, the traditional Chinese medicine composition consists of the following raw medicinal materials in parts by weight: 9 parts of Drynaria fortunei, 7 parts of Psoralea corylifolia, 7 parts of Cibotium barometz, 7 parts of Lycium barbarum, 7 parts of Ostrea gigas, 3 parts of Panax ginseng, 1 part of Panax notoginseng, and 3 parts of Amomum villosum.

[0010] In a second aspect, the present invention provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for treating osteoporosis complicated with lipid metabolism disorders.

[0011] Preferably, the preparation method of the drug includes: weighing the raw materials according to the weight parts of the above-mentioned traditional Chinese medicine composition, soaking them in water, decocting them at least twice, combining the filtrates from the two decoctions, and preparing any pharmaceutically acceptable dosage form with or without the addition of medically acceptable pharmaceutical excipients.

[0012] Preferably, the soaking time is 1 to 3 hours.

[0013] Preferably, the amount of water added during soaking is 6 to 10 times the total weight of the raw materials.

[0014] Preferably, the decoction includes: directly heating the soaked raw materials and medicinal liquid to boiling, decocting for 0.5 to 1.5 hours, filtering, adding 5 to 8 times the total weight of the raw materials to the dregs and decocting for at least 0.5 hours.

[0015] Preferably, the dosage form of the drug is an oral solid dosage form or an oral liquid dosage form, wherein the oral liquid dosage form is a decoction or syrup, and the oral solid dosage form is a powder, pill, granule, tablet or capsule.

[0016] Preferably, the pharmaceutical excipients include any one or more of fillers, disintegrants, lubricants, binders, preservatives, flavoring agents, and release regulators.

[0017] Preferably, the filler is any one or more of starch, lactose, sucrose, microcrystalline cellulose, mannitol, and inorganic salts; the disintegrant is any one or more of dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, and sodium croscarmellose; the lubricant is any one or more of talc, magnesium stearate, microcrystalline silica, polyethylene glycol, sodium dodecyl sulfate, and hydrogenated vegetable oil; and the binder is any one or more of starch paste, methylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and sodium carboxymethyl cellulose.

[0018] The formula analysis of the traditional Chinese medicine composition described in this invention is as follows:

[0019] Drynaria fortunei (Kunze) J. Sm., a plant in the Polypodiaceae family, is a dried rhizome. Its properties and channels are: bitter and warm. It enters the liver and kidney channels. Its functions and indications include: healing injuries and relieving pain, tonifying the kidneys and strengthening bones; externally, it dispels wind and removes blemishes. It is used for falls, sprains, fractures, kidney deficiency causing lower back pain, weakness of muscles and bones, tinnitus and deafness, and loose teeth; externally, it is used to treat alopecia areata and vitiligo.

[0020] Psoralea corylifolia L. is the dried, mature fruit of the legume Psoralea corylifolia L. Its properties and channels are: pungent, bitter, and warm. It enters the kidney and spleen channels. Its functions and indications include: warming the kidneys and tonifying yang, regulating qi and relieving asthma, warming the spleen and stopping diarrhea; externally, it is used to dispel wind and remove blemishes. It is used for kidney yang deficiency, impotence, seminal emission, enuresis, frequent urination, cold pain in the lower back and knees, asthma due to kidney deficiency, and diarrhea in the early morning; externally, it is used to treat vitiligo and alopecia areata.

[0021] Cibotium barometz (L.) J. Sm., a plant in the family Cibotaceae, is a dried rhizome. Its properties and channels are: bitter, sweet, and warm. It enters the liver and kidney channels. Its functions and indications are: dispelling wind and dampness, tonifying the liver and kidneys, and strengthening the lower back and knees. It is used for rheumatic pain, soreness and weakness of the lower back and knees, and weakness of the lower limbs.

[0022] Goji berries are the dried, ripe fruit of *Lycium barbarum* L., a plant in the Solanaceae family. Their properties and channels are: sweet and neutral. They enter the liver and kidney channels. Functions and indications: They nourish the liver and kidneys, benefit essence and improve eyesight. They are used for deficiency of essence and blood, soreness and weakness of the lower back and knees, dizziness and tinnitus, impotence and seminal emission, internal heat and thirst, blood deficiency and chlorosis, and blurred vision.

[0023] Oysters are the shells of the following oyster species: *Crassostrea gigas* (Thunberg), *Crassostrea talienwhanensis* (Crosse), or *Crassostrea ariakensis* (Wakiya). Properties and Channels Entered: Salty, slightly cold. Enters the Liver, Gallbladder, and Kidney channels. Functions and Indications: Calms the mind and spirit, nourishes Yin and subdues Yang, softens hardness and dissipates nodules. Used for palpitations, insomnia, dizziness, tinnitus, scrofula, phlegm nodules, and abdominal masses. Calcined oyster shell has astringent and consolidating properties, neutralizes acid and relieves pain. Used for spontaneous sweating, night sweats, seminal emission, metrorrhagia, leukorrhea, stomach pain, and acid regurgitation.

[0024] Ginseng is the dried root and rhizome of Panax ginseng CA Mey., a plant belonging to the Araliaceae family. Its properties and channels: sweet, slightly bitter, slightly warm. It enters the spleen, lung, heart, and kidney channels. Its functions and indications: greatly replenishes vital energy, restores the pulse and consolidates the body, tonifies the spleen and lungs, generates fluids and nourishes blood, calms the mind and improves intelligence. It is used for weakness and collapse, cold limbs and weak pulse, spleen deficiency and poor appetite, lung deficiency and cough, thirst due to fluid depletion, internal heat and thirst, deficiency of qi and blood, chronic illness and emaciation, palpitations and insomnia, impotence and cold uterus.

[0025] Panax notoginseng is the dried root and rhizome of Panax notoginseng (Burk.) FH Chen, a plant belonging to the Araliaceae family. Its properties and channels are: sweet, slightly bitter, and warm. It enters the liver and stomach channels. Its functions and indications include: dispersing blood stasis and stopping bleeding, reducing swelling and relieving pain. It is used for hemoptysis, hematemesis, epistaxis, hematochezia, metrorrhagia, traumatic bleeding, chest and abdominal pain, and swelling and pain from falls.

[0026] Amomum villosum Lour., Amomum villosum Lour. var. xanthioides TL Wu et Senjen, or Amomum longiligulare TL Wu are all plants belonging to the ginger family (Zingiberaceae). Properties and Channels Entered: Pungent, warm. Enters the Spleen, Stomach, and Kidney channels. Functions and Indications: Resolves dampness and invigorates the stomach, warms the spleen and stops diarrhea, regulates qi and calms the fetus. Used for dampness obstructing the middle jiao, epigastric fullness and loss of appetite, spleen and stomach deficiency-cold, vomiting and diarrhea, morning sickness, and threatened abortion.

[0027] This invention uses *Drynaria fortunei* as the principal herb, with *Psoralea corylifolia*, *Cibotium barometz*, *Lycium barometz*, and *Ostrea gigas* as assistant herbs, ginseng and *Panax notoginseng* as adjuvant herbs, and *Amomum villosum* as the guiding herb. *Drynaria fortunei* is bitter and warm, entering the liver and kidney meridians, and has the effects of tonifying the kidneys and strengthening bones, healing injuries and relieving pain; it is often used for kidney deficiency and lower back pain. *Psoralea corylifolia* enters the kidney and spleen meridians, and has the effects of warming the kidneys and assisting yang, promoting qi circulation, and warming the spleen and stopping diarrhea. *Cibotium barometz* enters the liver and kidney meridians, and has the effects of tonifying the liver and kidneys, strengthening the lower back and spine, and dispelling wind and dampness. *Lycium barometz* enters the liver and kidney meridians, and has the effects of nourishing the liver and kidneys. *Ostrea gigas* enters the liver and kidney meridians. These four assistant herbs are often used for symptoms such as lower back and knee weakness, lower limb weakness, and rheumatic pain. Ginseng enters the spleen, lung, heart, and kidney meridians, and has the effects of greatly tonifying vital energy and tonifying the spleen and lungs. *Panax notoginseng* enters the liver and stomach meridians, and has the effects of dispersing blood stasis, stopping bleeding, reducing swelling and relieving pain. *Amomum villosum* enters the spleen, stomach, and kidney meridians, and has the effects of resolving dampness, stimulating appetite, warming the spleen and stopping diarrhea.

[0028] The existing technology faces the following technical challenges in simultaneously intervening in bone metabolism and lipid metabolism: First, it is necessary to elucidate the dual regulatory mechanism of the formula on bone metabolism and lipid metabolism based on the clear efficacy of the formula; second, it is necessary to reveal the signaling pathways that exert the bidirectional regulatory effect at the molecular level; and third, the multi-component and multi-target characteristics of traditional Chinese medicine compound formulas have both advantages and bring complexity to mechanism research.

[0029] Compared with the prior art, the present invention has the following advantages:

[0030] 1. The dual therapeutic advantages of "simultaneous regulation of bone and lipids"

[0031] Existing osteoporosis treatments primarily target bone metabolism itself, while the traditional Chinese medicine composition provided by this invention not only increases bone density and improves bone tissue microstructure but also simultaneously regulates systemic lipid metabolism disorders, significantly reducing TG, CHO, and LDL-C levels. This dual effect of "bone and lipid regulation" is a key innovation of this invention, overcoming the limitation of traditional osteoporosis treatments that only focus on bone metabolism itself, and filling the gap in existing technologies that cannot simultaneously intervene in osteoporosis and lipid metabolism abnormalities.

[0032] 2. Clear multi-target molecular mechanism

[0033] This invention reveals for the first time the molecular mechanism by which the Bone-Nourishing and Marrow-Generating Formula regulates osteogenic / adipogenic differentiation of bone mesenchymal stem cells (BMSCs) by activating the PI3K / AKT / GSK-3β signaling pathway, and confirms the bidirectional regulatory effect of this formula on bone metabolism and lipid metabolism at the molecular level (promoting bone formation while inhibiting adipogenesis).

[0034] 3. Sound clinical foundation and safety

[0035] The safety test results of the bone-strengthening and marrow-generating formula of the present invention in animal experiments and the monitoring results of liver and kidney function showed no obvious adverse reactions, indicating that it is safe.

[0036] 4. The unique advantages of traditional Chinese medicine with multiple components and pathways

[0037] This invention embodies the unique advantages of traditional Chinese medicine (TCM) in its "multi-component, multi-target, and multi-pathway" approach, complementing the single-target intervention strategies of existing Western medicines. The kidney-tonifying herbs in the formula, such as Drynaria fortunei and Psoralea corylifolia, can activate bone marrow function and promote bone formation, while spleen-strengthening, qi-tonifying, and blood-activating herbs, such as ginseng and Panax notoginseng, can improve bone marrow microcirculation and immune function. The drug components work synergistically to regulate the body's internal environment and restore bone metabolic homeostasis. Attached Figure Description

[0038] Figure 1 Serum lipid metabolism levels in rats of each group (A: TG, B: CHO, C: LDL-C, D: HDL-C; n=5, compared with Ctrl) ### P<0.001, # P<0.05; compared with Sham △△△ P<0.001, △ P<0.05; compared with Mod *** P<0.001, ** P<0.01, * P<0.05, ns P>0.05);

[0039] Figure 2 Bone mineral density (BMD) and bone microstructure parameters of the femur in each group of rats are shown in Figure A, which is a 3D reconstruction image of the distal femur from micro-CT, with the first row being a longitudinal section and the second row being a transverse section; B, C, D, E, F, and G are BMD, BV / TV, Tb.N, Tb.Th, Tb.Sp, and SMI, respectively.

[0040] Figure 3 The results of ALP staining and Oil Red O staining in rats of each group and the statistical analysis are shown (A is a representative staining image, and B is the statistical analysis result of A).

[0041] Figure 4 The expression of PI3K, AKT, and GSK-3β proteins in rats of each group was detected by Western blot (A is the Western blot image, B, C, and D are the statistical analysis results of PI3K, AKT, and GSK-3β in A, β-actin is the internal reference protein, n=3).

[0042] Figure 5 Western blot was used to detect the expression of bone morphogenetic protein (RUNX2) and adipogenic differentiation protein (PPAR-γ) in rats of each group (A is the Western blot image, B and C are the statistical analysis results of RUNX2 and PPAR-γ in A, n=3).

[0043] Figure 6The levels of liver and kidney function biochemical indicators in each group of rats were (A represents AST, B represents ALT, C represents ALP, D represents CRE, n=5). Detailed Implementation

[0044] To enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be described in further detail below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0045] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other. The present invention will now be described in detail with reference to the embodiments.

[0046] Example 1

[0047] The traditional Chinese medicine composition provided in this embodiment for treating osteoporosis complicated with lipid metabolism abnormalities consists of the following raw medicinal materials in parts by weight: 9g of Drynaria fortunei, 7g of Psoralea corylifolia, 7g of Cibotium barometz, 7g of Lycium barbarum, 7g of Ostrea gigas, 3g of Panax ginseng, 1g of Panax notoginseng, and 3g of Amomum villosum.

[0048] Example 2

[0049] The traditional Chinese medicine composition provided in this embodiment for treating osteoporosis complicated with lipid metabolism abnormalities consists of the following raw medicinal materials in parts by weight: 15g of Drynaria fortunei, 13g of Psoralea corylifolia, 13g of Cibotium barometz, 13g of Lycium barbarum, 13g of Ostrea gigas, 9g of Panax ginseng, 5g of Panax notoginseng, and 9g of Amomum villosum.

[0050] Example 3

[0051] The traditional Chinese medicine composition provided in this embodiment for treating osteoporosis complicated with lipid metabolism abnormalities consists of the following raw medicinal materials in parts by weight: 12g of Drynaria fortunei, 10g of Psoralea corylifolia, 10g of Cibotium barometz, 10g of Lycium barbarum, 10g of Ostrea gigas, 6g of Panax ginseng, 3g of Panax notoginseng, and 6g of Amomum villosum.

[0052] Example 4

[0053] This embodiment provides a method for preparing a drug for treating osteoporosis complicated with lipid metabolism disorders, comprising the following steps:

[0054] According to the proportions of the traditional Chinese medicine composition in Examples 1, 2, or 3, weigh out Drynaria fortunei, Psoralea corylifolia, Cibotium barometz, Lycium barbarum, Ostrea gigas, Panax notoginseng, and Amomum villosum. Mix them together and add 6 to 10 times the total weight of the medicinal materials to purified water to soak for 1 to 3 hours. Heat to boiling and decoct for 0.5 to 1.5 hours. Filter to separate the liquid from the dregs. Add 5 to 8 times the amount of purified water to the dregs and continue to decoct for at least 0.5 hours after boiling. Combine the two filtrates, concentrate by water bath heating, and add or not add any pharmaceutically acceptable excipients to prepare any pharmaceutically acceptable dosage form.

[0055] Example 5

[0056] 1. Preparation of aqueous extract of the bone-strengthening and marrow-generating formula

[0057] Weigh out 12g of Drynaria fortunei, 10g of Psoralea corylifolia, 10g of Cibotium barometz, 10g of Lycium barbarum, 10g of Ostrea gigas, 6g of Panax ginseng, 3g of Panax notoginseng, and 6g of Amomum villosum. Mix them together and soak them in 8 times the total weight of the herbs in purified water for 2 hours. Heat the water to a boil and simmer at a constant temperature for 1 hour. Filter to separate the liquid from the dregs. Add 6 times the amount of purified water to the dregs and simmer at a constant temperature for 0.5 hours after the water boils. Combine the two filtrates and concentrate them in a water bath to a density of 1.4g / ml to obtain the aqueous extract of the Bone-Nourishing and Marrow-Generating Formula.

[0058] 2. Establishment and grouping of animal models

[0059] Eight-week-old SPF-grade female SD rats were used to establish a PMOP model using the OVX method. They were randomly divided into a model group (labeled Mod), a positive control group (labeled P-Ctrl), a low-dose Bu Gu Sheng Sui Fang group (labeled L-BGSSD), a medium-dose Bu Gu Sheng Sui Fang group (labeled M-BGSSD), and a high-dose Bu Gu Sheng Sui Fang group (labeled H-BGSSD). Healthy rats served as the blank control group (labeled Ctrl), and rats that underwent simulated surgery without actual intervention served as the sham surgery group (labeled Sham). Ten rats were in each group.

[0060] The positive control group was administered estradiol valerate at a dose of 0.088 mg·kg⁻¹·d⁻¹ via gavage. The low-dose, medium-dose, and high-dose groups of the Bu Gu Sheng Sui Fang (a traditional Chinese medicine formula) were administered the following doses via gavage: 2.94 g·kg⁻¹·d⁻¹, 5.87 g·kg⁻¹·d⁻¹, and 11.74 g·kg⁻¹·d⁻¹, respectively (i.e., 0.5 mL, 1 mL, and 2 mL of 1.4 g / mL aqueous extract to the low, medium, and high-dose groups, respectively). The blank control group, sham-operated group, and model group were administered an equal volume of physiological saline via gavage. The parameters were determined using the Meeh-Rubner formula (based on the conversion of the animal body surface area of ​​an adult (60 kg) to that of an experimental rat (300 g). Relevant indicators were measured after three months of continuous gavage administration.

[0061] 3. Indicator Testing

[0062] ELISA was used to detect four lipid parameters (triglycerides (TG), total cholesterol (CHO), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C)) and safety indicators (liver and kidney function, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and serum creatinine (CREA)). Micro-CT was used to detect bone mineral density (BMD) and bone microstructural parameters (bone volume fraction (BV / TV), trabecular bone number (Tb.N), trabecular bone thickness (Tb.Th), trabecular bone separation (Tb.Sp), and structural model index (SMI)) in rat femurs. HE staining and Masson staining were used to observe morphological changes in bone tissue. ALP staining and Oil Red O staining were used to observe local bone metabolism and lipid metabolism status in bone tissue. Western blotting was used to detect changes in the expression of pathway proteins (PI3K, AKT, GSK-3β), bone metabolism markers (RUNX2), and lipid metabolism markers (PPAR-γ).

[0063] 4. Experimental Results

[0064] 4.1 Formulas for nourishing bones and marrow can improve lipid metabolism

[0065] like Figure 1 As shown, compared with the blank control group and the sham-operated group, the serum TG, CHO and LDL-C levels of rats in the model group were significantly increased (P<0.05), while the TG, CHO and LDL-C levels of rats in the medium and high dose groups of Bugu Shengsui Fang were significantly decreased (P<0.05), while there was no significant difference in HDL-C (P>0.05), demonstrating a significant regulatory effect on lipid metabolism disorders.

[0066] 4.2 Formulas for nourishing bone and promoting marrow production can improve bone density and bone microstructure.

[0067] like Figure 2 As shown, compared with the blank control group and the sham-operated group, the femoral bone mineral density (BMD) of rats in the model group was significantly reduced (P<0.001); the BMD of the positive control group and the medium and high dose groups of Bu Gu Sheng Sui Fang were significantly increased (P<0.05). Bone microstructure analysis showed that BV / TV, Tb.N, and Tb.Th were significantly reduced in the model group, while Tb.Sp and SMI were significantly increased (P<0.01 to P<0.001); the high dose group of Bu Gu Sheng Sui Fang could increase BV / TV and Tb.N, and decrease Tb.Sp and SMI (P<0.05), while the medium dose group of Bu Gu Sheng Sui Fang could also effectively increase Tb.Th (P<0.05).

[0068] 4.3 The formula for replenishing bone marrow can bidirectionally regulate fat-bone metabolism in bone tissue.

[0069] like Figure 3As shown, ALP staining and Oil Red O staining results indicated that, compared with the blank control group and the sham-operated group, the proportion of ALP staining in the model group was significantly reduced and the proportion of Oil Red O staining was significantly increased (P<0.001); the positive control group and the medium and high dose groups of Bugu Shengsui Fang significantly increased the proportion of ALP staining and decreased the proportion of Oil Red O staining (P<0.01), directly demonstrating the bidirectional regulatory effect of Bugu Shengsui Fang on the osteogenic / adipogenic differentiation of local BMSCs in bone tissue.

[0070] 4.4 Regulation of pathway proteins by the Bone-Nourishing and Marrow-Generating Formula

[0071] like Figure 4 As shown, Western blotting results indicated that compared with the blank control group and the sham-operated group, the expression of PI3K and AKT in the model group rats was decreased, while the expression of GSK-3β was increased (P<0.001). The expression of PI3K and AKT was significantly increased and the expression of GSK-3β was significantly decreased in the medium and high dose groups of Bugu Shengsui Fang (P<0.05), indicating that Bugu Shengsui Fang exerts its anti-PMOP effect by regulating the PI3K / AKT / GSK-3β pathway.

[0072] 4.5 Regulation of osteogenic / adipogenic differentiation proteins by the bone-strengthening and marrow-generating formula

[0073] like Figure 5 As shown, Western blotting results indicated that compared with the sham-operated group, the expression of Runx2 in rats in the model group was decreased, while the expression of PPAR-γ was increased (P<0.001). The expression of Runx2 was significantly increased and the expression of PPAR-γ was significantly decreased in the low, medium, and high dose groups of Bu Gu Sheng Sui Fang (a traditional Chinese medicine formula) (P<0.001). Since Runx2 is the main transcription factor controlling osteoblast differentiation and PPAR-γ is a key regulatory protein driving adipocyte differentiation, this indicates that Bu Gu Sheng Sui Fang can enhance bone formation and inhibit bone marrow adipogenesis, drive the osteogenic-adipogenic differentiation balance of bone marrow mesenchymal stem cells and promote bone formation, thus restoring the bone metabolism imbalance in pathological states such as osteoporosis.

[0074] 4.6 Safety Indicators

[0075] like Figure 6 As shown, compared with the blank control group and the sham surgery group, the expression of ALT, AST, ALP and CREA in the model group and the low, medium and high dose groups of Bugu Shengsui Fang were not significantly different and were within the normal range, indicating that the application of Bugu Shengsui Fang is safe.

[0076] In summary, the present invention systematically evaluates the bone-protecting and lipid-regulating effects of the Bugushengfu Formula by establishing an ovariectomized (OVX) rat model, and clarifies the molecular mechanism by which it regulates the osteogenic / adipogenic differentiation of BMSCs through the PI3K / AKT / GSK-3β signaling pathway using methods such as serum pharmacology and pathway interference experiments. This provides a new solution to the current problems in the field of PMOP treatment, including insufficient promotion of bone formation, poor drug safety, and failure to meet the needs of comorbidity treatment, will reveal the scientific connection between the theory of "kidney governing bones and generating marrow" and modern bone metabolism research, and provide brand-new research and development ideas for kidney-tonifying traditional Chinese medicines to treat bone metabolic diseases.

[0077] This specific embodiment is merely an illustration of the present invention, not a limitation thereto. Those skilled in the art may make non-inventive modifications to this embodiment as needed after reading this specification, and all such modifications shall be protected by the patent law as long as they fall within the scope of the claims of the present invention.

Claims

1. A traditional Chinese medicine composition for treating osteoporosis complicated with lipid metabolism abnormalities, characterized in that, The traditional Chinese medicine composition consists of the following raw medicinal materials in parts by weight: 9-15 parts of Drynaria fortunei, 7-13 parts of Psoralea corylifolia, 7-13 parts of Cibotium barometz, 7-13 parts of Lycium barbarum, 7-13 parts of Ostrea gigas, 3-9 parts of Panax ginseng, 1-3 parts of Panax notoginseng, and 3-9 parts of Amomum villosum.

2. The traditional Chinese medicine composition for treating osteoporosis complicated with lipid metabolism abnormalities according to claim 1, and its application, characterized in that, The traditional Chinese medicine composition consists of the following raw medicinal materials in parts by weight: 9 parts of Drynaria fortunei, 7 parts of Psoralea corylifolia, 7 parts of Cibotium barometz, 7 parts of Lycium barbarum, 7 parts of Ostrea gigas, 3 parts of Panax ginseng, 1 part of Panax notoginseng, and 3 parts of Amomum villosum.

3. The use of the traditional Chinese medicine composition according to claim 1 or 2 in the preparation of a drug for treating osteoporosis complicated with lipid metabolism abnormalities.

4. The application according to claim 3, characterized in that, The method for preparing the drug includes: weighing the raw materials according to the weight parts of the traditional Chinese medicine composition described in claim 1 or claim 2, soaking them in water, decocting them at least twice, combining the filtrates from the two decoctions, and preparing any pharmaceutically acceptable dosage form with or without the addition of medically acceptable pharmaceutical excipients.

5. The method according to claim 4, characterized in that, The soaking time is 1 to 3 hours.

6. The method according to claim 4, characterized in that, The amount of water added for soaking is 6 to 10 times the total weight of the raw materials.

7. The method according to claim 4, characterized in that, The decoction process includes: directly heating the soaked raw materials and medicinal liquid to boiling, decocting for 0.5 to 1.5 hours, filtering, adding 5 to 8 times the total weight of the raw materials to the dregs and decocting for at least 0.5 hours.

8. The method according to claim 4, characterized in that, The dosage form of the drug is an oral solid dosage form or an oral liquid dosage form. The oral liquid dosage form is a decoction or syrup, and the oral solid dosage form is a powder, pill, granule, tablet or capsule.

9. The method according to claim 4, characterized in that, The pharmaceutical excipients include any one or more of fillers, disintegrants, lubricants, binders, preservatives, flavoring agents, and release regulators.

10. The method according to claim 9, characterized in that, The filler is any one or more of starch, lactose, sucrose, microcrystalline cellulose, mannitol, and inorganic salts; the disintegrant is any one or more of dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, and sodium croscarmellose; the lubricant is any one or more of talc, magnesium stearate, microcrystalline silica, polyethylene glycol, sodium dodecyl sulfate, and hydrogenated vegetable oil; and the binder is any one or more of starch paste, methylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and sodium carboxymethyl cellulose.

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