Humanized probiotic strain and application thereof in preparation of medicine for preventing or treating acne

Abstract

The invention discloses a humanized probiotic strain and an application of the humanized probiotic strain in preparation of a medicine for preventing or treating acne, the humanized probiotic strain is Lactobacillus rhamnosus GG SMUDH Yang02, the preservation unit is called as GDMCC for short, the preservation number is GDMCC No: 65950, the humanized probiotic strain is Lacticasei paracasei (LP) YANG03, the preservation unit is called as GDMCC for short, and the preservation number is GDMCC No: 65951. The protective strain deleted by the acne patient is screened and separated from the intestinal tract of a healthy person, the strain can normally survive in the intestinal tract of the human body, no toxic or side effect exists, and a new way is provided for prevention and treatment of the acne patient by supplementing the deleted protective strain and active metabolite of the patient in a targeted mode.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of probiotics, in particular to probiotics for preventing and treating acne. BACKGROUND

[0002] Acne vulgaris is a chronic inflammatory, recurrent, and disfiguring skin disease involving the pilosebaceous unit, which is prone to cause facial pigmentation and even scar formation in patients, seriously affecting the mental health of patients, and even causing depression and suicide, and has become a public health problem that needs attention. The pathogenesis of acne is related to androgen-induced lipid secretion, abnormal keratinization of pilosebaceous duct, imbalance of skin surface microecology, and immune and inflammatory response. The current clinical treatment methods for acne mainly include external use and systemic drugs, such as oral antibiotics, hormone therapy, and retinoids. Although these methods have certain effects, the problems of antibiotic resistance, teratogenicity of isotretinoin, and side effects of long-term use of hormones are also the thorny problems in the clinical diagnosis and treatment of acne. Therefore, it is of great scientific and clinical value to develop new safe and targeted therapeutic targets or adjuvant drugs for the prevention and treatment of acne.

[0003] The intestinal microbiota is a vast microecosystem that plays an important role in maintaining intestinal-skin homeostasis. Intestinal flora can affect skin homeostasis by regulating immunity, metabolism, and neuroendocrine, and thus may affect the development of skin diseases. Intestinal flora is closely related to the occurrence and development of acne, mainly manifested as abnormal enrichment of pathogenic bacteria in the intestine and significant loss of protective flora. Studies have shown that after 300 acne patients were given probiotic tablets containing Lactobacillus acidophilus and Lactobacillus, 80% of the acne patients had a certain degree of clinical improvement. In a randomized clinical trial of oral probiotics for the treatment of acne vulgaris, it was found that oral probiotics (Lactobacillus rhamnosus) could significantly reduce the number of non-inflammatory acne lesions. In addition, studies have shown that Lactobacillus salivarius LS03 can be used as a "pseudo-antibiotic" for anti-inflammatory treatment of acne. However, the causal relationship between "missing intestinal microorganisms" and the occurrence and development of acne is not clear.

[0004] If specific missing protective bacteria for acne or drugs for the prevention and treatment of acne can be found. SUMMARY

[0005] The present application aims to provide a humanized probiotic strain and its application in the preparation of drugs for preventing or treating acne, in order to solve the problem of lack of clear and effective drugs for preventing and treating acne in the prior art.

[0006] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0007] A humanized Lactobacillus rhamnosus probiotic strain for preventing or treating acne, which is Lactobacillus rhamnosus, specifically named Lactobacillus rhamnosus GG SMUDH Yang02, and the abbreviation of the deposit unit is GDMCC, and the accession number is GDMCC No:65950, and the 16S rDNA sequence is shown as SEQ ID NO.1.

[0008] The application further provides an active metabolite of the humanized Lactobacillus rhamnosus probiotic strain for preventing or treating acne.

[0009] Further, the active metabolite comprises at least one of beta-galactosidase and soybean flavone.

[0010] The application further provides application of the humanized Lactobacillus rhamnosus probiotic strain and / or the active metabolite thereof in preparation of food, food additive, medicine for preventing or treating acne.

[0011] The application further provides a humanized probiotic composition comprising the humanized Lactobacillus rhamnosus probiotic strain Lactobacillus rhamnosus GG SMUDH Yang02 and / or the active metabolite thereof.

[0012] The application provides a humanized probiotic strain composition comprising Lactobacillus rhamnosus GG SMUDH Yang02 and Lacticaseibacillus paracasei (LP) YANG03, and the abbreviation of the deposit unit of the Lacticaseibacillus paracasei (LP) YANG03 is GDMCC, and the accession number is GDMCC No:65951, and the 16S rDNA sequence is shown as SEQ ID NO.2.

[0013] Further, the composition further comprises an active metabolite of the Lacticaseibacillus paracasei (LP) YANG03, and the active metabolite of the Lacticaseibacillus paracasei (LP) YANG03 comprises pantothenic acid.

[0014] The application further provides application of the humanized probiotic composition in preparation of food, food additive, medicine for preventing or treating acne.

[0015] The application also provides application of TFAP2A in preparation of a medicine for preventing or treating acne.

[0016] The application also provides application of a TFAP2A inhibitor in preparation of a medicine for preventing or treating acne.

[0017] The application has the advantages that the probiotics Lactobacillus rhamnosus GG SMUDH Yang02 bacteria and Lacticaseibacillus paracasei (LP) YANG03 bacteria, which are derived from the intestines of healthy people and are abnormally absent in the intestines of acne patients, and active metabolites thereof can specifically and efficiently prevent and treat acne. The Lactobacillus rhamnosus GG SMUDH Yang02 bacteria and the Lacticaseibacillus paracasei (LP) YANG03 bacteria can inhibit lipid synthesis of sebaceous glands and prevent acne progression in mice by secreting active metabolites. Metabolomics shows that the former can induce synthesis of Daidzein, and the latter can induce synthesis of pantothenic acid. The probiotics Lactobacillus rhamnosus GG SMUDH Yang02 bacteria and the Lacticaseibacillus paracasei (LP) YANG03 bacteria can fundamentally improve systemic inflammatory response and immune dysfunction by regulating intestinal microecological balance, and avoid problems of intestinal flora disorder and bacterial drug resistance caused by antibiotic treatment of acne. The probiotics Lactobacillus rhamnosus GG SMUDH Yang02 bacteria and the Lacticaseibacillus paracasei (LP) YANG03 bacteria can normally survive in the intestines of human bodies and have no toxic side effects. The application finds that TFAP2A can be used as a treatment target for acne and provides application of TFAP2A in preparation of a medicine for preventing and treating acne. BRIEF DESCRIPTION OF DRAWINGS

[0018] The accompanying drawings, which are included to provide a further understanding of the application and constitute a part of this application, illustrate embodiments of the application and together with the description serve to explain the application, and in which:

[0019] Figure 1 is a diagram of clinical cohort establishment and related research ideas for patients with moderate to severe acne as the first diagnosis;

[0020] Figure 2 is an experimental diagram for verifying that intestinal microecology is involved in the occurrence and development of acne;

[0021] Figure 3is the intestinal flora disorder of acne patients, and the statistical graph of Lactobacillus rhamnosus GG SMUDH Yang02 bacteria enrichment in the intestinal tract of healthy people;

[0022] Figure 4 is the experimental graph of verifying that Lactobacillus rhamnosus GG SMUDH Yang02 bacteria inhibit the occurrence and development of acne in mice;

[0023] Figure 5 is the experimental graph of verifying that the active metabolites released by Lactobacillus rhamnosus GG SMUDH Yang02 bacteria inhibit lipid synthesis in sebaceous gland cells;

[0024] Figure 6 is the experimental graph verifying that Lactobacillus rhamnosus GG SMUDH Yang02 bacteria can promote the synthesis of soy isoflavones (DAI) in vivo and in vitro;

[0025] Figure 7 is the experimental graph verifying that DAI can inhibit the formation of lipids in sebaceous gland cells and hinder the progression of acne in mice;

[0026] Figure 8 is the experimental graph verifying that DAI targets the expression of TFAP2A in sebaceous gland cells, and TFAP2A mediates the lipid synthesis inhibition effect of DAI;

[0027] Figure 9 is the experimental graph showing that Lacticaseibacillus paracasei (LP) YANG03 bacteria can significantly inhibit the number and size of acne lesions in mice;

[0028] Figure 10 is the experimental graph showing the influence of Lacticaseibacillus paracasei (LP) YANG03 bacteria on the expression of inflammatory genes, histopathology, sebaceous gland activity, and lesion thickness in mice with acne;

[0029] Figure 11 is the experimental graph of Lacticaseibacillus paracasei (LP) YANG03 bacteria inhibiting the occurrence and development of acne in mice, and the oil red O staining result;

[0030] Figures 12-13 is the analysis result graph of the composition of intestinal flora after different treatments;

[0031] Figure 14 is the experimental graph of non-target metabolomics sequencing in the feces of mice treated with Lacticaseibacillus paracasei (LP) YANG03 bacteria;

[0032] Figure 15 is the experimental graph of the appearance of the mouse acne model and skin lesions;

[0033] Figure 16 is the experimental graph of the height, number, size and scar score of the skin lesions of the mouse acne model in each experimental group;

[0034] Figure 17 is the experimental graph of the effect of pantothenic acid on the expression of pro-inflammatory cytokines in acne lesions;

[0035] Figure 18 is the skin sample tissue section of the mouse acne model stained with hematoxylin and eosin (H&E);

[0036] Figure 19 is Figure 18 the size of sebaceous glands, epithelial thickness, and sebaceous gland count histogram;

[0037] Figure 20 is the experimental graph of the treatment of mouse acne with Lactobacillus rhamnosus GG SMUDH Yang02 bacteria and Lacticaseibacillus paracasei (LP) YANG03 bacteria. DETAILED DESCRIPTION

[0038] The present application will be described in detail below in conjunction with the accompanying drawings and specific examples, which are illustrative of the present application and are used to explain the present application, but are not limiting to the present application.

[0039] The experimental methods in the following examples are all conventional methods unless otherwise specified, and the test materials used in the following examples are all commercially available unless otherwise specified. The standard for the healthy volunteers mentioned in the specification is: no bad habits, no use of probiotics and antibiotics and related products in the past 2 months, no history of tumors, infectious diseases. The requirements for acne patients are the same as those for healthy volunteers.

[0040] 1. Establishment of a clinical cohort of patients with moderate to severe acne at first visit

[0041] In order to analyze the composition and structure of the intestinal microbial community of acne patients as shown in Figure 1 , the present application establishes a clinical acne population cohort. A total of 29 acne patients and 20 healthy volunteers are included. The patients and healthy volunteers are enrolled and the baseline data are shown in Table 1:

[0042] Table 1. Baseline data and clinical characteristics of enrolled patients and healthy volunteers

[0043]

[0044] Fecal samples of enrolled patients, healthy volunteers were collected to extract fecal bacteria.

[0045] 2. Intestinal microecology participates in the occurrence and development of acne

[0046] In order to explore the potential relationship between intestinal flora and the occurrence and development of acne, the inventors constructed a mouse acne model induced by Propionibacterium acnes + 60% oleic acid. From the above enrolled healthy volunteers and acne patients, the fecal bacteria of healthy volunteers and acne patients were randomly selected, and the mouse acne model was orally perfused with PBS, healthy volunteer fecal bacteria and acne patient fecal bacteria (Fecal microbiota transplantation, FMT) after the mouse intestinal flora was cleared by antibiotics, as shown in Figure 2 A. The experimental results are as shown in Figure 2 B-2C. As can be seen from the size of the red circle, compared with the PBS solvent control group and the acne patient fecal bacteria transplantation group, the fecal bacteria from healthy volunteers can significantly inhibit the size (Progression) of the mouse acne-like skin lesions. At the same time, as shown in Figure 2 D, intestinal flora is significantly related to the occurrence of acne (Initiation). These results suggest that the fecal microbiota of healthy individuals can inhibit the development of mouse acne. Intestinal microecology participates in the evolution of acne.

[0047] 3. Intestinal flora of acne patients is disordered, and LGG bacteria and LP bacteria are enriched in the intestinal tract of healthy people

[0048] Through fecal sample metagenomic detection, it is found that, as shown in Figure 3 A, compared with the healthy volunteer control group, the intestinal microbial community composition of acne patients changes significantly, as shown in Figure 3 B, the intestinal microbial diversity of acne patients is significantly reduced. Further analysis found that Lactobacillus rhamnosus and Lacticaseibacillus paracasei were enriched in the intestinal tract of healthy people, but were abnormally absent in the intestinal tract of acne patients. It is suggested that the occurrence and development of acne is related to intestinal microecological disorder, and the absence of Lactobacillus rhamnosus and / or Lacticaseibacillus paracasei may be an important pathogenesis. As shown in Figure 3 C, the statistical graph of Lactobacillus rhamnosus enrichment in the intestinal tract of healthy people.

[0049] As shown in Figure 3To further explore the function and mechanism of the Lactobacillus rhamnosus involved in the evolution of acne, the Lactobacillus rhamnosus was independently isolated from the intestinal tract of a healthy volunteer, as shown in D. Whole genome detection of the bacteria found that the Lactobacillus rhamnosus was a new type of LGG bacteria from human intestinal tract, which was named Lactobacillus rhamnosus GG SMUDH Yang02 bacteria, and the 16S rDNA sequence thereof was shown as SEQ ID NO. 1.

[0050] SEQ ID NO. 1:

[0051]

[0052] Similarly, in order to further explore the function and mechanism of the paracasei involved in the evolution of acne, we independently isolated the paracasei from the intestinal tract of healthy volunteers. Whole genome detection found that:

[0053] The paracasei is a new type of human intestinal source LP bacteria, which is named Lacticaseibacillus paracasei (LP) YANG03 bacteria, and its 16S rDNA sequence is shown as SEQ ID NO. 2.

[0054] SEQ ID NO. 2:

[0055]

[0056] Different strains of different sources have different genomes and functions. Screening of disease-specific protective bacteria is crucial for elucidating the functions and potential mechanisms of the bacteria in disease prevention and treatment.

[0057] We preserved Lactobacillus rhamnosus GG SMUDH Yang02 bacteria, classified and named it as Lactobacillus rhamnosus, and preserved it at the Guangdong Microbial Culture Collection Center (GDMCC), located at No. 59, Building 5, Institute of Microbiology, Guangdong Academy of Sciences, 100, Martyrs Road, Yuexiu District, Guangzhou, Guangdong Province, China, on February 27, 2025, with the preservation number GDMCC No: 65950.

[0058] And we preserved Lacticaseibacillus paracasei (LP) YANG03 bacteria, classified and named it as Lacticaseibacillus paracasei, and preserved it at the Guangdong Microbial Culture Collection Center (GDMCC), located at No. 59, Building 5, Institute of Microbiology, Guangdong Academy of Sciences, 100, Martyrs Road, Yuexiu District, Guangzhou, Guangdong Province, China, on February 27, 2025, with the preservation number GDMCC No: 65951.

[0059] 4. LGG bacteria inhibit the development of acne in mice

[0060] As shown in Figure 4 A, to verify the relationship between Lactobacillus rhamnosus GG SMUDH Yang02 bacteria and the development of acne, we constructed a mouse acne model induced by Propionibacterium acnes + 60% oleic acid, and started feeding the mice with BHI (culture medium control), E. coli (bacterial control), and Lactobacillus rhamnosus GG SMUDH Yang02 bacteria orally every day for 2 weeks, then induced the mouse acne model by Propionibacterium acnes + 60% oleic acid. The experimental results are shown in Figure 4 B-4D, compared with BHI and E. coli controls, Lactobacillus rhamnosus GG SMUDH Yang02 bacteria can significantly inhibit the size of mouse acne lesions and reduce the expression of inflammatory genes such as TNF-ɑ, IL-6, and IL1-β. The histopathology and oil red O staining results are shown in Figure 4E shows that Lactobacillus rhamnosus GG SMUDH Yang02 bacteria can reduce subcutaneous abscesses and inflammatory cell infiltration and inhibit sebaceous secretion in mice compared with BHI and E. coli controls. It can be seen that Lactobacillus rhamnosus GG SMUDH Yang 02 bacteria are protective factors for the occurrence of acne, and targeted supplementation of the bacteria can resist the progression of acne.

[0061] 5. Lactobacillus rhamnosus GG SMUDH Yang02 bacteria release active metabolites to inhibit sebaceous gland cell lipid synthesis

[0062] The intestinal microecology regulates organs away from the intestine mainly through the active metabolites released by it. To verify whether the active metabolites metabolized by Lactobacillus rhamnosus GG SMUDH Yang02 bacteria can directly affect the number and function of sebaceous gland cells, we constructed a microbial active metabolite-sebaceous gland cell co-culture model. After co-culturing different concentrations (5%, 10%, 20% vol / vol) of Lactobacillus rhamnosus GG SMUDH Yang02 bacteria culture supernatant (Yang02.CM) with sebaceous gland cells SZ95, Nile red staining results as shown in Figure 5 A shows that Yang02.CM can significantly inhibit the lipid synthesis of SZ95 cells at concentrations of 10% and 20%, and BODIPY staining results as shown in Figure 5 B also confirm this conclusion. It can be seen that Lactobacillus rhamnosus GG SMUDH Yang02 bacteria release active metabolites that can directly target and inhibit the lipid synthesis function of sebaceous gland cells.

[0063] 6. Lactobacillus rhamnosus GG SMUDH Yang02 bacteria can promote the synthesis of Daidzein in vivo and in vitro

[0064] In order to identify the active metabolites that mediate Lactobacillus rhamnosus GG SMUDH Yang02 bacteria to target sebaceous gland cell lipid synthesis, we performed non-targeted metabolomics sequencing. The results are as shown in Figure 6 A-6B shows that the content of Daidzein (DAI for short) in Yang02.CM and the feces of mice treated with Lactobacillus rhamnosus GG SMUDH Yang02 bacteria is significantly increased. As shown in Figure 6As shown in C, Lactobacillus rhamnosus GG SMUDHYang02 bacteria and DAI were enriched in the intestinal tract of healthy people at the same time, but were abnormally absent in the intestinal tract of patients with acne vulgaris, which was consistent with the results of animal experiments. It can be seen that Lactobacillus rhamnosus GG SMUDHYang02 bacteria can induce the synthesis of DAI, and DAI may be an active metabolite that mediates the anti-acne effect of Lactobacillus rhamnosus GG SMUDHYang02 bacteria.

[0065] 7、DAI can inhibit lipid formation in sebocytes to resist the progression of acne-like skin lesions in mice

[0066] As Figure 7 As shown in A, the production of DAI is related to the decomposition of daidzin in food by intestinal microorganisms, and the decomposition process is mediated by β-galactosidase derived from microorganisms, and the gene coding frame of Lactobacillus rhamnosus GG bacteria can directly synthesize β-galactosidase; BHI, E. coli, Yang02.CM were added respectively in brain heart infusion medium, and BHI, E. coli, Lactobacillus rhamnosus GG SMUDHYang02 bacteria were used respectively to treat mouse feces, and targeted metabolomics detection showed that compared with BHI group and E. coli group, the content of daidzin in Yang02.CM group was significantly reduced, and the content of DAI in Yang02.CM group and mouse feces treated with Lactobacillus rhamnosus GG SMUDHYang02 bacteria was significantly increased, which was consistent with our guess. As Figure 7 As shown in B, among the two groups of mice with acne model respectively infused with solvent and Daidizein, the fluorescence quantitative analysis of oil red staining showed that the fluorescence of the Daidzein group was much less than that of the solvent group, indicating that the infusion of Daidzein can reduce the secretion of oil, and qPCR determination was performed on the skin of the two groups of mice, and it was found that IL-8, IL-1β, and lipid index SREBP1 were reduced in the Daidzein group, indicating that Daidzein can reduce acne inflammation and significantly inhibit the synthesis of sebocyte lipids to reduce sebum secretion. The results of constructing solvent and DAI treated mouse acne models are as follows Figure 7C-7F shows that each treatment group of mice acne lesions, DAI inhibition of mouse acne lesions score, each treatment group of mice lesion histopathology score results can be known, DAI gavage can significantly hinder the evolution of mouse acne. DAI is a kind of estrogen, estrogen can antagonize androgen-induced excessive secretion or accumulation of sebum, directly affect the occurrence and development of acne, which can indirectly explain the phenomenon of DAI targeting sebum secretion. The above results confirm that Lactobacillus rhamnosus GG SMUDH Yang02 bacteria decompose food source soybean glycosides by β-galactosidase to produce DAI, which can inhibit the lipid synthesis of sebocytes and prevent the progression of acne.

[0067] 8、DAI targets TFAP2A expression in sebocytes, and TFAP2A mediates the lipid synthesis inhibition effect of DAI

[0068] In order to explore the potential mechanism of DAI targeting the lipid synthesis function of sebocytes, we used small molecule receptor fishing technology (Lip-MS) to screen and fish the target points of DAI on sebocytes. The results are as follows Figure 8 A shows that TLR2 is a potential sebocyte binding target of DAI. Molecular docking is as follows Figure 8 B shows that the binding affinity of DAI to TLR2 receptor is-8.3 kcal / mol, and DAI can stably bind to TLR2 receptor of sebocytes. As shown in Figure 8 C, surface plasmon resonance technology (Surface plasmon resonance, SPR) further shows that 92 μM DAI captured by NTA chip occupies only half of the TLR2 binding site. These results suggest that DAI can stably bind to TLR2 receptor of sebocytes. In the process of acne vulgaris, P. acnes can bind to TLR2 receptor of sebocytes, promote the activation of downstream NF-kB pathway and induce inflammation and association reaction, and aggravate acne. The results above show that DAI can compete with P. acnes for binding to TLR2 receptor and inhibit its pro-inflammatory effect.

[0069] In order to find the key molecular events of DAI regulating the lipid synthesis function of sebocytes, as shown in Figure 8 D, we performed transcriptome sequencing on SZ95 sebocytes treated with DAI, and found that compared with the control group, DAI can significantly inhibit the Wnt receptor activity of SZ95 sebocytes. Among them, the transcription factor TFAP2 related to lipid synthesis also appears significant inhibition. Wnt signaling pathway can directly regulate lipid formation, and its induction of lipid droplet biogenesis depends on TFAP2 family transcription factors. As shown in Figure 8 E, q-PCR further shows that among the members of TFAP2 family, DAI can significantly inhibit the transcription of TFAP2A.Figure 8 As shown in Figure F, knockout of TFAP2A significantly reduced lipid synthesis in sebaceous gland cells and counteracted the anti-lipid synthesis efficacy of DAI. This indicates that DAI can inhibit TFAP2A transcription by binding to TLR2 and suppressing Wnt signaling, thereby interfering with lipid synthesis in sebaceous gland cells.

[0070] 9. LP bacteria inhibit the occurrence and development of acne in mice.

[0071] like Figure 9 As shown, to verify the relationship between *Lacticaseibacillus paracasei* (LP) YANG03 and the occurrence and development of acne, we constructed a mouse acne model induced by *Propionibacillus acnes* + 60% oleic acid. For the first two weeks, mice were orally fed BHI (culture medium control), *E. coli* (bacterial control), and *Lacticaseibacillus paracasei* (LP) YANG03 daily, respectively. Then, the mouse acne model was induced by *Propionibacillus acnes* + 60% oleic acid. The experimental results are shown below. Figures 9-11 As shown, compared with BHI and E. coli controls, *Lacticaseibacillus paracasei* (LP) YANG03 significantly inhibited the number and size of acne lesions in mice and reduced the expression of inflammatory genes such as TNF-α, IL-6, and IL-1β. Histopathological examination, sebaceous gland activity, lesion thickness, and Oil Red O staining results showed that, compared with BHI and E. coli controls, *Lacticaseibacillus paracasei* (LP) YANG03 reduced subcutaneous abscesses and inflammatory cell infiltration, and inhibited sebum secretion and keratinization in mice. Therefore, *Lacticaseibacillus paracasei* (LP) YANG03 is a protective factor against acne development, and targeted supplementation with this bacterium can resist acne progression.

[0072] 10. Lipoproteins of *Lacticaseibacillus paracasei* (LP) YANG03 regulate gut microbiota.

[0073] We performed metagenomic sequencing analysis on the composition of the gut microbiota after treatment, and the results are as follows: Figures 12-13 As shown, a significant increase in beneficial bacterial groups that contribute positively to gut health was found in the LP group, including:

[0074] The order Lachnospirales is known for its ability to produce short-chain fatty acids (SCFAs). SCFAs play a crucial role in reducing inflammation by inhibiting the NF-κB pathway. Lachnospirales are essential for maintaining a homeostatic inflammatory response in the gut; eubacteria, involved in SCFA production, contribute to anti-inflammatory effects and promote overall gut health.

[0075] Conversely, the LP group showed a reduction in the number of harmful bacteria from the Bacteroidetes phylum. These harmful bacteria are typically associated with various inflammatory diseases, and this reduction indicates a healthier gut microbiota structure in the LP group, suggesting a more balanced gut ecosystem. In stark contrast, the BHI and E. coli treatment groups exhibited poor gut microbiota. These groups displayed an imbalance, with a decrease in beneficial bacteria and an increase in potentially harmful strains, highlighting the negative impact of BHI and E. coli on gut health.

[0076] 11. *Lacticaseibacillus paracasei* (LP) YANG03 can promote the synthesis of vitamin B5 (pantothenic acid) both in vivo and in vitro.

[0077] To identify the bioactive metabolites that mediate lipid synthesis in sebaceous gland cells mediated by *Lacticaseibacillus paracasei* (LP) YANG03, we performed non-target metabolomics sequencing. The results are as follows: Figure 14 The results showed that pantothenic acid levels were significantly increased in the feces of mice treated with Lacticaseibacillus paracasei (LP)YANG03. Lacticaseibacillus paracasei (LP)YANG03 can induce pantothenic acid synthesis, and pantothenic acid may be an active metabolite mediating the anti-acne efficacy of Lacticaseibacillus paracasei (LP)YANG03.

[0078] 12. Pantothenic acid can inhibit lipid formation in sebaceous gland cells and thus resist the progression of acne-like lesions in mice.

[0079] Coenzyme A is crucial for the synthesis and metabolism of fatty acids, and pantothenic acid (vitamin B5) is a precursor to coenzyme A (CoA). Increased sebum production and proliferation of sebaceous glands are significant factors in the development and progression of acne. The bacterium *Lacticaseibacillus paracasei* (LP) YANG03, by producing pantothenic acid, may help regulate sebum secretion and reduce the severity of acne lesions. Figure 15As shown, the mice were orally fed with PBS or pantothenic acid respectively every day for one week before the construction of the P. acnes + 60% oleic acid-induced mouse acne model, and then the P. acnes + 60% oleic acid-induced mouse acne model was constructed, as shown in Figures 15-16 As shown, from the lesion height, number, size and scar score, it can be seen that pantothenic acid gavage can significantly hinder the evolution of mouse acne. As shown in Figure 17 As shown, pantothenic acid can reduce the expression of pro-inflammatory cytokines in acne lesions. As shown in Figure 18 As shown, the skin sample tissue sections were stained with hematoxylin and eosin (H&E), and the size of sebaceous glands and epithelial thickness under different treatment conditions were compared, and the statistical comparison results are as shown in Figure 19 As shown, the left graph shows the size of sebaceous glands in the PBS group and the pantothenic acid group (in μm²); the middle graph shows the lesion thickness in the control group and the pantothenic acid group (in μm²); and the right graph shows the sebaceous gland count in the PBS group and the pantothenic acid group. The above results confirm that Lacticaseibacillus paracasei (LP) YANG03 bacteria can inhibit sebaceous gland cell lipid synthesis and prevent acne progression by producing pantothenic acid.

[0080] 13. Lactobacillus rhamnosus GG SMUDH Yang02 bacteria and Lacticaseibacillus paracasei (LP) YANG03 bacteria in combination for treating mouse acne

[0081] In this experiment, four experimental groups of BHI, LGG, LP, and LGG+LP in combination were set up, and the mouse acne model was constructed according to the method described above. After 6 days, the appearance of the lesions was observed, and the lesion volume, height score, and overall score of each experimental group were recorded, as shown in Figure 20 As shown, the mouse acne treated with LGG+LP in combination was better than the mouse acne treated with LGG or LP alone in the above items.

[0082] The technical solutions provided by the embodiments of the present application are described in detail above, and specific examples are applied to describe the principles and implementation modes of the embodiments of the present application. The above description of the embodiments is only applicable to help understand the principles of the embodiments of the present application; at the same time, for those skilled in the art, according to the embodiments of the present application, the specific implementation modes and application scope will be changed, and the above description of the present application should not be understood as a limitation of the present application.

Claims

1. A humanized Lactobacillus rhamnosus probiotic strain that can prevent or treat acne, characterized in that, It is Lactobacillus rhamnosus, specifically named Lactobacillus rhamnosus GGSMUDH Yang02, with the depositary institution abbreviated as GDMCC and the accession number GDMCC No:65950. Its 16S rDNA sequence is shown in SEQ ID NO.

1.

2. An active metabolite of a humanized Lactobacillus rhamnosus probiotic strain as described in claim 1, which can prevent or treat acne.

3. The active metabolite of the humanized Lactobacillus rhamnosus probiotic strain for preventing or treating acne according to claim 2, characterized in that, The active metabolite includes at least one of β-galactosidase and soybean isoflavones.

4. The use of a humanized Lactobacillus rhamnosus probiotic strain and / or its active metabolites, as described in claim 1, in the preparation of food, food additives, or acne prevention or treatment drugs.

5. A humanized probiotic strain composition, characterized in that, Contains the humanized Lactobacillus rhamnosus GG SMUDH Yang02 strain as described in claim 1, with or without its active metabolites.

6. The humanized probiotic strain composition according to claim 5, characterized in that, The bacterium contained *Lacticaseibacillus paracasei* (LP)YANG03, whose depository unit is GDMCC and whose accession number is GDMCC No:65951, and whose 16S rDNA sequence is shown in SEQ ID NO.

2.

7. The humanized probiotic strain composition according to claim 5, characterized in that, The composition also includes an active metabolite of Lacticaseibacillus paracasei (LP)YANG03, which contains pantothenic acid.

8. The use of a humanized probiotic strain composition as described in any one of claims 5-7 in the preparation of food, food additives, or medicaments for the prevention or treatment of acne.

9. Application of TFAP2A in the preparation of drugs for the prevention or treatment of acne.

10. Application of TFAP2A inhibitors in the preparation of drugs for the prevention or treatment of acne.

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