Lactobacillus gasseri CCFM1481 for converting curcumin to produce tetrahydrocurcumin and enhance anti-fatigue effect

Abstract

The invention discloses lactobacillus gasseri CCFM1481 for converting curcumin to produce tetrahydrocurcumin and enhance an anti-fatigue effect, and belongs to the technical field of microorganisms. The lactobacillus gasseri CCFM1481 obtained through screening has the capacity of converting curcumin, curcumin can be utilized, tetrahydrocurcumin can be generated, and the conversion rate is 89.83%. The invention further provides a synbiotic preparation prepared from the lactobacillus gasseri CCFM1481 and curcumin, and the synbiotic preparation can be used as an anti-fatigue functional medicine and has a good application prospect and an industrialization foundation.

Description

Technical Field

[0001] This invention relates to a strain of Lactobacillus gasseri CCFM1481 that transforms curcumin to produce tetrahydrocurcumin, thereby enhancing its anti-fatigue effects, and belongs to the field of microbial technology. Background Technology

[0002] Fatigue is a typical manifestation of sub-health, leading to a series of physiological changes in the body. Fatigue usually occurs during strenuous and sustained physical activity. Currently, fatigue is mainly classified into physiological and psychological fatigue. Physiological fatigue is further divided into central and peripheral fatigue based on its location. Central fatigue can cause a series of fatigue-like reactions in the body, nervous system, and mind, often occurring after strenuous exercise, and is characterized by significant changes in the synthesis and metabolism of neurotransmitters during exercise. Peripheral fatigue is caused by decreased muscle function after excessive activity, mainly involving factors such as energy supply and the accumulation of metabolites. Psychological fatigue is caused by mental activity; prolonged monotonous and repetitive work and life can reduce enthusiasm and interest in work and life. Fatigue can exacerbate primary diseases, having a significant impact on the patient's physiology and psychology, and seriously affecting their quality of life.

[0003] Traditional Chinese medicine provides reliable clinical practice for preventing and treating fatigue and alleviating secondary diseases. It boasts a rich variety of natural drugs with anti-fatigue effects, offering a new field for the development and utilization of novel anti-fatigue drugs. Curcumin, a safe and non-toxic natural diketone pigment, is widely used in food production. The significant and diverse health effects of curcumin have been widely reported, which contrasts sharply with its poor physical stability, bioavailability, and pharmacokinetic characteristics. Tetrahydrocurcumin, as the main active metabolite of curcumin, exhibits slower metabolism in vivo due to its altered chemical structure, allowing it to maintain its activity for a longer period and possessing higher bioavailability and stability than curcumin. It can reduce oxidative stress and inhibit free radical production by activating antioxidant enzyme systems, proving more effective than curcumin, and its broad pharmacological effects involve regulating various signal transduction pathways. Therefore, obtaining strains capable of metabolizing curcumin to produce tetrahydrocurcumin and enhancing its anti-fatigue efficacy, and applying it to food, health products, or pharmaceuticals, is of great significance. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a strain of *Lactobacillus gasseri* capable of converting curcumin to enhance its anti-fatigue effects. Lactobacillus gasseri CCFM1481 was mentioned, and its application was provided.

[0005] This invention provides a strain of Lactobacillus gasseri ( Lactobacillus gasseri CCFM1481 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC No: 66129.

[0006] This invention provides a product containing the aforementioned Lactobacillus gasseri ( Lactobacillus gasseri CCFM1481 microbial preparation.

[0007] In one embodiment, the content of Lactobacillus gasseri CCFM1481 in the microbial preparation is not less than 1×10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0008] In one embodiment, the microbial preparation is a solid or liquid preparation.

[0009] The present invention also provides a synbiotic preparation containing the aforementioned Lactobacillus gasseri CCFM1481.

[0010] In one embodiment, the synbiotic preparation contains Lactobacillus gasseri CCFM1481 and its fermentation products.

[0011] In one embodiment, the curcumin ferment contains tetrahydrocurcumin.

[0012] In one embodiment, the fermentation product is obtained by fermentation in a curcumin-containing system.

[0013] In one embodiment, the preparation of the fermentation product includes the following steps: (1) Ferment Lactobacillus gasseri CCFM1481 in a fermentation substrate containing curcumin; (2) Centrifuge the fermentation broth obtained in step (1), collect the supernatant, and obtain curcumin fermentation product.

[0014] In one embodiment, step (1) involves feeding Lactobacillus gasseri CCFM1481 at a concentration of not less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 The fermentation system was inoculated with a concentration of CFU / g.

[0015] In one implementation, the method includes the following steps: (1) Inoculate Lactobacillus gasseri CCFM1481 into the culture medium and culture at 35-40℃ for 48 h to obtain bacterial culture; (2) Add the bacterial solution obtained in step (1) to the fermentation substrate containing curcumin at an inoculation rate of 5% (v / v), ferment at 25-40℃ for 48h, and collect the fermentation supernatant.

[0016] The present invention also provides curcumin ferments prepared using the method described herein.

[0017] The present invention provides a composition containing the Lactobacillus gasseri CCFM1481 and curcumin.

[0018] The present invention also provides a probiotic product containing the Lactobacillus gasseri CCFM1481 or the curcumin ferment.

[0019] In one embodiment, the product is food, medicine, or health product.

[0020] In one embodiment, the food is a dairy product, soy product, or fruit and vegetable product prepared using at least one of the curcumin ferment, synbiotic preparation, Lactobacillus gasseri CCFM1481, or microbial preparation.

[0021] In one embodiment, the dairy products include fermented milk, flavored fermented milk, fermented milk beverages, cream, cheese, milk-containing beverages, or milk powder; the soy products include soy milk and soy milk powder; and the fruit and vegetable products include fruit and vegetable products made from at least one of cabbage, white radish, cucumber, beet, yellow peach, or bayberry products.

[0022] In one embodiment, the food is a fermented food, including solid food, liquid food, or semi-solid food.

[0023] In one embodiment, the food is a beverage or snack containing at least one of the above-mentioned curcumin ferment, Lactobacillus gasseri CCFM1481, or microbial preparations.

[0024] In one embodiment, the drug further comprises a drug carrier and / or pharmaceutical excipients.

[0025] In one embodiment, the carrier includes one or more of the following commonly used in medicine: fillers, adhesives, wetting agents, disintegrants, lubricants, and flavoring agents.

[0026] In one embodiment, the dosage form of the drug is granules, capsules, tablets, pills, or oral liquid.

[0027] The present invention also provides the use of the Lactobacillus gasseri CCFM1481, the curcumin ferment, or the synbiotic preparation in the preparation of a drug with anti-fatigue function.

[0028] In one embodiment, the product has at least one of the following functions (1)-(4): (1) Increase individual exercise time; (2) Reduces the accumulation of serum metabolites (LA, BUN) in individuals; (3) Increase the expression of fatigue-related genes (DRD1, DBH, MCT1, LDHB) in the body.

[0029] Beneficial effects: 1. The Lactobacillus gasseri CCFM1481 provided by this invention has the ability to convert curcumin. After fermentation in a curcumin-containing fermentation substrate, the degradation rate of curcumin is 95%, and tetrahydrocurcumin active substance is generated, with a conversion rate of 89.83%.

[0030] 2. The Lactobacillus gasseri CCFM1481 provided by this invention can promote the efficacy of curcumin, specifically in the following ways: (1) The synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin can increase the swimming time of mice, extending the exercise time from 9.18±0.97 minutes to 15.05±1.65 minutes and 16.20±2.25 minutes, respectively.

[0031] (2) The synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin can improve the open field behavior of mice. The open field movement distance increased by 23.98% and 33.24% respectively compared with the curcumin group, and the open field movement time increased by 31.64% and 31.69% respectively compared with the curcumin group.

[0032] (3) The synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin can improve the ability to clear serum metabolites. The LA level was reduced by 17.94% and 14.09% respectively compared with the curcumin group, and the BUN level was reduced by 25.46% and 28.46% respectively.

[0033] (4) Fermentation of curcumin by Lactobacillus gasseri CCFM1481 can increase the expression of anti-fatigue related genes. The expression of DRD1 gene increased by 35.98% compared with the curcumin group, and the expression level of DBH gene increased by 30.75% compared with the curcumin group.

[0034] Anti-fatigue products can be produced using Lactobacillus gasseri CCFM1481 and curcumin. The production process utilizes curcumin as a raw material, enhancing its efficacy. This process boasts advantages such as safety, high efficiency, low cost, and mild reaction, making it suitable for large-scale industrial production. Therefore, the application of Lactobacillus gasseri CCFM1481 to curcumin-containing products has enormous application potential and a solid foundation.

[0035] Preservation of biological materials Lactobacillus gasseri CCFM1481, taxonomically named Lactobacillus gasseri It was deposited on April 11, 2025 at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC No: 66129, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0036] Figure 1 High-performance liquid chromatography (HPLC) chromatograms of curcumin before and after fermentation; Figure 2 LC-MS images of curcumin before and after fermentation; Figure 3 LC-MS images of tetrahydrocurcumin before and after fermentation; Figure 4 Time to exhaustion during swimming for mice in different groups; Figure 5 : Open field experiment diagrams of behavioral characteristics in mice of different groups; Figure 6 Serum metabolite levels in mice from different groups; Figure 7 Expression levels of neuroprotective genes in mice from different groups; Figure 8 Expression levels of genes related to metabolite clearance in mice of different groups. Detailed Implementation

[0037] The SPF-grade male C57BL / 6J mice (6 weeks old, 20±2 g) used in the following examples were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.; the curcumin used in the following examples was purchased from Maclean's Reagent Company; chloroform, isopropanol, ethanol, TRIzol, DEPC-treated water, and sodium chloride were from China National Pharmaceutical Reagent Co., Ltd.; lead wire was from Mingpinhui Hardware Store; grinding beads were from Huzhou Xiongsheng Grinding Co., Ltd.; blood urea nitrogen (BUN) and blood lactate (BLA) were from Nanjing Jiancheng Bioengineering Institute; the BCA protein concentration assay kit was from Shanghai Beyotime Biotechnology Co., Ltd.; the reverse transcription kit and real-time fluorescence quantitative kit were from Nanjing Novizan Biotechnology Co., Ltd.; and the primers were from Shanghai Sangon Biotech Co., Ltd.

[0038] The culture media involved in the following examples are as follows: MRS solid medium: peptone 10 g / L, beef extract 10 g / L, glucose 20 g / L, sodium acetate 2 g / L, yeast extract 5 g / L, diammonium hydrogen citrate 2 g / L, K2PO4·3H2O 2.6 g / L, MgSO4·7H2O 0.1 g / L, MnSO4 0.05 g / L, Tween 1 mL / L, agar 20 g / L.

[0039] MRS liquid culture medium: peptone 10 g / L, beef extract 10 g / L, glucose 20 g / L, sodium acetate 2 g / L, yeast extract 5 g / L, diammonium hydrogen citrate 2 g / L, K2PO4·3H2O 2.6 g / L, MgSO4·7H2O 0.1 g / L, MnSO4 0.05 g / L, Tween 1 mL / L.

[0040] Curcumin MRS liquid culture medium: peptone 10 g / L, beef extract 10 g / L, glucose 20 g / L, sodium acetate 2 g / L, yeast extract 5 g / L, diammonium hydrogen citrate 2 g / L, K2PO4·3H2O 2.6 g / L, MgSO4·7H2O 0.1 g / L, MnSO4 0.05 g / L, Tween 1 mL / L, curcumin 100 mg / L.

[0041] Fermentation substrate containing curcumin: curcumin 100 mg / L, peptone 10 g / L, beef extract 10 g / L, glucose 20 g / L, sodium acetate 2 g / L, yeast powder 5 g / L, diammonium hydrogen citrate 2 g / L, K2PO4·3H2O 2.6 g / L, MgSO4·7H2O 0.1 g / L, MnSO4 0.05 g / L, Tween mL / L.

[0042] The detection methods involved in the following embodiments are as follows: Method for detecting curcumin: Qualitative and quantitative analysis was performed using an Agilent 1260 Infinity III liquid chromatograph. A C18 column (250 × 4.6 mm, 5 µm) was used, with an autosampler performing gradient elution at a flow rate of 1.00 mL / min. The column temperature was 30℃, and the injection volume was 10 μL. The UV detector wavelength was 420 nm. The mobile phase was: A - 0.2% formic acid in water, B - acetonitrile; gradient elution: 0–6 min 20% B, 6–15 min 20%–35% B, 15–25 min 35%–70% B, 25–35 min 70%–40% B, 35–40 min 40%–20% B.

[0043] Example 1: Screening, strain identification and preservation of Lactobacillus gasseri CCFM1481 1. Screening Using healthy adult feces as samples, the samples were serially diluted 10-fold with sterile saline to 10⁻⁶. -4 10 -5 10 -6 Then take 100 μL of each diluted by a factor of 10. -4 10 -5 10 -6The diluted solution was plated on MRS solid medium and incubated at 37°C for 48 h. The colony morphology was observed and recorded. Colonies of different morphologies were picked from the MRS solid medium and streaked for isolation. After incubation at 37°C for 48 h, single colonies of different morphologies were picked from the MRS solid medium again and streaked for isolation until pure single colonies with consistent morphology were obtained. Pure colonies from the MRS solid medium were inoculated into 5 mL of MRS liquid medium and incubated at 37°C for 24 h. 1 mL of bacterial solution was taken into a sterile centrifuge tube, centrifuged at 8000 r / min for 3 min, and the upper medium was discarded. The obtained bacterial sludge was freeze-dried.

[0044] 2. Identification Morphological, physiological and biochemical, and molecular biological identifications were performed on the selected strains, confirming that the strain was classified as Lactobacillus gasseri and named Lactobacillus gasseri CCFM1481.

[0045] 3. Save Lactobacillus gasseri CCFM1481 was inoculated into 5 mL of MRS liquid medium and cultured at 37°C for 24 h. 1 mL of bacterial culture was placed in a sterile centrifuge tube, centrifuged at 8000 r / min for 3 min, and the upper culture medium was discarded. The bacterial sludge was resuspended in 30% glycerol solution and stored at -80°C.

[0046] Example 2: Preparation of a synergistic formulation of Lactobacillus gasseri CCFM1481 and curcumin, and curcumin fermentation product 1. Preparation of Lactobacillus gasseri CCFM1481-curcumin ferment: (1) The Lactobacillus gasseri CCFM1481 in Example 1 was streaked on MRS solid medium, and the plate was incubated upside down at 37°C for 48 h. A single colony was picked and inoculated into 5 mL of MRS liquid medium and incubated at 37°C for 48 h to obtain Lactobacillus gasseri CCFM1481 bacterial suspension. (2) Add 5% (v / v) of Lactobacillus gasseri CCFM1481 culture obtained in step (1) to the curcumin-containing fermentation substrate, and ferment at 37℃ for 48 h. The initial concentration of the culture is 1×10⁻⁶. 6 CFU / mL; (3) After fermentation, centrifuge at 8000 r / min for 20 min, collect the supernatant, filter it through a 0.22 μm disposable filter for sterilization, dispense it into portions, and store it in a -20℃ refrigerator to obtain the curcumin ferment of Lactobacillus gasseri CCFM1481. The content of tetrahydrocurcumin was determined to be 73.87 μg / mL.

[0047] 2. Preparation of Lactobacillus gasseri CCFM1481-curcumin synergistic preparation: Lactobacillus gasseri CCFM1481 from Example 1 was streaked onto MRS solid medium and incubated upside down at 37°C for 48 h. A single colony was picked and inoculated into 5 mL of MRS liquid medium and incubated at 37°C for 48 h to obtain a Lactobacillus gasseri CCFM1481 bacterial suspension. The suspension was centrifuged to collect the bacterial sludge, which was then resuspended in a 10 g / L curcumin solution to obtain a bacterial concentration of 5 × 10⁻⁶. 9 CFU / mL Lactobacillus gasseri CCFM1481-curcumin synergistic preparation.

[0048] Example 3: Conversion of curcumin by Lactobacillus gasseri CCFM1481 1. Streaked Lactobacillus gasseri CCFM1481 from Example 1 onto MRS solid medium and incubated upside down at 37°C for 48 h; pick a single colony and inoculate it into 5 mL of MRS liquid medium and incubate at 37°C for 48 h.

[0049] 2. Add 5% (v / v) of Lactobacillus gasseri CCFM1481 culture obtained in step 1 to the curcumin-containing fermentation substrate, and ferment at 37℃ for 48 h. The initial concentration of the culture is 1×10⁻⁶. 6 CFU / mL.

[0050] 3. After fermentation, the fermentation supernatant was filtered through a 0.22 μm single-use microporous membrane and stored at 4℃ for later use.

[0051] 4. Accurately weigh 10 mg of curcumin reference standard into a 10 mL volumetric flask, accurately pipette the above solution and dilute to 150 μg / mL, 100 μg / mL, 50 μg / mL, 25 μg / mL and 5 μg / mL, and plot a working curve under mobile phase.

[0052] 5. Take 2 mL of the fermentation product obtained in step (3) and the control group solution (fermentation substrate containing curcumin), and analyze the pre-fermentation state using HPLC under chromatographic conditions. Figure 1 A) and after fermentation ( Figure 1 B), the reduction rate of curcumin was 95%.

[0053] 6. Take 2 mL of the fermentation product obtained in step (3) and the control group solution, and analyze the fermentation process before and after fermentation by LC-MS. Figure 2 The content of curcumin (AB) was measured, and the results showed that while curcumin decreased, tetrahydrocurcumin was generated. The content of tetrahydrocurcumin before and after fermentation was measured. Figure 3 (AB), and the amount of metabolites generated was calculated to be 73.87 μg / mL, with a conversion rate of 89.83%.

[0054] Example 4: Effects of a synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin on swimming time in fatigued mice The fermented product of Lactobacillus gasseri CCFM1481-curcumin and the synergistic preparation of Lactobacillus gasseri CCFM1481-curcumin were prepared according to the steps of Example 2.

[0055] Forty-eight healthy male C57BL / 6J mice aged 6 weeks were randomly divided into 6 groups, with 8 mice in each group. The 6 groups were: blank control group (NC), model group (MC), curcumin group (CUR), Lactobacillus gasseri CCFM1481 (CCFM1481), Lactobacillus gasseri CCFM1481-curcumin synthesis group (CCFM1481 compound CUR), and Lactobacillus gasseri CCFM1481 curcumin fermentation group (CCFM1481 fermented CUR). After acclimatization for 1 week, weeks 2-4 were treated with gavage intervention, with consistent daily gavage times, and forced swimming (6 min / day) and forced restraint (4 h / day). Behavioral training was conducted in week 5. The NC and MC groups were administered saline by gavage, the CUR group was administered 100 mg / kg of mouse body weight by gavage, and the CCFM1481 group was administered a bacterial concentration of 5 × 10⁻⁶ mg / kg of mouse body weight by gavage. 9 CCFM1481 bacterial suspension at CFU / mL, CCFM1481 combined with CUR group at 5×10 9 CFU / mL and 100 mg / kg curcumin were mixed and administered by gavage; the CCFM1481 fermented CUR group was administered Lactobacillus CCFM1481 curcumin fermentation product by gavage. The amount of curcumin in the fermentation substrate containing curcumin before fermentation was converted to the amount of curcumin fermentation product corresponding to 100 mg curcumin / kg body weight.

[0056] Specific methods for behavioral training: An open field behavior detector with four grids was used for open field testing. Each mouse was placed in the middle grid, and its movement trajectory within the grid was recorded for 6 minutes. The movement of each mouse was also recorded using an animal behavior video tracking system. Training continued for 4 days.

[0057] The experimental animals were grouped as shown in Table 1.

[0058] Table 1 Grouping of experimental animals

[0059] On day 5 of week 5, the swimming exhaustion time of mice was measured: mice were placed in a swimming tank with a water depth of 25 cm and a water temperature of 25-30°C, with a lead weight of 6% of their body weight attached to their tails. Swimming exhaustion was defined as the time a mouse's head remained submerged for more than 5 seconds. The time from the start of swimming to exhaustion was recorded as the swimming exhaustion time. 24 hours later, mice were allowed to swim without weight for 5 minutes, after which they were sacrificed, and serum and muscle tissue were collected for relevant indicator measurements.

[0060] Experimental results are as follows Figure 4 As shown, compared with the NC group (21.85±2.84 minutes), the swimming time of mice in the MC group (9.18±0.97 minutes) was significantly shortened. The CUR group extended the exercise time to 11.19±1.16 minutes, while the CCFM1481 compound CUR group and the CCFM1481 fermented CUR group extended the exercise time to 15.05±1.65 minutes and 16.20±2.25 minutes, respectively. These results indicate that the synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin, as well as fermented curcumin, improved the ability of mice to exercise for longer periods.

[0061] Example 5: Effects of a synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin on open field behavior in fatigued mice The grouping, modeling, and treatment methods for C57BL / 6J mice are the same as in Example 4.

[0062] On week 5, day 5, the open field test was performed on mice: an open field behavior detector with four grids was used for the open field test. The movement of each mouse was recorded using an animal behavior video tracking system. The mice's movement trajectory within the activity box was recorded using OFT (Out-of-Flight Theory).

[0063] Experimental results are as follows Figure 5As shown, compared with the NC group, the open field movement distance of mice in the MC group (1245.08±323.86cm) was significantly reduced, while the CUR group (1736.34±151.19cm) increased by 39.468% compared with the MC group. The CCFM1481 compound CUR group (2152.68±917.15cm) and the CCFM1481 fermented CUR group (2313.45±350.39cm) increased by 72.89% and 85.81% respectively compared with the MC group. Compared with the NC group, the open field movement time ratio of mice in the MC group (41.21±6.18%) was significantly decreased. Compared with the MC group, the CUR group (54.19±5.18%) was increased by 31.49%, while the CCFM1481 compound CUR group (71.34±13.20%) and the CCFM1481 fermented CUR group (71.36±3.50%) were increased by 73.10% and 73.17% respectively compared with the MC group. These results indicate that the synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin are more effective than curcumin in relieving mental fatigue in mice.

[0064] Example 6: Effects of a synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin on serum metabolite accumulation in fatigued mice The grouping, modeling, and treatment methods for C57BL / 6J mice are the same as in Example 4.

[0065] Mice were anesthetized and blood was collected from the orbital cavity. 3000 ml of blood samples were collected. g Centrifuge for 15 min and collect the supernatant. Determine the levels of lactate (LA) and blood urea nitrogen (BUN) in mouse serum according to the kit instructions.

[0066] BUN (Brain Urine) is a byproduct of energy metabolism and a sensitive indicator related to fatigue. LA (Lactate) is a major byproduct of anaerobic metabolism, and the accumulation of lactate in serum is a significant cause of fatigue. Experimental results are as follows... Figure 6As shown, compared with the NC group, the serum LA level of mice in the MC group was significantly increased. Compared with the MC group (12.51±1.48 mmol / L), the LA level in the CUR group (10.55±1.01 mmol / L) decreased by 15.69%, while the LA levels in the CCFM1481 compound CUR group (8.66±1.74 mmol / L) and the CCFM1481 fermented CUR group (9.06±1.29 mmol / L) decreased by 30.80% and 27.55%, respectively. Compared with the NC group, the BUN level in the MC group was significantly increased. Compared with the MC group (11.68±1.47 mmol / L), the BUN level in the CUR group (9.99±0.58 mmol / L) decreased by 14.45%, while the BUN levels in the CCFM1481 compound CUR group (7.45±1.30 mmol / L) and the CCFM1481 fermented CUR group (7.15±2.06 mmol / L) decreased by 36.25% and 38.81%, respectively. These results indicate that fatigue-induced exercise can significantly increase the levels of serum metabolites LA and BUN in the blood, and the synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin, as well as fermented curcumin, are more effective than curcumin in alleviating the increase in blood LA and BUN levels caused by exercise fatigue.

[0067] Example 7: Effects of a synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin on the expression of neuroprotective genes in fatigued mice. The grouping, modeling, and treatment methods for C57BL / 6J mice are the same as in Example 4.

[0068] Mice were euthanized by decapitation. The scalp was immediately cut open on ice, and the skull and dura mater were removed. Brain tissue was carefully separated, weighed, and then mixed with physiological saline at a 1:9 (w / v) ratio. The tissue was then homogenized in a high-throughput tissue homogenizer under ice bath conditions. The resulting homogenate was centrifuged at 12000 g for 15 min at 4°C, and the supernatant was collected. To correct for the degree of homogenization, the protein concentration of the supernatant was determined using a BCA protein concentration assay kit. Total RNA was extracted using TRIzol, and cDNA was obtained by reverse transcription using a reverse transcription kit. The mRNA expression level of the gene was detected by RT-qPCR using a real-time quantitative PCR kit. The primer sequences used are shown in the table.

[0069] Table 2 Primer Sequences

[0070] Experimental results are as follows Figure 7As shown, compared with the NC group, the gene expression level of DRD1 in the MC group was significantly decreased to 0.52±0.19. Compared with the MC group, the gene expression level of CUR group (0.76±0.06) increased by 45.38%, while the gene expression levels of CCFM1481 compound CUR group (0.80±0.10) and CCFM1481 fermented CUR group (1.03±0.29) increased by 53.85% and 97.69%, respectively. Compared with the NC group, the gene expression of DBH in the MC group was significantly decreased to 0.53±0.10. Compared with the MC group, the gene expression level of CUR group (0.72±0.10) increased by 19.20%, while the gene expression levels of CCFM1481 compound CUR group (0.75±0.11) and CCFM1481 fermented CUR group (0.94±0.35) increased by 22.20% and 41.40%, respectively. These results indicate that the synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin, as well as fermented curcumin, can relieve fatigue by upregulating the expression levels of neuroprotective genes.

[0071] Example 8: Effects of a synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin and fermented curcumin on the expression of serum metabolite clearance genes in fatigued mice. The grouping, modeling, and treatment methods for C57BL / 6J mice were the same as in Example 4. The RT-qPCR method was the same as in Example 7.

[0072] Collect skeletal muscle (gastrocnemius) tissue from both lower legs of mice, wash with physiological saline, blot dry with filter paper, weigh, add physiological saline at a ratio of 1:9 (w / v), place in a high-throughput tissue homogenizer under ice bath conditions for tissue disruption, centrifuge the resulting homogenate at 12000 g for 15 min at 4℃, and collect the supernatant.

[0073] Experimental results are as follows Figure 8 As shown, compared with the NC group, the LDHB gene expression level in the MC group was significantly reduced to 0.45±0.11. Compared with the MC group, the CUR gene expression level was upregulated to 0.71±0.13, and the CCFM1481 compound CUR group and the CCFM1481 fermented CUR group were upregulated to 0.75±0.13 and 0.91±0.18, respectively. Compared with the NC group, the MCT1 gene expression level in the MC group was significantly reduced to 0.48±0.14. Compared with the MC group, the CUR gene expression level was increased to 0.82±0.22, and the CCFM1481 compound CUR group and the CCFM1481 fermented CUR group were increased to 0.86±0.20 and 0.94±0.27, respectively. These results indicate that the synergistic preparation of Lactobacillus gasseri CCFM1481 and curcumin, as well as fermented curcumin, can alleviate fatigue by regulating the expression levels of LDHB and MCT1 genes.

[0074] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. Lactobacillus gasseri ( Lactobacillus gasseri CCFM1481 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 11, 2025, with accession number GDMCC No: 66129.

2. A microbial preparation containing Lactobacillus gasseri CCFM1481 as described in claim 1.

3. The microbial preparation according to claim 2, characterized in that, The content of Lactobacillus gasseri CCFM1481 in the microbial preparation is not less than 1×10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

4. A synbiotic preparation containing Lactobacillus gasseri CCFM1481 as described in claim 1.

5. The synthetic preparation according to claim 4, characterized in that, It contains the Lactobacillus gasseri CCFM1481 and its fermentation products; the fermentation products are obtained by fermentation in a system containing curcumin.

6. The synthetic preparation according to claim 5, characterized in that, The preparation of the fermentation product includes the following steps: (1) Ferment the Lactobacillus gasseri CCFM1481 in a fermentation substrate containing curcumin; (2) Centrifuge the fermentation broth obtained in step (1), collect the supernatant, and obtain curcumin fermentation product.

7. The composition, characterized in that, It contains Lactobacillus gasseri CCFM1481 as described in claim 1 and curcumin.

8. The composition according to claim 7, characterized in that, The composition is a food, medicine, or health product.

9. The use of Lactobacillus gasseri CCFM1481 as described in claim 1, the curcumin ferment, or the synbiotic preparation in the preparation of a drug with anti-fatigue function.

10. The application according to claim 9, characterized in that, The drug has at least one of the following effects: (1) Increase individual exercise time; (2) Reduces the accumulation of serum metabolites in individuals; (3) Increase the expression of fatigue-related genes in the body.

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