Application of flax lignan and flaxseed polysaccharide in preparation of health care product for improving osteoporosis

Abstract

The application discloses application of a lignan and flaxseed polysaccharide in preparation of health care products for improving osteoporosis, and belongs to the technical field of health care functions. The postmenopausal osteoporosis model is established by ovariectomized rats, and the comparison of the lignan and flaxseed polysaccharide on bone metabolism, related bone conversion indexes, bone microstructure, bone biomechanics and intestinal flora of the rats is systematically studied, so that the dominant material for improving osteoporosis of the lignan and flaxseed polysaccharide is explored, and a more effective theoretical basis is provided for development of functional food of flaxseed.

Description

Technical Field

[0001] This invention relates to the field of health function research technology, and more specifically to the application of linolenic acid and linseed polysaccharide in the preparation of health products to improve osteoporosis. Background Technology

[0002] my country is a major global producer of flaxseed, with extremely abundant reserves. Currently, flaxseed meal is mainly used as feed or fertilizer for ruminants, with only a small amount applied to the food industry. A large amount of byproducts remain unutilized, resulting in severe resource waste and environmental pollution. Flaxseed meal is rich in protein, dietary fiber, flax lignans, and other bioactive components, making it a biomass resource with significant development potential.

[0003] Flaxseed meal contains 35%–45% dietary fiber, with flaxseed gum being the main water-soluble dietary fiber and a high-quality flaxseed polysaccharide. Physiologically, flaxseed polysaccharides can regulate glucose and lipid metabolism, inhibit blood sugar elevation, and lower serum triglyceride and cholesterol levels, exhibiting potential for antioxidant and anti-tumor applications.

[0004] Flax lignans are characteristic high-value-added active ingredients in flaxseed meal, mainly existing in the form of open-ring isolar resinol diglucoside (SDG). Their content is far higher than that of common crops such as grains, fruits, vegetables, and legumes, making them an important source of natural lignans. Flax lignans possess phytoestrogen activity, exerting anti-estrogenic effects and inhibiting tumor growth, thus having application value in the intervention of diseases such as breast cancer. Their antioxidant activity is superior to vitamin E, effectively scavenging free radicals and alleviating oxidative stress. Simultaneously, they can regulate blood lipids and blood sugar, improve atherosclerosis, and have significant health benefits in cardiovascular protection and metabolic regulation.

[0005] Currently, existing technologies are being researched on the preparation and single function of flaxseed polysaccharides and flax lignans, but there are no reports on their interaction, combined application and synergistic effect mechanism, nor are there any related product development and industrial applications.

[0006] Therefore, exploring the synergistic mechanism of flaxseed polysaccharides and flax lignans is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0007] In view of this, the present invention provides the application of flax lignans and flaxseed polysaccharides in the preparation of health products for improving osteoporosis.

[0008] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0009] The application of flax lignans and flaxseed polysaccharides in the preparation of health products to improve osteoporosis, with a mass ratio of flax lignans to flaxseed polysaccharides of (1~4):1.

[0010] Furthermore, the mass ratio of linolenic acid to linseed polysaccharide is 2:1.

[0011] Furthermore, the mass ratio of linolenic acid to linseed polysaccharide is 4:1.

[0012] Furthermore, the osteoporosis mentioned is postmenopausal osteoporosis.

[0013] Furthermore, it is used for: a) Inhibit abnormal weight gain caused by oophorectomy; b) Increase serum estrogen levels; c) Reduce serum levels of osteocalcin, bone alkaline phosphatase, and tartrate-resistant acid phosphatase 5b. d) Increase femoral bone mineral density, bone volume fraction, number of trabeculae and connection density, and reduce trabecular separation; e) Improve the biomechanical properties of the femur; f) Regulate gut microbiota.

[0014] Furthermore, the preparation method of the flaxseed polysaccharide is as follows: flaxseed meal is pulverized, defatted, water-extracted and alcohol-precipitated, and then freeze-dried.

[0015] Furthermore, the dosage form of the health product is powder, tablet, capsule or granule.

[0016] As can be seen from the above technical solution, compared with the prior art, the present invention has the following beneficial effects: This invention establishes a postmenopausal osteoporosis model in ovariectomized rats and systematically studies the effects of flaxseed polysaccharides and flax lignans on bone metabolism, related bone turnover indicators, bone microstructure, bone biomechanics, and gut microbiota in rats. The aim is to explore the main active substances of flaxseed polysaccharides and flax lignans on osteoporosis improvement, thereby providing a more effective theoretical basis for the development of flaxseed functional foods. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0018] Figure 1 This is a comparison of the uterus and fallopian tubes of rats in each group in Example 1 of the present invention.

[0019] Figure 2 This is a comparison of the microstructure of the trabecular bone of the femur in each group of rats in Example 1 of the present invention.

[0020] Figure 3 This is a petal diagram of the OTUs in Embodiment 1 of the present invention.

[0021] Figure 4 The relative abundance of gut microbiota at the phylum (A) and genus (B) levels and the F / B value (C) are shown in Example 1 of this invention.

[0022] Figure 5 For the comparison of Beta diversity analysis in Example 1 of the present invention, note: A: PCoA; B: NMDS. Detailed Implementation

[0023] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. 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 are within the scope of protection of the present invention.

[0024] Example 1 1. Materials and Equipment 1.1 Materials and Reagents Flaxseed polysaccharide (FG), laboratory extracted; flax lignans (SDG), Xi'an Weisbo Biotechnology Co., Ltd.; SPF-grade female SD rats, 7-8 weeks old, weighing 180-220 g, fed (estrogen-free), Beijing Sibefore Biotechnology Co., Ltd.; Bone alkaline phosphatase (BALP), tartrate-resistant acid phosphatase 5b (TRAP-5b), serum osteocalcin (BGP), and estrogen (E2) kits, Shanghai Enzyme-Linked Biotechnology Co., Ltd.

[0025] Flaxseed meal was pulverized and passed through an 80-mesh sieve. The pulverized flaxseed meal was mixed with n-hexane at a ratio of 1:15 and stirred for 24 hours before being filtered. Distilled water was added to the defatted flaxseed meal powder, and the mixture was heated in a constant temperature water bath at 80°C for 2 hours. The mixture was then filtered through a 300-mesh filter to obtain the filtrate. The filtrate was concentrated under reduced pressure at 60°C to 50% of its original volume to obtain the crude extract. Four times the volume of anhydrous ethanol was added, and the mixture was refrigerated at 4°C for 12 hours. After centrifugation at 3500 r / min for 15 minutes, the precipitate was redissolved in five times the volume of water. The ethanol was removed by rotary evaporation, and the extracted flaxseed polysaccharide (FG) was obtained by vacuum freeze-drying.

[0026] 1.2 Instruments and Equipment Quantum GX2 Micro CT, Platinum Elmore; E10000 Universal Material Testing Machine, SMS, UK; Spark Multifunctional Microplate Reader, Tecron (Shanghai) Trading Co., Ltd.

[0027] 2. Experimental Methods 2.1 Preparation of animal models Fifty-six SPF-grade female SD rats were randomly divided into a sham-operated group (n=7) and a surgical group (n=49) after one week of acclimatization. In the surgical group, rats were anesthetized with isoflurane, fixed on a temperature-controlled plate, and shaved. The skin surface was wiped with iodine-soaked cotton balls. A 1.5 cm incision was made along the midline of the back, severing the mucous membrane between the skin and muscles on both sides of the back. The muscles on both sides were then longitudinally incised with wounds approximately 1 cm long. The pink, honeycomb-like ovary, encased in fat, was removed with flat forceps. The fallopian tubes and surrounding blood vessels were ligated with absorbable sutures. The ovary was removed, and the fallopian tubes were gently returned to the abdominal cavity. 3-4 drops of antibiotics were instilled to prevent infection. The wounds were neatly sutured, and the skin periphery was wiped with iodine-soaked cotton balls. The skin wounds were then fastened with a suture device. In the sham-operated group, the fat surrounding the ovary was removed, and the fallopian tubes were not ligated. Rats were injected with 20,000 U gentamicin for three consecutive days, and their mental state was observed.

[0028] 2.2 Animal grouping One week after surgery, 49 ovariectomized rats were randomly divided into a model (OVX) group, a positive control (E2) group, a linolenic acid (SDG) group, a low-dose, medium-dose, and high-dose mixture of linolenic acid and linseed polysaccharide (S / F) group, and a linseed polysaccharide (FG) group. All rats were administered the mixture by gavage as follows: Sham group and OVX group: treated with equal volume of distilled water; Group E2: Estradiol 1.5 mg / kg administered orally (equivalent to 14 times the daily clinical adult dose); Low-dose group of flax lignans and flaxseed polysaccharide mixture (LS / F): flax lignans 50 mg / kg + flaxseed polysaccharide 50 mg / kg by gavage; Medium-dose group (MS / F) of flax lignans and flaxseed polysaccharide mixture: 100 mg / kg flax lignans + 50 mg / kg flaxseed polysaccharide by gavage; High-dose group of flax lignans and flaxseed polysaccharide mixture (HS / F): flax lignans 200 mg / kg + flaxseed polysaccharide 50 mg / kg by gavage; SDG group: 100 mg / kg of linolenic acid was administered by gavage; FG group: 50 mg / kg of flaxseed polysaccharide was administered by gavage.

[0029] Each group was administered the solution via gavage once daily for 8 consecutive weeks, with a gavage volume of 1 mL / 100 g. The rats were fed a normal diet without estrogen and provided with ample water. Their body weight was measured twice a week.

[0030] 2.3 Collection and testing of various specimens After 8 weeks of continuous gavage, blood was collected from the heart under isoflurane anesthesia, centrifuged at 3000 r / min for 15 min, serum was separated, tubed and stored at -80℃ to measure bone turnover indicators related to bone metabolism.

[0031] The uterus, fallopian tubes, liver, and kidneys were collected, surrounding fat was removed, and uterine index and organ index were calculated.

[0032] The right femur of a rat was harvested, cleaned, and wrapped in gauze soaked in physiological saline for femoral biomechanical testing. The trabeculae were observed, and relevant metabolic indicators were measured.

[0033] Under aseptic conditions, approximately 0.2 g of fecal tissue was extracted from the colon of rats, placed in a 1.5 mL EP tube, labeled with the appropriate group, and frozen at -80 ℃ for later use.

[0034] 2.4 Measurement of Uterine Index and Organ Index The uterus was removed, the attached adipose tissue was removed, the integrity and contractility of the uterus were observed and weighed, the intact kidneys and liver of the rat were removed, the bloodstains were washed away with physiological saline, dried and weighed.

[0035] Uterine index (%) = uterine mass / rat weight; Kidney index (%) = total weight of both kidneys / total weight of rat; Liver index (%) = liver mass / rat weight.

[0036] 2.5 Detection of serum bone turnover markers related to bone metabolism The serum was removed and thawed at room temperature. The levels of bone alkaline phosphatase (BALP), tartrate-resistant acid phosphatase 5b (TRAP-5b), osteocalcin (BGP), and estrogen (E2) in the serum were measured according to the ELISA kit procedure.

[0037] 2.6 Detection of Bone Trabecular Metabolic Indicators Mirco CT was used to observe trabeculae, and the relevant parameters included bone mineral density (BMD), bone volume / tissue volume (BV / TV), number of trabeculae (Tb.N), trabecular separation (Tb.Sp), and connectivity density (Conn.D).

[0038] 2.7 Femoral biomechanical measurements The maximum load, maximum deformation, elastic modulus, and bending stress of the femur were measured.

[0039] 2.8 Detection of rat gut microbiota The data were submitted to Beijing Novogene Biotechnology Co., Ltd., and gut microbiota analysis was performed using 16S rRNA gene sequencing technology.

[0040] 2.9 Data Statistics and Analysis Experimental results are expressed as mean ± standard deviation. Data were processed using Origin and Graphpad Prism 8.0 software. One-way ANOVA was used for comparisons among multiple groups. P <0.05 is statistically significant.

[0041] 3 Results and Discussion 3.1 Effects on body weight Table 1. Changes in body weight of rats in each group

[0042] Note: When comparing the same indicator with Sham, ( P <0.05), ( P <0.01); compared with OVX, #( P <0.05), ## ( P <0.01).

[0043] All rats in each group were raised, modeled, and administered medication in a clean environment and at a suitable temperature, and maintained normal physiological function throughout. As shown in Table 1, compared with Sham, the OVX group showed a significant increase in body weight. This is because successful ovariectomy led to fat accumulation in the abdomen, resulting in rapid weight gain. However, treatment with the SDG group, MS / F, HS / F group, and E2 group slowed down the abnormal weight gain in rats. FG and LS / F had no significant effect on body weight, indicating that medium and high doses of S / F, SDG, and estradiol reversed the changes in body weight gain caused by ovariectomy to some extent.

[0044] 3.2 Effects on the uterus and fallopian tubes of rats like Figure 1 Table 2 shows that the size of the uterus, the thickness and weight of the fallopian tubes indicate that ovariectomy cannot maintain the normal size of the uterus in adult rats, and the fallopian tubes are severely atrophied. Compared with the Sham group, the OVX group rats had significantly thinner fallopian tubes, significantly smaller uteruses, significantly lighter weights, and a highly significantly reduced uterine index. P <0.01).

[0045] 3.3 Effects on organ indices in rats The liver and kidneys are important organs, and the organ index refers to the ratio of the weight of each organ to the body weight. As shown in Table 2, compared with the OVX group, the liver index of rats in the E2 group was significantly increased. P <0.01), in addition, the liver index of rats in the OVX group and the experimental drug administration group was significantly lower than that in the Sham group. Compared with the Sham group, the kidney index of rats in the FG group was significantly reduced ( P <0.05, the kidney index of rats in the OVX group and other experimental drug administration groups was significantly reduced ( P <0.01); Compared with the OVX group, the liver index in the E2 group was significantly increased, while there were no significant differences in the organ indices among the other groups. However, observation of the liver and kidneys during dissection revealed no abnormalities in their organ morphology, indicating that neither SDG nor FG had any adverse effects on the rats.

[0046] Table 2. Changes in uterine index, liver index, and kidney index in rats of each group.

[0047] Note: When comparing the same indicator with Sham, ( P <0.05), ( P <0.01); compared with OVX, #( P <0.05), ## ( P <0.01).

[0048] 3.4 Comparison of bone turnover markers related to serum bone metabolism Table 3 shows the results of bone turnover indicators related to bone metabolism in rat serum after 8 weeks of drug administration.

[0049] Table 3. Serum levels of E2, BGP, BALP, and TRAP-5b in each group of rats.

[0050] Note: When comparing the same indicator with Sham, ( P <0.05), ( P <0.01); compared with OVX, #( P <0.05), ## ( P <0.01).

[0051] The comparison of estrogen (E2) levels in the serum of rats in each group is shown in Table 3. The estrogen level in the serum of the Sham group was significantly higher than that in the other groups. P <0.01), while the E2 content in the serum of rats in the OVX group was the lowest, and the E2 content in the serum of rats in the OVX group was 75.29% lower than that in the Sham group; compared with the OVX group, the estrogen content in the serum of rats in the FG group did not change significantly, and the difference was not statistically significant. P >0.05), the serum estrogen levels in the other 5 experimental drug-treated rat groups were all higher than those in the OVX group, with the HS / F group showing an increase of 166.90%, which was statistically significant. P <0.01).

[0052] Osteocalcin (BGP), an active polypeptide secreted by osteoblasts, participates in osteoblast differentiation and bone resorption regulation, and is a specific indicator reflecting osteoblast function. The results of BGP content in the serum of rats in each group are shown in Table 3. Among all groups, the OVX group had the highest BGP content in serum, while the Sham group had the lowest, and the difference was statistically significant. P <0.01); Compared with the OVX group, the serum BGP levels in all six experimental drug-treated rat groups decreased, with the BGP levels in the serum of E2, MS / F, HS / F, and FG rats being significantly reduced ( P <0.05, P <0.01).

[0053] Bone alkaline phosphatase (BALP), secreted by osteoblasts, and tartrate-resistant acid phosphatase 5b (TRAP-5b), secreted by osteoclasts, are two essential enzymes for maintaining bone metabolic homeostasis in vivo. Changes in their levels reflect the activity of osteoblasts and osteoclasts. When osteoporosis occurs, the dynamic transition between bone resorption and bone formation accelerates, leading to high bone turnover and abnormally elevated levels of BALP and TRAP-5b. Table 3 shows the serum BALP and TRAP-5b levels in rats. Compared to Sham, the OVX group showed a significant increase in serum BALP and TRAP-5b levels. P <0.01), SDG, FG and estradiol can alleviate the abnormal changes in BALP and TRAP-5b caused by osteoporosis. Compared with the OVX group, the serum levels of BALP and TRAP-5b in the HS / F group were significantly reduced. P (<0.01), although the FG group showed a decrease, the difference was not statistically significant.

[0054] The SDG and HS / F groups showed significant therapeutic effects, particularly in inhibiting the synthesis of BGP and BALP and suppressing TRAP-5b levels. This may be achieved through the following combined mechanisms: Firstly, overactive osteoblasts may lead to abnormal bone formation, resulting in porous and irregular bone tissue that is detrimental to bone structural stability. Therefore, inhibiting excessive osteoblast activity and reducing the secretion of markers such as BGP and BALP can help increase bone mineral density. Secondly, inhibiting osteoclast activity reduces bone resorption and further improves bone mineral density, thereby improving osteoporosis.

[0055] 3.5 Effects on trabecular bone microstructure and detection of trabecular bone-related metabolic indicators By observing bone microstructure, the state of osteoporosis in mice can be more intuitively reflected. Micro-CT, or high-resolution micro-CT, is a 3D imaging technology based on computed tomography. Using this scanning imaging system to scan the femur of mice can more intuitively reflect the condition of osteoporosis. The scan results are as follows: Figure 2 As shown, compared to the Sham group, the OVX group had a significantly reduced number of trabeculae, with sparse fractures, larger gaps, discontinuous fractures, and severe disruption of the bone microenvironment. Compared to the OVX group, the E2, SDG, LS / F, MS / F, HS / F, and FG groups showed partial recovery in the number, morphological distribution, and connectivity of trabeculae.

[0056] The results of the trabecular bone-related metabolic indicators are shown in Table 4.

[0057] Table 4 Comparison of bone trabecular metabolic indices among different groups of rats

[0058] Note: When comparing the same indicator with Sham, ( P <0.05), ( P <0.01); compared with OVX, #( P <0.05), ## ( P <0.01).

[0059] When bone resorption increases, Tb.Sp values ​​rise, while a decrease in Tb.Sp values ​​indicates an increase in the diameter and area of ​​trabeculae, reflecting the average width of the medullary canal between trabeculae. In osteoporosis, Tb.N decreases, and Tb.Sp values ​​increase. As shown in Table 4, compared to the Sham group, the OVX group showed a highly significant decrease in BMD, BV / TV, Tb.N, and Conn.Dn. P <0.01), Tb.Sp increased significantly ( PThe value <0.01 indicates that ovariectomized osteoporotic rats exhibited osteoporotic manifestations such as reduced trabecular bone number and thickness, and increased trabecular separation. Compared with the OVX group, the BMD, BV / TV, Tb.N, and Conn.Dn values ​​of the E2, SDG, and HS / F groups were all increased, while the Tb.Sp value was decreased, all showing significant differences compared with the OVX group, indicating that they have the effect of improving the spatial structure of trabecular bone in rats.

[0060] 3.6 Effects on femoral biomechanical properties The results of the effects on the biomechanical properties of the femur are shown in Table 5.

[0061] Table 5 Comparison of biomechanical properties of rat femurs in different groups

[0062] Note: When comparing the same indicator with Sham, ( P <0.05), ( P <0.01); compared with OVX, #( P <0.05), ## ( P <0.01).

[0063] As shown in Table 5, the maximum load, maximum deformation, elastic modulus, and bending stress of the femur in the OVX group were significantly lower than those in the Sham group. P <0.01); The biomechanical parameters of the femur in the drug-treated rats were improved compared with those in the OVX group. The maximum deformation in the MS / F group was significantly higher than that in the OVX group. The elastic modulus of the low- and high-dose S / F groups was significantly higher than that in the OVX group. The bending stress in the experimental drug-treated rats was significantly higher than that in the OVX group, and the HS / F group was extremely significantly higher than that in the OVX group. P <0.01).

[0064] 3.7 Effects on gut microbiota 3.7.1 Analysis of the composition of OTUs in the overall sample microbial diversity All samples were clustered into OTUs (Operational Taxonomic Units), and species annotations were performed, resulting in a petal diagram. The results are as follows: Figure 3As shown: The Sham, OVX, E2, SDG, LS / F, MS / F, HS / F, and FG groups shared 633 OTUs; the Sham group had 327 unique OTUs; the OVX group had 253 unique OTUs; the SDG group had 364 unique OTUs; the LS / F group had 278 unique OTUs; the MS / F group had 274 unique OTUs; the HS / F group had 335 unique OTUs; and the FG group had 324 unique OTUs. This indicates significant differences in the species composition of the gut microbiota among the groups. Ovarian removal led to changes in the composition of the rat gut microbiota, and supplementation with SDG and FG could activate certain gut bacteria to become dominant.

[0065] 3.7.2 Effects on the relative abundance of gut microbiota species like Figure 4 A, Figure 4 C. Based on the species annotation results, the top 10 species with the highest abundance at the phylum level in each group were selected, and a relative abundance bar chart of species was generated. The dominant phyla in the rat gut were Firmicutes, Verrucomicrobiota, and Bacteroidota. Compared with the Sham group, the relative abundance of Firmicutes was increased in the OVX group, while the relative abundance of Verrucomicrobiota and Bacteroidota was decreased; compared with the OVX group, the relative abundance of Verrucomicrobiota was increased in all five experimental groups, while the relative abundance of Firmicutes was decreased. When the gut microbiota was dysbiotic, the abundance ratio of Firmicutes to Bacteroidetes (F / B) increased. The F / B value was 15.88 in the OVX group and 5.38 in the Sham group, with the OVX group being significantly higher than the Sham group. P <0.05), the experimental groups were lower than the OVX group, indicating that ovariectomy led to dysbiosis of the gut microbiota in rats. Supplementation with SDG and FG changed the taxonomic composition of the gut microbiota in OVX rats and improved the composition structure of the gut microbiota.

[0066] Distribution of gut microbiota diversity at the genus level is shown in the figure. Figure 4B. The two most abundant bacterial groups were Lactobacillus and Lachnospiraceae_NK4A136. Compared with the Sham group, the relative abundance of Lactobacillus was increased and the relative abundance of Lachnospiraceae_NK4A136 was decreased in the OVX group; compared with the OVX group, the relative abundance of Lactobacillus was decreased in all five experimental groups, while the relative abundance of Lachnospiraceae_NK4A136 was increased in the SDG, MS / F, and HS / F groups. Lactobacillus can promote increased calcium solubility in the intestinal lumen and enhance passive transport of calcium via the bypass pathway. Osteoporosis may lead to changes in the intestinal environment, such as increased inflammation and decreased calcium absorption. The increased abundance of Lactobacillus may be a compensatory response of the intestinal flora to maintain intestinal homeostasis. Lachnospiraceae_NK4A136 is an important producer of short-chain fatty acids (SCFAs). Osteoporosis may lead to changes in the gut environment, inhibiting the growth of this bacterial genus and thus reducing the production of SCFAs. SCFAs (especially butyrate) play an important role in maintaining intestinal barrier function, regulating the immune system, and promoting bone health. Their reduction may further exacerbate osteoporosis. It is evident that in ovariectomized rats, the gut microbiota is imbalanced, and supplementation with SDG and FG improves the gut microbiota dysbiosis in rats.

[0067] 3.7.3 Alpha Diversity Analysis The alpha diversity method was used to analyze the microbial community diversity, including species richness (i.e., the number of species) and species evenness (i.e., the uniformity of species distribution). We selected the Chao1, Simpson, and Shannon indices to assess the alpha diversity of the gut microbiota in each group of rats, and the results are shown in Table 6. Compared with the Sham group, the Chao1, Shannon, and Simpson indices in the OVX group all increased, although the statistical significance was not significant, it still suggests an increase in the number of species in the gut microbiota of osteoporotic rats, and an increasing trend in diversity and evenness of distribution. Compared with the OVX group, the Chao1, Shannon, and Simpson indices in the experimental group decreased slightly, although the statistical significance was not significant, it still suggests a decrease in microbiota richness after SDG and FG gavage.

[0068] Table 6 Comparison of α Diversity Analysis Indices

[0069] Note: When comparing the same indicator with Sham, ( P <0.05), ( P<0.01); compared with OVX, #( P <0.05), ## ( P <0.01).

[0070] 3.7.4 Comparison of multiple samples of gut microbiota The Beta Diversity method was used to compare and analyze the microbial community composition of different samples. The results of principal coordinate analysis (PCoA) and non-metric multi-dimensional scaling (NMDS) at the OTU level are as follows: Figure 5 As shown, there were differences among the five osteoporosis treatment groups in the Sham and OVX groups, and species differences existed among the groups; the group interval between the HS / F group and the Sham group was small, demonstrating that high doses of SDG and FG could intervene in postmenopausal osteoporosis rats and alter their gut microbiota.

[0071] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0072] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. The application of flax lignans and flaxseed polysaccharides in the preparation of health products for improving osteoporosis, characterized in that, The mass ratio of flax lignans to flaxseed polysaccharides is (1~4):

1.

2. The application as described in claim 1, characterized in that, The mass ratio of flax lignans to flaxseed polysaccharides is 2:

1.

3. The application as described in claim 1, characterized in that, The mass ratio of flax lignans to flaxseed polysaccharides is 4:

1.

4. The application as described in claim 1, characterized in that, The osteoporosis mentioned refers to postmenopausal osteoporosis.

5. The application as described in claim 1, characterized in that, Used for: a) Inhibit abnormal weight gain caused by oophorectomy; b) Increase serum estrogen levels; c) Reduce serum levels of osteocalcin, bone alkaline phosphatase, and tartrate-resistant acid phosphatase 5b. d) Increase femoral bone mineral density, bone volume fraction, number of trabeculae and connection density, and reduce trabecular separation; e) Improve the biomechanical properties of the femur; f) Regulate gut microbiota.

6. The application as described in claim 1, characterized in that, The preparation method of the flaxseed polysaccharide is as follows: flaxseed meal is crushed, defatted, water-extracted and alcohol-precipitated, and then freeze-dried.

7. The application as described in claim 1, characterized in that, The dosage form of the health product is powder, tablet, capsule or granule.

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