Thyroid function regulating preparation based on astragalus mongholicus fermentation extract and preparation method of thyroid function regulating preparation

Abstract

The invention relates to the technical field of traditional Chinese medicine preparations, and particularly discloses a thyroid function regulation and control preparation based on an astragalus membranaceus fermentation extract and a preparation method thereof, and the preparation is technically characterized in that the preparation takes an extract obtained after astragalus membranaceus is fermented by specific microorganisms (such as lactobacillus plantarum) as an active ingredient. Systematic iodine deficiency rat model experiments prove that the fermentation extract has obviously better effects in the aspects of improving thyroid functions and forms (such as adjusting FT3, FT4, TSH and TPO-Ab), enhancing body antioxidant ability (such as improving activity of SOD and CAT of hippocampus and liver) and improving learning and memory behaviors than a traditional astragalus extract. The invention provides an effective scheme for developing an innovative traditional Chinese medicine preparation for comprehensively preventing and treating the iodine deficiency thyroid disease and complications thereof.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and more specifically, to a thyroid function regulating preparation based on Astragalus membranaceus fermentation extract and its preparation method. Background Technology

[0002] The thyroid gland is an important endocrine organ in the human body, and iodine deficiency is a major cause of endemic goiter and hypothyroidism. Currently, the clinical treatment for iodine deficiency thyroid diseases mainly relies on oral levothyroxine sodium tablets for exogenous hormone replacement. Although this treatment method can correct thyroid hormone levels, it has the following significant drawbacks: First, it is a "replacement" rather than a "regulation" or "repair," and cannot fundamentally improve the function and morphology of the thyroid gland itself; second, it requires lifelong medication, resulting in poor patient compliance and the risk of dosage overload or underdosing; more importantly, simple hormone supplementation cannot effectively address the systemic pathological changes caused by iodine deficiency, such as compensatory hyperplasia and damage of thyroid follicular epithelial cells, increased levels of systemic oxidative stress, and the potential for impaired learning and memory abilities due to damage to brain regions such as the hippocampus.

[0003] Astragalus membranaceus is a commonly used qi-tonifying herb in traditional Chinese medicine. Modern pharmacological studies have shown that it possesses various activities, including immunomodulatory, antioxidant, and anti-inflammatory effects. Previous research has explored the potential of astragalus or its compound formulations in the adjuvant treatment of thyroid diseases. However, astragalus extracts prepared using traditional methods (such as water extraction and alcohol extraction) have limited bioavailability of their active ingredients (such as astragaloside A and polysaccharides), and their compositional profiles are relatively fixed. Furthermore, their ability to synergistically regulate the multi-system and multi-target damage involved in the complex pathological state of iodine deficiency is insufficient, and their therapeutic efficacy needs further improvement.

[0004] Traditional Chinese medicine fermentation technology is a modern processing method that utilizes microorganisms and their enzyme systems to biotransform medicinal materials. This technology can alter the original components of medicinal materials, potentially generating new active substances, increasing the content of effective components, or improving their bioavailability, thereby enhancing efficacy or reducing toxicity. Currently, although there is research on Astragalus fermentation, it mainly focuses on its regulation of immune or intestinal function. There are no reports on the application of Astragalus fermented extract specifically to iodine deficiency models, and the systematic and dynamic evaluation and comparison of its comprehensive effects with traditional extracts in improving thyroid function and morphology, enhancing systemic (especially central nervous system) antioxidant capacity, and improving related cognitive and behavioral disorders. Existing technologies lack an Astragalus preparation with multi-target optimized efficacy for iodine deficiency thyroid diseases based on clear comparative experimental evidence. Therefore, this application proposes a thyroid function regulating preparation based on Astragalus fermented extract and its preparation method to address the above-mentioned problems. Summary of the Invention

[0005] The purpose of this invention is to solve the technical problems mentioned in the background section and to provide a thyroid function regulating agent based on Astragalus fermentation extract and its preparation method.

[0006] The above-mentioned objective of the present invention is achieved as follows: One aspect of the present invention provides a thyroid function regulating preparation based on Astragalus fermentation extract, the active ingredient of which is Astragalus fermentation extract; the Astragalus fermentation extract is obtained by microbial fermentation of Astragalus extract.

[0007] Furthermore, the microbial fermentation uses probiotics, which are one of the genera Lactobacillus, Yeast, or Bacillus.

[0008] Furthermore, the regulatory agent is used to improve thyroid dysfunction caused by iodine deficiency.

[0009] Furthermore, the improvement of thyroid dysfunction includes: increasing serum levels of free triiodothyronine (FT3) and free thyroxine (FT4), and / or decreasing levels of thyroid-stimulating hormone (TSH), thyroglobulin antibody (TG-Ab), and anti-thyroid peroxidase antibody (TPO-Ab).

[0010] Furthermore, the regulatory agent is also used to enhance the antioxidant capacity of iodine-deficient individuals, specifically by increasing the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in the hippocampus and / or liver tissue, and / or reducing the content of malondialdehyde (MDA) and nitric oxide (NO).

[0011] Furthermore, the regulatory agent is also used to improve learning and memory dysfunction associated with iodine deficiency.

[0012] Furthermore, the efficacy of the regulatory agent was verified by the following experimental methods: using an iodine-deficient Wistar rat model, a normal control group, a low-iodine model group, different doses of Astragalus extract, and different doses of Astragalus fermented extract were set up for comparison. Thyroid function, tissue antioxidant indicators, and Morris water maze behavioral indicators were dynamically detected on days 30, 60, and 90 after administration.

[0013] Furthermore, its dosage form is oral liquid, tablet, capsule, granule or powder.

[0014] The present invention also provides a method for preparing the thyroid function regulating agent based on Astragalus fermentation extract, comprising the following steps: (1) Preparation of Astragalus extract; (2) Sterilize the Astragalus extract, inoculate it with fermentation bacteria to ferment it, and obtain Astragalus fermentation liquid; (3) Post-process the Astragalus fermentation broth to obtain Astragalus fermentation extract; (4) Prepare the desired dosage form by combining the fermented extract of Astragalus membranaceus with pharmaceutically acceptable excipients.

[0015] Furthermore, the fermentation strain mentioned in step (2) is Lactobacillus plantarum, and the fermentation conditions are: temperature 30-37℃, static culture for 5-8 days; the inoculation amount is 3%-8% of the volume of Astragalus extract.

[0016] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention is the first to demonstrate the targeted enhancement effect of fermentation process on the efficacy of Astragalus membranaceus. It is the first to conduct a head-to-head comparison between fermented Astragalus membranaceus extract and ordinary extract in a specific pathological model of iodine deficiency. It also demonstrates that fermented Astragalus membranaceus extract produces statistically significant synergistic effects over ordinary Astragalus membranaceus extract in multiple dimensions, including regulating thyroid hormones and autoantibodies, reducing proliferative lesions of thyroid tissue, enhancing the activity of key antioxidant enzymes in the hippocampus and liver, and improving spatial learning and memory abilities. This reveals that fermentation process is a key technical means to optimize the efficacy of Astragalus membranaceus for iodine deficiency thyroid diseases.

[0017] 2. The formulation provided by this invention is based on a "synergistic component group" produced by Astragalus fermentation, which can simultaneously act on the hypothalamus-pituitary-thyroid axis, peripheral and central oxidative stress systems, and hippocampus-dependent learning and memory pathways, achieving multi-target, holistic, and synergistic intervention on the iodine deficiency pathological network. This provides new research ideas and experimental evidence for understanding the treatment of complex diseases with traditional Chinese medicine compound formulas or fermentation products.

[0018] 3. Through dynamic analysis of multiple dose groups and multiple time points (days 30, 60, and 90), this invention not only proves the effectiveness of the fermentation extract, but also clarifies the dose range for achieving the best overall efficacy (e.g., medium and high doses in the model). This dynamic and gradual analysis method provides direct and quantitative data support for determining the optimal therapeutic dose and course of treatment of this regulatory preparation, making product development more scientific and targeted, and avoiding the blindness of traditional drug use.

[0019] 4. This invention develops a single innovative traditional Chinese medicine preparation that can simultaneously intervene in the core pathological aspects of thyroid diseases (functional and morphological damage) and their common complications (such as oxidative stress damage and cognitive impairment). It is expected to provide a new treatment option for clinical use that is different from simple hormone replacement and has multiple effects. It is of great significance for improving patients' quality of life and preventing and treating endemic thyroid diseases, and has significant social benefits. Attached Figure Description

[0020] Figure 1This is a comparison of core thyroid function indicators in the embodiments of the present invention (day 90 of the experiment); Figure 2 This is a comparison of the antioxidant index of hippocampal tissue in the embodiments of the present invention (day 90 of the experiment); Figure 3 These are the results of the Morris water maze behavioral test in this embodiment of the invention. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0022] The implementation of the present invention will be described in detail below with reference to specific embodiments.

[0023] This invention provides a thyroid function regulating agent based on Astragalus fermentation extract and its preparation method, aiming to provide an Astragalus preparation with multi-target optimized efficacy against iodine deficiency thyroid diseases based on clear comparative experimental evidence. This preparation is prepared through a specific microbial fermentation process and has been validated through systematic animal experiments, demonstrating superior comprehensive effects compared to traditional Astragalus extracts in improving thyroid function and morphological damage caused by iodine deficiency, enhancing the body's antioxidant capacity, and improving learning and memory impairment.

[0024] The active ingredient in this preparation is Astragalus fermentation extract, which is obtained by microbial fermentation of Astragalus extract. The preparation method is as follows: (1) Preparation of Astragalus extract.

[0025] (2) Sterilize the Astragalus extract, inoculate with fermentation bacteria to ferment and obtain Astragalus fermentation liquid; wherein, the fermentation bacteria is Lactobacillus plantarum, and the fermentation conditions are: temperature 30-37℃, static culture for 5-8 days; the inoculation amount is 3%-8% of the volume of Astragalus extract.

[0026] (3) Post-process the Astragalus fermentation broth to obtain Astragalus fermentation extract.

[0027] (4) Prepare the desired dosage form by combining the fermented extract of Astragalus membranaceus with pharmaceutically acceptable excipients.

[0028] The following is a specific implementation of the present invention: Example 1: Preparation of Astragalus Fermentation Extract and Ordinary Astragalus Extract 1. Preparation of Astragalus membranaceus extract (AE): Take 1 kg of Astragalus membranaceus slices, add 10 times the amount of pure water, and extract twice by reflux, 1.5 hours each time. Combine the extracts, filter, and concentrate the filtrate under reduced pressure at 60℃ to a thick paste. Take a portion of the thick paste and freeze-dry it to obtain a light yellow powder, which is the Astragalus membranaceus extract (AE), and store it in a sealed container.

[0029] 2. Preparation of Astragalus Fermentation Extract (FAE): The remaining Astragalus extract was adjusted to a crude drug concentration of 1 g / mL with pure water to obtain the Astragalus extract solution, which was sterilized at 121℃ for 20 minutes. Lactobacillus plantarum CGMCC 1.595 was inoculated into MRS liquid medium and activated for two generations. The activated bacterial solution was inoculated into the sterilized Astragalus extract solution at an inoculation rate of 5% (v / v), and fermented statically at 37℃ for 7 days. After fermentation, the fermentation broth was centrifuged at 4℃ and 8000 rpm for 15 minutes, and the supernatant was collected and filtered through a 0.22 μm microporous membrane for sterilization to obtain the Astragalus fermentation extract solution. This extract solution was spray-dried (inlet air temperature 180℃, outlet air temperature 85℃) to obtain a brownish-yellow powdered Astragalus fermentation extract (FAE), which was then sealed and stored.

[0030] Example 2: Pharmacodynamic Validation Experiment of the Modified Formulation 1. Experimental Animals and Grouping: 240 clean-grade 4-week-old Wistar rats, weighing (100±10) g, half male and half female, were randomly divided into 8 groups of 30 rats each after one week of acclimatization. Normal control group: fed with feed containing normal iodine content (approximately 300 μg I / kg) and free access to water.

[0031] Low-iodine model group: fed with low-iodine feed (iodine content <50 μg I / kg) and free access to water.

[0032] Low, medium, and high dose groups of Astragalus extract: fed with low-iodine diet, and administered 0.25, 0.50, and 1.00 mg·L⁻¹ of ordinary Astragalus extract (AE) aqueous solution by gavage daily.

[0033] Low, medium, and high dose groups of Astragalus fermented extract: fed with low-iodine diet, and administered 0.25, 0.50, and 1.00 mg·L⁻¹ of Astragalus fermented extract (FAE) aqueous solution by gavage daily.

[0034] All gavage volumes were 1 mL / 100g body weight. The normal control group and the model group were gavage with the same volume of pure water. The experimental period was 90 days.

[0035] 2. Dynamic sampling and detection: On days 30, 60 and 90 after the start of the experiment, 10 rats from each group were randomly sacrificed for index detection.

[0036] Behavioral testing (Morris water maze): The water maze experiment was conducted 6 days before each sacrifice. The rats were trained to navigate to the hidden platform for 4 consecutive days, and the escape latency (in seconds) was recorded. On the 5th day, the platform was removed and a spatial exploration experiment was conducted. The number of times the rats crossed the original platform position within 60 seconds was recorded to assess their spatial learning and memory abilities.

[0037] Sample collection: Weigh the animal before sacrifice, collect blood from the abdominal aorta after anesthesia, and separate the serum; quickly remove the thyroid gland, intact hippocampus, and part of the liver tissue.

[0038] Indicator Testing: a) Thyroid function and morphology: Serum FT3, FT4, TSH, TG-Ab, and TPO-Ab levels were measured using chemiluminescent immunoassay. Thyroid tissue was fixed in 10% neutral formalin, routinely embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE), and its morphology was observed and photographed under a light microscope.

[0039] b) Tissue antioxidant indicators: Accurately weigh hippocampal and liver tissues, add pre-cooled physiological saline to prepare a 10% tissue homogenate, centrifuge and collect the supernatant. Using the appropriate kit, strictly follow the instructions to determine the activities of CAT, SOD, and GSH-Px, as well as the contents of MDA and NO in the supernatant.

[0040] 3. Experimental results (e.g.) Figures 1-3 (as shown) Thyroid function and morphology: Serum FT3 and FT4 levels in the low-iodine model group were significantly lower than those in the normal control group (P<0.01), while TSH and TPO-Ab levels were significantly higher (P<0.01). Thyroid tissue showed typical pathological changes such as follicular epithelial cell hyperplasia and follicular lumen shrinkage. All treatment groups showed varying degrees of improvement. Compared with the same dose of AE, the FAE dose groups, especially the medium and high dose groups, showed statistically significant improvements in increasing FT3 and FT4, decreasing TSH and TPO-Ab, and alleviating thyroid tissue hyperplasia (P<0.05).

[0041] Antioxidant capacity: In the low-iodine model group, the activities of SOD, CAT, and GSH-Px in the hippocampus and liver tissues were significantly reduced, while the contents of MDA and NO were increased. After drug intervention, the medium- and high-dose FAE groups were significantly better than the AE group at the same dose in increasing the activities of SOD and CAT in the hippocampus and liver and reducing the contents of MDA (P<0.05).

[0042] Learning and memory behaviors: In the low-iodine model group, the escape latency in the water maze navigation test was significantly longer than that in the normal control group (P<0.01), and the number of platform crossings in the spatial exploration test was significantly reduced (P<0.01). In the medium- and high-dose FAE groups, the escape latency was significantly shorter than that in the same-dose AE group (P<0.05), and the number of platform crossings was significantly greater than that in the same-dose AE group (P<0.05), indicating that FAE is more effective in improving iodine deficiency-induced learning and memory impairment.

[0043] 4. Experimental Conclusion: This embodiment, through systematic analysis of multiple time points and multiple indicators, fully verifies that the Astragalus fermentation extract regulatory preparation described in this invention has comprehensive efficacy superior to traditional Astragalus extract in improving thyroid function and morphological damage caused by iodine deficiency, enhancing the body's antioxidant capacity, and improving cognitive dysfunction.

[0044] Example 3: Preparation of a Regulated Formulation (Tablet) Take 50g of Astragalus fermented extract (FAE) powder prepared in Example 1 (passed through an 80-mesh sieve), mix it evenly with 35g of microcrystalline cellulose, 10g of pregelatinized starch, 4g of sodium carboxymethyl starch and 1g of magnesium stearate, and compress it into tablets using the direct compression method to make tablets weighing about 0.5g each and containing 250mg of FAE.

[0045] Example 4: Preparation of a regulatory formulation (oral liquid) The Astragalus fermentation extract prepared in Example 1 was adjusted to a FAE concentration of 0.5 mg / mL with purified water. 5% (by volume) of honey was added as a flavoring agent and 0.05% (by volume) of ethylparaben as a preservative, and the mixture was stirred until completely dissolved. The solution was filtered through a 0.45 μm filter membrane, filled into 10 mL oral liquid bottles, sealed, and sterilized with flowing steam at 100°C for 30 minutes to obtain the oral liquid preparation.

[0046] Through the above embodiments of the present invention, this invention obtains an Astragalus fermented extract, which is completely different from traditional extracts, by fermenting Astragalus with specific microorganisms (such as Lactobacillus plantarum). Furthermore, for the first time, a multi-dimensional, dynamic comparative experimental system was constructed and applied to confirm that this fermented extract, in an iodine deficiency pathological model, synergistically and significantly outperforms traditional extracts, while simultaneously improving thyroid function and morphology, enhancing the body's (especially the hippocampus) antioxidant capacity, and reversing learning and memory impairments. This unexpected "multi-system synergistic effect" of the present invention provides a novel, empirically proven, and effective solution for addressing iodine deficiency diseases and their complications.

[0047] 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, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A thyroid function regulating agent based on Astragalus membranaceus fermentation extract, characterized in that, Its active ingredient is Astragalus fermentation extract; the Astragalus fermentation extract is obtained by microbial fermentation of Astragalus extract.

2. The thyroid function regulating agent based on Astragalus fermentation extract according to claim 1, characterized in that, The microbial fermentation uses probiotics, which are one of the following: lactic acid bacteria, yeast, or Bacillus.

3. The thyroid function regulating preparation based on Astragalus fermentation extract according to claim 1 or 2, characterized in that, The regulatory agent is used to improve thyroid dysfunction caused by iodine deficiency.

4. The thyroid function regulating agent based on Astragalus fermentation extract according to claim 3, characterized in that, The improvement of thyroid dysfunction includes: increasing serum levels of free triiodothyronine (FT3) and free thyroxine (FT4), and / or decreasing levels of thyroid-stimulating hormone (TSH), thyroglobulin antibody (TG-Ab), and anti-thyroid peroxidase antibody (TPO-Ab).

5. The thyroid function regulating preparation based on Astragalus fermentation extract according to claim 1 or 2, characterized in that, The regulatory agents are also used to enhance the antioxidant capacity of iodine-deficient individuals, specifically by increasing the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in the hippocampus and / or liver tissue, and / or reducing the levels of malondialdehyde (MDA) and nitric oxide (NO).

6. The thyroid function regulating preparation based on Astragalus fermentation extract according to claim 1 or 2, characterized in that, The regulatory agents are also used to improve learning and memory dysfunction associated with iodine deficiency.

7. The thyroid function regulating preparation based on Astragalus fermentation extract according to any one of claims 1-6, characterized in that, The efficacy of the regulatory agent was verified by the following experimental methods: using an iodine-deficient Wistar rat model, a normal control group, a low-iodine model group, different doses of Astragalus extract, and different doses of Astragalus fermented extract were set up for comparison. Thyroid function, tissue antioxidant indicators, and Morris water maze behavioral indicators were dynamically detected on days 30, 60, and 90 after administration.

8. The thyroid function regulating preparation based on Astragalus fermentation extract according to any one of claims 1-7, characterized in that, Its dosage forms include oral liquid, tablets, capsules, granules or powders.

9. A method for preparing the thyroid function regulating agent based on Astragalus membranaceus fermentation extract according to any one of claims 1-8, characterized in that, Includes the following steps: (1) Preparation of Astragalus extract; (2) Sterilize the Astragalus extract, inoculate it with fermentation bacteria to ferment it, and obtain Astragalus fermentation liquid; (3) Post-process the Astragalus fermentation broth to obtain Astragalus fermentation extract; (4) Prepare the desired dosage form by combining the fermented extract of Astragalus membranaceus with pharmaceutically acceptable excipients.

10. The method according to claim 9, characterized in that, The fermentation strain mentioned in step (2) is Lactobacillus plantarum, and the fermentation conditions are: temperature 30-37℃, static culture for 5-8 days; the inoculum amount is 3%-8% of the volume of Astragalus extract.

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