A health-care nutritional agent of cistanche tubulosa for improving intestinal flora and a preparation method thereof

Abstract

The present application relates to the technical field of health-care nutrients, in particular to a health-care nutrient of Cistanche tubulosa for improving intestinal flora and a preparation method thereof. The health-care nutrient is prepared from the following components in parts by weight: Cistanche tubulosa 20-30 parts, Imperata cylindra 4-6 parts, Radix Paeoniae Alba 2-4 parts, Portulaca oleracea 1-2 parts, Hericium erinaceus 2-4 parts, fructooligosaccharide 4-6 parts, resistant dextrin 2-3 parts, and steviol glycoside 0.1-0.2 parts. The obtained nutrient has strong antioxidant activity, can significantly promote the proliferation of probiotics such as Bifidobacterium, can effectively restore the weight loss caused by intestinal flora disorder, and can synergistically improve the intestinal microecology.

Description

Technical Field

[0001] This invention relates to the field of health supplement technology, specifically to a Cistanche tubulosa health supplement that improves intestinal flora and its preparation method. Background Technology

[0002] The intestines are the largest digestive organ in the human body, containing a large number of bacteria, archaea, eukaryotic microorganisms, and viruses, collectively known as the gut microbiota. The core function of the gut microbiota is to maintain homeostasis, making it a crucial part of the body's micro-ecological system. Changes in the quantity, structure, diversity, and abundance of the gut microbiota are closely related to the body's physiological and pathological states, especially its significant role in immunity. *Cistanche tubulosa*, also known as *Cistanche spp.*, primarily parasitizes the roots of plants in the *Tamarix* genus. Belonging to the *Cistanche* genus of the Orobanchaceae family, it is a traditional Chinese medicine for tonifying the middle energizer and replenishing qi. It is warm in nature, sweet and salty in taste, and enters the kidney and large intestine meridians. It has a wide range of medicinal functions, including tonifying kidney yang, nourishing essence and blood, and moistening the intestines to relieve constipation, earning it the reputation of "desert ginseng." Modern pharmacological studies have shown that *Cistanche tubulosa* has a wide range of pharmacological effects, primarily including tonifying the kidney and strengthening yang, moistening the intestines to relieve constipation, protecting the liver, anti-oxidation, anti-fatigue, anti-aging, and enhancing the body's immunity. Cistanche tubulosa is rich in active ingredients such as phenylethanoid glycosides and polysaccharides, which have effects such as regulating immunity, anti-oxidation, and promoting intestinal health. However, existing Cistanche tubulosa products are mostly used in the form of simple extracts, which have limited effects on regulating the intestinal microecology. Summary of the Invention

[0003] To address the shortcomings of existing technologies, this invention proposes a Cistanche tubulosa health supplement that improves intestinal flora and its preparation method.

[0004] This invention is achieved through the following technical solution: A health supplement containing Cistanche tubulosa to improve gut microbiota, the raw materials for which are prepared include the following components in parts by weight: 20-30 parts of Cistanche tubulosa, 4-6 parts of Imperata cylindrica root, 2-4 parts of Paeonia lactiflora, 1-2 parts of Portulaca oleracea, 2-4 parts of Hericium erinaceus, 4-6 parts of fructooligosaccharides, 2-3 parts of resistant dextrin, and 0.1-0.2 parts of steviol glycosides.

[0005] Furthermore, the Cistanche tubulosa health supplement for improving intestinal flora is prepared by including the following components in parts by weight: 0.05-0.08 parts of Lactobacillus paracasei, 0.03-0.05 parts of Lactobacillus reuteri, and 0.02-0.03 parts of Pediococcus pentosaceus, all of which are powder preparations with a live bacteria count of 1.5 × 10⁻⁶. 11 CFU / g.

[0006] Furthermore, the present invention also provides a method for preparing the aforementioned Cistanche tubulosa health supplement that improves intestinal flora, comprising the following steps: S1. Cistanche tubulosa, Imperata cylindrica root, Paeonia lactiflora and Portulaca oleracea are pulverized through an 80-100 mesh sieve to obtain Chinese herbal powder. The Chinese herbal powder is added to deionized water, soaked for 1 hour, boiled and then decocted for another 1 hour. The mixture is filtered to obtain residue and filtrate. S2. Sterilize the filtrate obtained in step S1 at 121℃ for 20 min, cool it, and then inoculate it with activated Lactobacillus paracasei, Lactobacillus reuteri and Pediococcus pentosus. Ferment at 37℃ for 48 h, centrifuge at 10000 rpm for 15-20 min, filter the supernatant through a 0.22 μm filter membrane, concentrate it under reduced pressure to 20% of the original volume, and freeze dry to obtain the fermentation extract. S3. Dry the filter residue obtained in step S1, add it to a 70-80 vol% ethanol solution, heat it under reflux in an 80°C water bath for 1 h, centrifuge it at 8000 rpm for 15-20 min, remove the ethanol from the supernatant by vacuum distillation, and freeze-dry it to obtain the filter residue extract. S4. Dry the Hericium erinaceus to constant weight, pulverize it through an 80-mesh sieve to obtain Hericium erinaceus powder, add it to a 0.2 mol / L phosphate buffer solution with pH 7.8, stir at 400-500 rpm for 4 h, centrifuge at 10000 r / min for 15-20 min, and take the supernatant. S5. Take the supernatant obtained in step S4, add ammonium sulfate to saturation, let stand overnight at 4°C, centrifuge, place the precipitate in a 3500 Da dialysis bag and dialyze in deionized water for 24 h, freeze-dry the dialysate to obtain Hericium erinaceus water-soluble protein; S6. The Hericium erinaceus water-soluble protein obtained in step S5 was added to PBS (0.01M) buffer at pH 6.8 and stirred overnight at 4°C. The pH was adjusted to 9 with 1 mol / L NaOH. The filter residue extract obtained in step S3 was added and stirred for 24 h. The mixture was dialyzed in deionized water for 48 h. The dialysate was freeze-dried to obtain the extract - Hericium erinaceus protein. S7. Take the extract obtained in step S6 - Hericium erinaceus protein, and the fermentation extract fermentation preparation obtained in step S2, fructooligosaccharides, resistant dextrin and steviol glycosides, and mix them evenly to obtain a Cistanche tubulosa health care nutritional agent that improves intestinal flora.

[0007] Furthermore, in step S1, the ratio of the traditional Chinese medicine powder to deionized water is 1 g: 20 mL.

[0008] Further, in step S3, the ratio of the filter residue to the ethanol solution is 1 g: 10-15 mL.

[0009] Further, in step S4, the ratio of the Hericium erinaceus powder to the phosphate buffer is 1 g: 20-30 mL.

[0010] Further, in step S6, the mass concentration of the Hericium erinaceus water-soluble protein in the PBS buffer is 20-25 mg / mL.

[0011] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a health supplement containing Cistanche tubulosa to improve gut microbiota. It extracts and releases phenylethanoid glycosides and polysaccharides from Cistanche tubulosa, as well as polyphenols, flavonoids, and organic acids from Imperata cylindrica root, Paeonia lactiflora, and Portulaca oleracea, among other antioxidant active substances. Hericium erinaceus water-soluble protein, under alkaline conditions, covalently or non-covalently couples with small-molecule phenolic active components in the alcohol extraction residue, forming an extract-Hericium erinaceus protein complex. This complex not only possesses a certain free radical scavenging ability but also protects unstable antioxidant components through intermolecular interactions, delaying their oxidative inactivation, thereby producing a synergistic antioxidant effect. This health supplement provides growth-promoting factors for probiotics. The fermented extract obtained from the aqueous extract of the traditional Chinese medicine, after being fermented by Lactobacillus paracasei, Lactobacillus reuteri, and Pediococcus pentosaceus, is rich in organic acids, short-chain fatty acids, peptides, and other post-biotic metabolites, which can act as signaling molecules or nutrient substrates to stimulate the growth of probiotics. Fructooligosaccharides and resistant dextrin, as prebiotics, can be fermented and utilized by Bifidobacteria and Lactobacillus, accelerating their proliferation. The complex formed by coupling Hericium erinaceus protein with the filter residue extract can achieve sustained release of small-molecule active substances in the digestive tract, maintaining an effective concentration in the colonic probiotic enrichment zone for a long time, providing stable support for the continuous proliferation of probiotics. The fermented extract of this invention can inhibit pathogenic bacteria, regulate intestinal pH, and enhance the expression of tight junction proteins in the intestinal epithelium, thereby rapidly repairing the damaged intestinal barrier; the protein-extract conjugate exerts sustained anti-inflammatory and antioxidant effects through the sustained release of polyphenols and other active molecules, reducing intestinal inflammation; fructooligosaccharides and resistant dextrin, among other prebiotics, promote the proliferation of endogenous beneficial bacteria, competitively exclude pathogenic bacteria, and restore the balance of the gut microbiota; the formula of multiple traditional Chinese medicines, including Cistanche tubulosa and Imperata cylindrica, synergistically tonifies the spleen and kidneys, moistens the intestines and promotes bowel movements, comprehensively improving intestinal digestive and absorptive functions. This invention uses the filtrate obtained from the water extraction of traditional Chinese medicine powder for probiotic fermentation, and further extracts active substances from the filter residue, achieving gradient utilization of raw materials. The extraction and purification conditions of Hericium erinaceus protein are mild, preserving the protein's natural structure and reactivity. Controllable coupling with small molecules extracted from alcohol significantly improves the water solubility and stability of active substances such as phenols, enhancing their retention time and bioavailability in vivo. Finally, the fermentation extract, protein conjugate, prebiotics, and natural sweetener steviol glycosides are mixed to obtain a product with synergistic functions, a pleasant taste, and multiple benefits including antioxidant activity, promotion of probiotic proliferation, and intestinal repair. This provides an efficient and feasible solution for the in-depth development and utilization of Cistanche tubulosa and for intervention in intestinal microecological health. Attached Figure Description

[0012] Figure 1The DPPH free radical scavenging rate of the health supplements described in Examples 1-3 and Comparative Examples 1-3 of this invention; Figure 2 The growth-promoting effects of the health supplements in Example 2 and Comparative Examples 1-3 of this invention on Bifidobacterium longum and Lactobacillus acidophilus are shown. Figure 3 The effects of the health supplements described in Example 1 and Comparative Examples 1-3 on the body weight of mice with gut microbiota dysbiosis. Detailed Implementation

[0013] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. However, this invention is not limited to the following embodiments. It should be noted that, unless otherwise specified, all chemical reagents involved in this invention are purchased through commercial channels.

[0014] Example 1: A health supplement containing Cistanche tubulosa to improve gut microbiota, the raw materials for preparation include the following components in parts by weight: 30 parts of Cistanche tubulosa, 6 parts of Imperata cylindrica root, 4 parts of Paeonia lactiflora, 2 parts of Portulaca oleracea, 4 parts of Hericium erinaceus, 6 parts of fructooligosaccharides, 3 parts of resistant dextrin, and 0.2 parts of steviol glycosides.

[0015] The Cistanche tubulosa health supplement for improving gut microbiota also includes the following components by weight: 0.08 parts of Lactobacillus paracasei, 0.05 parts of Lactobacillus reuteri, and 0.03 parts of Pediococcus pentosaceus, all in powder form, with a live bacteria count of 1.5 × 10⁻⁶. 11 CFU / g.

[0016] This embodiment also provides a method for preparing the Cistanche tubulosa health supplement that improves intestinal flora, including the following steps: S1. Cistanche tubulosa, Imperata cylindrica root, Paeonia lactiflora and Portulaca oleracea are pulverized and passed through a 100-mesh sieve to obtain Chinese herbal powder. The Chinese herbal powder is added to deionized water, soaked for 1 hour, boiled and then decocted for another hour. The mixture is filtered to obtain residue and filtrate. The ratio of Chinese herbal powder to deionized water is 1 g: 20 mL. S2. Sterilize the filtrate obtained in step S1 at 121℃ for 20 min, cool it, and then inoculate it with activated Lactobacillus paracasei, Lactobacillus reuteri and Pediococcus pentosus. Ferment at 37℃ for 48 h, centrifuge at 10000 rpm for 20 min, filter the supernatant through a 0.22 μm filter membrane, concentrate it under reduced pressure to 20% of the original volume, freeze dry it to obtain the fermentation extract; S3. Dry the filter residue obtained in step S1, add it to an 80 vol% ethanol solution, heat it under reflux in a water bath at 80°C for 1 h, centrifuge it at 8000 rpm for 20 min, remove the ethanol from the supernatant by vacuum distillation, and freeze-dry it to obtain the filter residue extract; the ratio of filter residue to ethanol solution is 1 g: 15 mL. S4. Dry the Hericium erinaceus to constant weight, pulverize it through an 80-mesh sieve to obtain Hericium erinaceus powder, add it to a 0.2 mol / L phosphate buffer solution with pH 7.8, stir at 500 rpm for 4 h, centrifuge at 10000 r / min for 20 min, and take the supernatant; the ratio of Hericium erinaceus powder to phosphate buffer solution is 1 g: 30 mL. S5. Take the supernatant obtained in step S4, add ammonium sulfate to saturation, let stand overnight at 4°C, centrifuge, place the precipitate in a 3500 Da dialysis bag and dialyze in deionized water for 24 h, freeze-dry the dialysate to obtain Hericium erinaceus water-soluble protein; S6. The Hericium erinaceus water-soluble protein obtained in step S5 was added to PBS (0.01M) buffer at pH 6.8 and stirred overnight at 4°C. The pH was adjusted to 9 with 1 mol / L NaOH. The residue extract obtained in step S3 was added and stirred for 24 h. The mixture was dialyzed in deionized water for 48 h, and the dialysate was freeze-dried to obtain the extract - Hericium erinaceus protein. The mass concentration of Hericium erinaceus water-soluble protein in PBS buffer was 25 mg / mL. S7. Take the extract obtained in step S6 - Hericium erinaceus protein, and the fermentation extract fermentation preparation obtained in step S2, fructooligosaccharides, resistant dextrin and steviol glycosides, and mix them evenly to obtain a Cistanche tubulosa health care nutritional agent that improves intestinal flora.

[0017] Example 2: A Cistanche tubulosa health supplement for improving intestinal flora, the raw materials for preparation include the following components in parts by weight: 20 parts of Cistanche tubulosa, 4 parts of Imperata cylindrica root, 2 parts of Paeonia lactiflora, 1 part of Portulaca oleracea, 2 parts of Hericium erinaceus, 4 parts of fructooligosaccharides, 2 parts of resistant dextrin, and 0.1 parts of steviol glycosides.

[0018] The Cistanche tubulosa health supplement for improving gut microbiota also includes the following components by weight: 0.05 parts of Lactobacillus paracasei, 0.03 parts of Lactobacillus reuteri, and 0.02 parts of Pediococcus pentosaceus, all in powder form, with a live bacteria count of 1.5 × 10⁻⁶. 11 CFU / g.

[0019] This embodiment also provides a method for preparing the Cistanche tubulosa health supplement that improves intestinal flora, including the following steps: S1. Cistanche tubulosa, Imperata cylindrica root, Paeonia lactiflora and Portulaca oleracea are pulverized and passed through an 80-mesh sieve to obtain Chinese herbal powder. The Chinese herbal powder is added to deionized water, soaked for 1 hour, boiled and then decocted for another hour. The mixture is filtered to obtain residue and filtrate. The ratio of Chinese herbal powder to deionized water is 1 g: 20 mL. S2. Sterilize the filtrate obtained in step S1 at 121℃ for 20 min, cool it, and then inoculate it with activated Lactobacillus paracasei, Lactobacillus reuteri and Pediococcus pentosus. Ferment at 37℃ for 48 h, centrifuge at 10000 rpm for 15 min, filter the supernatant through a 0.22 μm filter membrane, concentrate it under reduced pressure to 20% of the original volume, freeze dry it to obtain the fermentation extract; S3. Dry the filter residue obtained in step S1, add it to a 70 vol% ethanol solution, heat it under reflux in an 80°C water bath for 1 h, centrifuge it at 8000 rpm for 15 min, remove the ethanol from the supernatant by vacuum distillation, and freeze-dry it to obtain the filter residue extract; the ratio of filter residue to ethanol solution is 1 g: 10 mL. S4. Dry the Hericium erinaceus to constant weight, pulverize it through an 80-mesh sieve to obtain Hericium erinaceus powder, add it to a 0.2 mol / L phosphate buffer solution with pH 7.8, stir at 400 rpm for 4 h, centrifuge at 10000 r / min for 15 min, and take the supernatant; the ratio of Hericium erinaceus powder to phosphate buffer solution is 1 g: 20 mL. S5. Take the supernatant obtained in step S4, add ammonium sulfate to saturation, let stand overnight at 4°C, centrifuge, place the precipitate in a 3500 Da dialysis bag and dialyze in deionized water for 24 h, freeze-dry the dialysate to obtain Hericium erinaceus water-soluble protein; S6. The Hericium erinaceus water-soluble protein obtained in step S5 was added to PBS (0.01M) buffer at pH 6.8 and stirred overnight at 4°C. The pH was adjusted to 9 with 1 mol / L NaOH. The residue extract obtained in step S3 was added and stirred for 24 h. The mixture was dialyzed in deionized water for 48 h. The dialysate was freeze-dried to obtain the extract - Hericium erinaceus protein. The mass concentration of Hericium erinaceus water-soluble protein in PBS buffer was 20 mg / mL. S7. Take the extract obtained in step S6 - Hericium erinaceus protein, and the fermentation extract fermentation preparation obtained in step S2, fructooligosaccharides, resistant dextrin and steviol glycosides, and mix them evenly to obtain a Cistanche tubulosa health care nutritional agent that improves intestinal flora.

[0020] Example 3: A Cistanche tubulosa health supplement for improving intestinal flora, the raw materials for preparation include the following components in parts by weight: 25 parts of Cistanche tubulosa, 5 parts of Imperata cylindrica root, 3 parts of Paeonia lactiflora, 1.5 parts of Portulaca oleracea, 3 parts of Hericium erinaceus, 5 parts of fructooligosaccharides, 2.5 parts of resistant dextrin, and 0.15 parts of steviol glycosides.

[0021] The Cistanche tubulosa health supplement for improving gut microbiota also includes the following components by weight: 0.06 parts of Lactobacillus paracasei, 0.04 parts of Lactobacillus reuteri, and 0.025 parts of Pediococcus pentosaceus, all in powder form, with a live bacteria count of 1.5 × 10⁻⁶. 11 CFU / g.

[0022] This embodiment also provides a method for preparing the Cistanche tubulosa health supplement that improves intestinal flora, including the following steps: S1. Cistanche tubulosa, Imperata cylindrica root, Paeonia lactiflora and Portulaca oleracea are pulverized and passed through a 90-mesh sieve to obtain Chinese herbal powder. The Chinese herbal powder is added to deionized water, soaked for 1 hour, boiled and then decocted for another hour. The mixture is filtered to obtain residue and filtrate. The ratio of Chinese herbal powder to deionized water is 1 g: 20 mL. S2. Sterilize the filtrate obtained in step S1 at 121℃ for 20 min, cool it, and then inoculate it with activated Lactobacillus paracasei, Lactobacillus reuteri and Pediococcus pentosus. Ferment at 37℃ for 48 h, centrifuge at 10000 rpm for 18 min, filter the supernatant through a 0.22 μm filter membrane, concentrate it under reduced pressure to 20% of the original volume, freeze dry it to obtain the fermentation extract; S3. Dry the filter residue obtained in step S1, add it to a 75 vol% ethanol solution, heat it under reflux in a water bath at 80°C for 1 h, centrifuge it at 8000 rpm for 18 min, remove the ethanol from the supernatant by vacuum distillation, and freeze-dry it to obtain the filter residue extract; the ratio of filter residue to ethanol solution is 1 g: 12 mL. S4. Dry the Hericium erinaceus to constant weight, pulverize it through an 80-mesh sieve to obtain Hericium erinaceus powder, add it to a 0.2 mol / L phosphate buffer solution with pH 7.8, stir at 450 rpm for 4 h, centrifuge at 10000 r / min for 18 min, and take the supernatant; the ratio of Hericium erinaceus powder to phosphate buffer solution is 1 g: 25 mL. S5. Take the supernatant obtained in step S4, add ammonium sulfate to saturation, let stand overnight at 4°C, centrifuge, place the precipitate in a 3500 Da dialysis bag and dialyze in deionized water for 24 h, freeze-dry the dialysate to obtain Hericium erinaceus water-soluble protein; S6. The Hericium erinaceus water-soluble protein obtained in step S5 was added to PBS (0.01M) buffer at pH 6.8 and stirred overnight at 4°C. The pH was adjusted to 9 with 1 mol / L NaOH. The residue extract obtained in step S3 was added and stirred for 24 h. The mixture was dialyzed in deionized water for 48 h. The dialysate was freeze-dried to obtain the extract - Hericium erinaceus protein. The mass concentration of Hericium erinaceus water-soluble protein in PBS buffer was 24 mg / mL. S7. Take the extract obtained in step S6 - Hericium erinaceus protein, and the fermentation extract fermentation preparation obtained in step S2, fructooligosaccharides, resistant dextrin and steviol glycosides, and mix them evenly to obtain a Cistanche tubulosa health care nutritional agent that improves intestinal flora.

[0023] The only difference between Comparative Example 1 and Example 1 is that Imperata cylindrica root, Paeonia lactiflora, and Portulaca oleracea are not added.

[0024] The only difference between Comparative Example 2 and Example 1 is that Hericium erinaceus was not added.

[0025] The only difference between Comparative Example 3 and Example 1 is that no filter residue extract was added.

[0026] Experimental Example 1: First, prepare a 0.2 mmol DPPH ethanol solution for later use. Take 2 mL of the 0.5 mg / mL health supplement solutions from Examples 1-3 and Comparative Examples 1-3, add 2 mL of the DPPH ethanol solution, mix well, react in the dark for 30 min, and measure the absorbance at 517 nm to calculate the DPPH scavenging rate. The results are as follows: Figure 1 As shown.

[0027] Figure 1 The results showed that the DPPH scavenging rate of the health supplements in Examples 1-3 was significantly better than that in Examples 1-3. Example 1 lacked Imperata cylindrica root, Paeonia lactiflora root, and Portulaca oleracea root. These three herbs are traditional Chinese medicines rich in polyphenols, flavonoids, and organic acids, which are antioxidants. Their absence directly reduced the total amount of natural antioxidants in the system, significantly weakening its free radical scavenging ability. Example 2 lacked Hericium erinaceus (monkey head mushroom). Hericium erinaceus water-soluble protein undergoes a coupling reaction with the filter residue extract under alkaline conditions. The resulting extract-hericium erinaceus protein complex not only possesses certain antioxidant activity but may also protect the stability of small-molecule antioxidants through intermolecular interactions. The absence of this component weakened the synergistic antioxidant effect of the system. Example 3 did not add the filter residue extract, and its stability and free radical scavenging efficiency were lower than those of the complete component.

[0028] Experimental Example 2: 1% of the health supplements from Example 2 and Comparative Examples 1-3 were added to MRS medium and sterilized at 121℃ for 20 min. The activated Bifidobacterium longum and Lactobacillus acidophilus were inoculated into the medium at a 5% inoculum size. The medium was incubated at 37℃ with shaking for 48 h. Samples were taken at 0 h and 48 h, and diluted 10 μL with physiological saline. 6 100 μL of the culture medium was spread evenly on a solid plate using a spread plate method and incubated at 37°C for 48 h. The number of viable bacteria per milliliter of culture was determined by plate count. The difference in viable bacterial count between 48 h and 0 h was calculated to obtain the change in probiotics. The results are as follows: Figure 2 As shown.

[0029] Figure 2The results show the growth-promoting effects of the health supplements in Examples 2 and 1-3 on *Bifidobacterium longum* and *Lactobacillus acidophilus*. Example 2 showed a significantly higher increase in viable *Bifidobacterium longum* and *Lactobacillus acidophilus* compared to Comparative Examples 1-3. This indicates that the nutrient supplement of the present invention can provide probiotics with abundant fermentable carbon sources and growth-promoting factors; the fermentation extract contains microbial metabolites and active bacterial components, exhibiting growth-promoting activity; and the extract-Hericium erinaceus protein conjugate may maintain the continuous proliferation of probiotics through the sustained release of small molecule active substances. The absence of several traditional Chinese medicines in Comparative Example 1 reduced the source of water-soluble polysaccharides and oligosaccharides, decreasing the types and total amount of carbon sources available to probiotics. The growth-promoting effect of Comparative Example 2 was further reduced. After the absence of the Hericium erinaceus protein conjugate, some of the active small molecules bound to the protein were rapidly released and diluted in the stomach and upper intestine, resulting in insufficient effective concentrations reaching the probiotic enrichment zone in the colon. Comparative Example 3, lacking the small molecule extract from the filter residue, could not effectively support the continuous proliferation of probiotics.

[0030] Experiment 3: After one week of acclimatization, mice were randomly divided into four groups of eight mice each: a blank control group, a model group, Example 1 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group. The model group, Example 1 group, and Comparative Examples 1-3 groups were administered cefixime via gavage to induce intestinal flora imbalance at a dose of 30 mg / kg, administered daily at noon. Example 1 and Comparative Examples 1-3 groups were administered a health supplement twice daily via gavage at a dose of 200 mg / kg, with a 3-hour interval between administrations. The control group and model group received an equal volume of physiological saline. Body weight was measured on days 1, 3, 6, 9, 12, and 14. The results are as follows: Figure 3 As shown.

[0031] Figure 3 This study investigated the weight changes of a mouse model with antibiotic-induced gut microbiota dysbiosis over 14 days after administration of different nutrient solutions. In the model group, mice showed a significant decrease in weight after gavage with cefixime, indicating that cefixime successfully induced gut microbiota dysbiosis, leading to impaired digestion and absorption, stagnant or even decreased weight gain. In Example 1, mice showed some weight loss after gavage with the nutrient solution of this invention, with their weight approaching that of the control group on day 14. This was attributed to the synergistic effect of the fermented extract, extract-heric mane mushroom protein conjugate, compound probiotics, and prebiotics in the nutrient solution—probiotics and prebiotics promote the proliferation of beneficial gut bacteria, the protein conjugate protects and slowly releases the active ingredients, and the fermented extract provides post-biotic substances, collectively repairing the damaged intestinal barrier and restoring the body's nutrient absorption function. In Comparative Example 1, the weight recovery was significantly weaker than in Example 1, with the final weight falling between the model group and Example 1, indicating that the absence of the traditional Chinese medicine components weakened the overall formula's repair effect on intestinal damage. The weight recovery trend in Comparative Example 2 was declining, suggesting that the protective and sustained-release effects of Hericium erinaceus protein conjugates on the active ingredients significantly contribute to maintaining the sustainability of intestinal repair. The final weight of Comparative Example 3 was lower than that of Example 1.

[0032] Those skilled in the art should understand that the discussion of any of the above embodiments is merely exemplary and is not intended to imply that the scope of the invention is limited to these examples; within the framework of the invention, the technical features of the above embodiments or different embodiments can also be combined, the steps can be implemented in any order, and there are many other variations of the different aspects of the invention as described above, which are not provided in detail for the sake of brevity.

Claims

1. A health supplement containing Cistanche tubulosa that improves intestinal flora, characterized in that, The raw materials for preparation include the following components in parts by weight: 20-30 parts of Cistanche tubulosa, 4-6 parts of Imperata cylindrica root, 2-4 parts of Paeonia lactiflora, 1-2 parts of Portulaca oleracea, 2-4 parts of Hericium erinaceus, 4-6 parts of fructooligosaccharides, 2-3 parts of resistant dextrin, and 0.1-0.2 parts of steviol glycosides.

2. The Cistanche tubulosa health supplement for improving intestinal flora according to claim 1, characterized in that, The raw materials also include the following components in parts by weight: 0.05-0.08 parts of *Lactobacillus paracasei*, 0.03-0.05 parts of *Lactobacillus reuteri*, and 0.02-0.03 parts of *Pediococcus pentosaceus*, all of which are powder preparations with a viable count of 1.5 × 10⁻⁶. 11 CFU / g.

3. A method for preparing a Cistanche tubulosa health supplement for improving intestinal flora as described in any one of claims 1-2, characterized in that, Includes the following steps: S1. Cistanche tubulosa, Imperata cylindrica root, Paeonia lactiflora and Portulaca oleracea are pulverized through an 80-100 mesh sieve to obtain Chinese herbal powder. The Chinese herbal powder is added to deionized water, soaked for 1 hour, boiled and then decocted for another 1 hour. The mixture is filtered to obtain residue and filtrate. S2. Sterilize the filtrate obtained in step S1 at 121℃ for 20 min, cool it, and then inoculate it with activated Lactobacillus paracasei, Lactobacillus reuteri and Pediococcus pentosus. Ferment at 37℃ for 48 h, centrifuge at 10000 rpm for 15-20 min, filter the supernatant through a 0.22 μm filter membrane, concentrate it under reduced pressure to 20% of the original volume, and freeze dry to obtain the fermentation extract. S3. Dry the filter residue obtained in step S1, add it to a 70-80 vol% ethanol solution, heat it under reflux in an 80°C water bath for 1 h, centrifuge it at 8000 rpm for 15-20 min, remove the ethanol from the supernatant by vacuum distillation, and freeze-dry it to obtain the filter residue extract. S4. Dry the Hericium erinaceus to constant weight, pulverize it through an 80-mesh sieve to obtain Hericium erinaceus powder, add it to a 0.2 mol / L phosphate buffer solution at pH 7.8, stir at 400-500 rpm for 4 h, centrifuge at 10000 r / min for 15-20 min, and take the supernatant. S5. Take the supernatant obtained in step S4, add ammonium sulfate to saturation, let stand overnight at 4°C, centrifuge, place the precipitate in a 3500 Da dialysis bag and dialyze in deionized water for 24 h, freeze-dry the dialysate to obtain Hericium erinaceus water-soluble protein; S6. The Hericium erinaceus water-soluble protein obtained in step S5 was added to PBS (0.01M) buffer at pH 6.8 and stirred overnight at 4°C. The pH was adjusted to 9 with 1 mol / L NaOH. The filter residue extract obtained in step S3 was added and stirred for 24 h. The mixture was dialyzed in deionized water for 48 h. The dialysate was freeze-dried to obtain the extract - Hericium erinaceus protein. S7. Take the extract obtained in step S6 - Hericium erinaceus protein, and the fermentation extract fermentation preparation obtained in step S2, fructooligosaccharides, resistant dextrin and steviol glycosides, and mix them evenly to obtain a Cistanche tubulosa health care nutritional agent that improves intestinal flora.

4. The preparation method of the Cistanche tubulosa health supplement for improving intestinal flora according to claim 3, characterized in that, In step S1, the ratio of the traditional Chinese medicine powder to deionized water is 1 g: 20 mL.

5. The preparation method of the Cistanche tubulosa health supplement for improving intestinal flora according to claim 4, characterized in that, In step S3, the ratio of the filter residue to the ethanol solution is 1 g: 10-15 mL.

6. The preparation method of the Cistanche tubulosa health supplement for improving intestinal flora according to claim 5, characterized in that, In step S4, the ratio of the Hericium erinaceus powder to the phosphate buffer is 1 g: 20-30 mL.

7. The preparation method of the Cistanche tubulosa health supplement for improving intestinal flora according to claim 6, characterized in that, In step S6, the mass concentration of the Hericium erinaceus water-soluble protein in the PBS buffer is 20-25 mg / mL.

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