Application of anthocyanin based on intestinal flora-intestinal barrier regulation in prevention and treatment of gestational diabetes mellitus

Abstract

The invention discloses application of anthocyanin based on intestinal flora-intestinal barrier regulation in prevention and treatment of gestational diabetes mellitus, and relates to the technical field of biological medicines, and the technical key points are as follows: the anthocyanin Cy-3-G can improve the intestinal barrier function and reduce inflammatory response by regulating intestinal flora, and can be used for preventing and treating gestational diabetes mellitus. Glucose homeostasis and insulin resistance of HFD mice can be effectively improved, complications of gestational diabetes mellitus are reduced, and health of mothers and infants is guaranteed.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to the application of anthocyanins based on the regulation of gut microbiota-intestinal barrier in the prevention and treatment of gestational diabetes mellitus. Background Art

[0002] Gestational diabetes refers to diabetes that first appears during pregnancy, usually occurring between 24 and 28 weeks of pregnancy. It is caused by hormonal changes in pregnant women leading to insulin resistance, thereby causing elevated blood glucose. Risk factors include overweight, family history, older age (such as over 35 years old), and pre-existing hyperglycemia before pregnancy, etc.

[0003] Although the existing technologies for the treatment and prevention of gestational diabetes are effective, there are still some drawbacks. First, the treatment relies on the self-management of pregnant women, including monitoring blood glucose, controlling diet, and exercise, which may be difficult for some pregnant women. Second, insulin treatment may cause side effects, such as hypoglycemic reactions, increasing the health risks of pregnant women and fetuses. In addition, strict diet control may affect the quality of life of pregnant women, and since each pregnant woman's condition is different, the treatment plan needs to be individualized and sometimes may not be able to effectively control blood glucose. Therefore, the existing technologies still face challenges such as poor compliance and potential side effects in the management of gestational diabetes.

[0004] Therefore, the present invention aims to provide the application of anthocyanins based on the regulation of gut microbiota-intestinal barrier in the prevention and treatment of gestational diabetes to solve the above problems. Summary of the Invention

[0005] The purpose of the present invention is to solve the above problems and provide the application of anthocyanins based on the regulation of gut microbiota-intestinal barrier in the prevention and treatment of gestational diabetes. The anthocyanin Cy-3-G of the present invention can effectively improve glucose homeostasis and insulin resistance in HFD mice, improve intestinal barrier function and reduce inflammatory responses by regulating gut microbiota, reduce the risks brought by gestational diabetes, and ensure the health of mothers and infants.

[0006] To achieve the above purpose, the technical solution of the present invention is as follows:

[0007] The present invention provides the application of anthocyanins based on the regulation of gut microbiota-intestinal barrier in the prevention and treatment of gestational diabetes. The anthocyanin is Cy-3-G, and the anthocyanin is a bilberry extract.

[0008] Anthocyanins can be used to prepare drugs for preventing and / or treating gestational diabetes. By regulating gut microbiota, anthocyanins can increase the abundance of Roseburia in gut microbiota, reduce the abundance of Lachnoclostridium, alleviate inflammatory responses, relieve glucose intolerance and insulin resistance, and intervene in the protective effect on intestinal barrier function at the protein and gene expression levels, thereby treating gestational diabetes.

[0009] Beneficial effects of this solution compared with the prior art:

[0010] The present invention discloses the application of anthocyanins based on the regulation of gut microbiota-intestinal barrier in the prevention and treatment of gestational diabetes, which relates to the field of biomedical technology. The technical key points are as follows: The anthocyanin Cy-3-G of the present invention can improve the intestinal barrier function and reduce the inflammatory response by regulating the gut microbiota, effectively improve glucose homeostasis and insulin resistance in HFD mice, reduce the occurrence of gestational diabetes complications, and ensure the health of mothers and infants. Brief Description of the Drawings

[0011] Figure 1 It is a schematic diagram of the GDM modeling process in the embodiment of the present invention;

[0012] Figure 2 It is a schematic diagram of glucose homeostasis in mice in the embodiment of the present invention; a is the pre-pregnancy OGTT of mice; b is the pre-pregnancy OGTT AUC of mice; c is the fasting insulin during pregnancy of mice; d is the OGTT during pregnancy of mice; e is the OGTT AUC during pregnancy of mice; f is the insulin resistance index during pregnancy of mice; *P<0.05 vs CON group, **P<0.01 vs CON group, ***P<0.001 vs CON group, #P<0.05 vs HFD group, ##P<0.01 vs HFD group;

[0013] Figure 3 It is a schematic diagram of the Alpha diversity analysis of the gut microbiota of mice in each group in the embodiment of the present invention; a is the ACE index; b is the Chao 1 index; c is the Shannon index; c is the Simpson index; ***P<0.001 vs CON group;

[0014] Figure 4 It is a schematic diagram of the Beta diversity analysis of the gut microbiota of mice in each group in the embodiment of the present invention; a is the PCoA analysis; b is the NMDS analysis; c is the Distances boxplot;

[0015] Figure 5 It is a schematic diagram of the community composition analysis of the gut microbiota of mice in each group in the embodiment of the present invention; a is the relative abundance at the phylum level; b is the relative abundance at the genus level;

[0016] Figure 6Schematic diagram of the species differences in the intestinal flora of mice in each group at the phylum level in the embodiments of the present invention. a is Actinobacteriota; b is Proteobacteria; c is Patescibacteria; d is Firmicutes; e is Bacteroidota; f is the F / B value, that is, Fimicutes / Bacteroidetes; *P<0.05;

[0017] Figure 7 Schematic diagram of the species differences in the intestinal flora of mice in each group at the genus level in the embodiments of the present invention. a is Lachnoclostridium; b is Roseburia;

[0018] Figure 8 Schematic diagram of the protein expressions of tight junction proteins ZO-1 and occludin in the colon tissues of mice in the embodiments of the present invention. Figure a is,...; Figure b is,...;

[0019] Figure 9 Schematic diagram of the levels of serum inflammatory factors in mice in the embodiments of the present invention. A is the LPS level; B is the IL-1β level; C is the TNF-α level; *P<0.05 vs CON group, **P<0.01 vs CON group, ***P<0.001 vs CON group, #P<0.05 vs HFD group, ##P<0.01 vs HFD group, P<0.001 vs HFD group. Detailed implementation manners

[0020] In order to enable those skilled in the art to better understand the solution of the present invention, the technical solution of the present invention will be further described in detail below in conjunction with the embodiments and drawings of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0021] It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other. The present invention will be described in detail below in conjunction with the embodiments.

[0022] Embodiment: Experimental method

[0023] 1. Experimental animals and grouping

[0024] 1.1. Experimental animals

[0025] Purchase 50 healthy specific pathogen-free (SPF) female C57BL / 6J mice (15±2 g) at 6 weeks of age and 25 SPF male C57BL / 6J mice (11 weeks old) (Jinan Pengyue Laboratory Animal Breeding Co., Ltd., animal certificate number: SCX(Lu)20190003). After checking that the appearance of the mice was normal, they were housed in an SPF animal room. Under a 12-hour light-dark cycle, the temperature (22±2 °C) and humidity (55±5%) were controlled, and sufficient food and water were provided. They were adaptively fed for 1 week. All animal experiments were conducted in accordance with the regulations of the Shandong Provincial Laboratory Animal Management Committee and were approved by the Laboratory Animal Ethics Committee of Jining Medical University (approval number: 2021-DW-ZR-027).

[0026] 1.2. Grouping and feeding of experimental animals

[0027] After one week of adaptive feeding of the 50 female mice, they were numbered (ear tags) in order of body weight and then divided into 5 groups (10 mice in each group) by the random number table method: normal control group (CON group), high-fat diet group (HFD group), high-fat diet + low-dose anthocyanin intervention group (LAC group), high-fat diet + medium-dose anthocyanin intervention group (MAC group), and high-fat diet + high-dose anthocyanin intervention group (HAC group). The normal control group was given a conventional low-fat diet (10% fat, Research Diets D12450B), and the high-fat diet group and anthocyanin intervention groups were given a high-fat diet (60% fat, Research Diets D12492). The energy intake efficiency of the 5 groups of mice was calculated according to the energy provided by the feed: energy intake efficiency (g / kcal) = [final body weight - initial body weight (g)] / simultaneous energy intake (kcal) × 100%, simultaneous energy intake (kcal) = total food intake (g) of each group of mice during the feeding period × energy provided per gram of feed, food intake = total feed given - remaining feed - spilled feed. In this experiment, the energy provided by feed D12450B in the CON group was 3.85 kcal / g, and the energy provided by high-fat feed D12492 in the HFD group and each intervention group was 5.24 kcal / g.

[0028] 2. Blueberry extract addition dose

[0029] Among the anthocyanins in food, cyanidin-3-O-glucoside (Gyanidin-3-O-glucoside, Cy-3-G) is the most widely distributed. Therefore, the total anthocyanins in food can generally be converted to Cy-3-G for quantification. In this experiment, the anthocyanins used were bilberry extract (Mirtoselect), with a Cy-3-G concentration of 41.6% and a production batch number of 19S0155900. The intervention method was feed addition, and the intervention doses were set based on the results of the preliminary experiment as follows: LAC group: 0.01% (w,w), MAC group: 0.05% (w,w), HAC group: 0.1% (w,w).

[0030] 3. Establishment of gestational diabetes mellitus animal model

[0031] After 1 week of adaptive feeding, the model was established. The CON group was given a conventional low-fat diet (Research Diets D12450B), and the HFD group was given a high-fat diet (Research Diets D12492), with the fat energy ratios being 10% and 60% respectively. The modeling period included 6 weeks before pregnancy and 16 days of pregnancy. Fasting blood glucose (FBG) was measured before the start of modeling and 6 weeks after intervention, and mice with abnormal blood glucose (FBG≥7 mmol / L) were excluded. And an oral glucose tolerance test (OGTT) was performed on the mice 6 weeks after intervention. Then, female C57BL / 6J mice were caged with male C57BL / 6J mice at a ratio of 2:1. At 7:00 in the morning the next day, check whether the female mice have vaginal plugs. Those with vaginal plugs are considered to have successful mating. Those without vaginal plugs are examined by vaginal smear. If sperm can be seen under the microscope after smearing, it is considered successful mating. The successfully mated female mice and male mice were separated and raised, and it was recorded as the 0.5th day of pregnancy (G0.5d). On the 15.5th day of pregnancy (G15.5d), after fasting for 12 hours, FBG was measured and OGTT was performed. According to the significant increase in FBG, insulin level and insulin resistance index, it was suggested that the pregnant mice fed with a high-fat diet met the characteristics of pathological insulin resistance in gestational diabetes patients, and the GDM mouse model was successfully established. On the 16.5th day of pregnancy (G16.5d), after fasting for 12 hours, the mice were weighed, sacrificed, and samples were taken. During the GDM modeling period, the experimental female mice were weighed twice a week and the food intake of the mice was recorded daily. The feeding and intervention methods of each group of mice remained unchanged. The specific process of GDM modeling is shown in Figure 1 。

[0032] 4. Oral glucose tolerance test

[0033] (1) Measurement of blood glucose level: OGTT experiments were conducted 6 weeks after the intervention of mice and at 15.5 days of pregnancy. The mice were fasted for 12 hours at 8 pm the day before the experiment until 8 am the next day. During the fasting period, the mice were allowed to drink normal water. Prepare 20% glucose solution in advance and place it at room temperature overnight. At 8 am, take the mice out of the cage, weigh each mouse and record it. Use a sterilized scissors to cut about 1 - 2 mm from the end of the mouse's tail. Gently squeeze the mouse's tail along the tail vein to make a drop of blood accumulate. After wiping off the first drop of blood with a medical cotton swab, use a blood glucose meter and blood glucose test strip to measure the blood glucose level of the second drop of blood. The measured value is defaulted to the blood glucose level at 0 minute (G0), that is, FBG. After the mice are in a stable state, connect a 1 ml syringe to a gastric gavage needle, calculate and draw a certain volume of 20% glucose solution (2 g / kg body weight) according to the body weight, and conduct gastric gavage on the mice. The operation interval for each mouse is about 30 seconds. Start timing from the completion of gastric gavage of the first mouse. Measure the blood glucose levels at the tail tip of the mouse (G30, G60, G90, G120) at 30 minutes, 60 minutes, 90 minutes, and 120 minutes respectively using a blood glucose meter. Operate gently to avoid stress hyperglycemia in mice. After the operation, disinfect the tail tip with iodophor.

[0034] (2) Area under the curve (AUC) of blood glucose was calculated according to the approximate trapezoidal area formula. The formula is: AUC = 30×[(G0 + G120) / 2 + G30 + G60 + G90], and the unit is mmol / L*120min.

[0035] 5. Collection of animal tissue samples

[0036] At 16.5 days of pregnancy in mice, weigh the pregnant mice, fast them for 12 hours, take blood by removing the eyeballs of the mice. After the blood collection is completed, let it stand at room temperature for 2 hours. After the blood coagulates and the blood clot contracts, centrifuge at 3500 revolutions per minute for 10 minutes at 4°C. Gently suck the supernatant with a pipette tip and transfer it to a clean centrifuge tube. Store it in an -80°C refrigerator in aliquots of 150 ul per tube for subsequent experiments.

[0037] Decapitate the mice after blood collection and quickly dissect them on the operating table. Collect the liver, pancreas, colon, perirenal adipose tissue, colonic contents, cecal contents, and feces of the pregnant mice. Accurately weigh the weight of each tissue and record it. Finally, store it in an -80°C refrigerator for subsequent detection and analysis.

[0038] 6. Determination of serum fasting insulin (FINS), IL-1β, TNF-α, and LPS concentrations

[0039] The serum insulin level of mice was detected using an enzyme-linked immunosorbent assay (ELISA) kit. The specific steps were carried out according to the instructions. The insulin resistance index (HOMA-IR) was calculated to evaluate insulin sensitivity. The HOMA-IR calculation formula was:

[0040]

[0041] Serum IL-1β, TNF-α and LPS concentrations were also measured according to the instructions of the corresponding kits.

[0042] 7.16S rRNA high-throughput sequencing to detect the composition and analysis of intestinal flora in mouse colon contents

[0043] (1) DNA extraction and quality inspection: Genomic DNA was extracted using an environmental sample DNA extraction kit (OMEGA). The integrity of the extracted genomic DNA was tested using 1% agarose gel electrophoresis, and the DNA concentration was quantified using the Qubitpicogreen fluorescence quantitative system.

[0044] (2) Target region amplification and product purification: According to the specified sequencing region, specific primers with barcode sequences are synthesized. Low cycle amplification is used to ensure that the number of cycles of sample amplification is consistent to ensure the accuracy and consistency of data analysis. The amplified product is detected by 2% agarose gel electrophoresis, recovered and purified using magnetic beads, and used for quantitative library construction after passing the inspection;

[0045] (3) Fluorescence quantification: The concentration of the purified PCR products was quantified using the Qubit picogreen fluorescence quantification system, and the products were mixed in corresponding proportions according to the sequencing amount required for each sample to construct a sequencing library;

[0046] (4) Sequencing: The constructed library was quantified using Qubit, and the linker efficiency was detected using qPCR. After the actual concentration was calculated based on the obtained efficiency, the library was diluted to a certain concentration according to the requirements of the machine and sequenced using NovaSeq.

[0047] 8. Immunofluorescence of Colon Tissue

[0048] (1) Dewaxing of paraffin sections: Place the sections in environmentally friendly dewaxing solution I for 10 min, environmentally friendly dewaxing solution II for 10 min, environmentally friendly dewaxing solution III for 10 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, and then wash with distilled water.

[0049] (2) Antigen repair: During the repair process, the buffer should be prevented from evaporating excessively and the slides should not be dried. After the repair is completed, cool naturally. Place the slides in PBS (PH7.4) and wash them on a decolorizing shaker three times, 5 minutes each time.

[0050] (3) Circle serum blocking: After gently shaking off the excess liquid from the sections, use a histochemical pen to draw a circle around the tissue, and add BSA, then incubate for 30 minutes for blocking.

[0051] (4) Add the mixed reagent of the first and second primary antibodies: Mix two primary antibodies from different sources, add the prepared primary antibody, and place the sections flat in a humid chamber and incubate overnight at 4°C.

[0052] (5) Add the secondary antibody: Place the slides in PBS (pH 7.4) and wash on a shaker for 3 times, 5 minutes each time. Add the corresponding secondary antibody and incubate at room temperature in the dark for 50 minutes.

[0053] (6) DAPI counterstain the cell nuclei: Place the slides in PBS (pH 7.4) and wash on a shaker for 3 times, 5 minutes each time. Add DAPI staining solution and incubate at room temperature in the dark for 10 minutes.

[0054] (7) Quench the autofluorescence of the tissue: Place the slides in PBS (pH 7.4) and wash on a shaker for 3 times, 5 minutes each time. Add the autofluorescence quenching agent B solution for 5 minutes and then rinse with running water for 10 minutes.

[0055] (8) Mount the slides: Mount the slides with an anti-fluorescence quenching mounting medium.

[0056] (9) Acquire images: The excitation wavelength of DAPI is 330 - 380 nm, and the emission wavelength is 420 nm; the excitation wavelength of 488 is 465 - 495 nm, and the emission wavelength is 515 - 555 nm; the excitation wavelength of CY3 is 510 - 560 nm, and the emission wavelength is 590 nm; the excitation wavelength of CY5 is 608 - 648 nm, and the emission wavelength is 672 - 712 nm.

[0057] Results:

[0058] 1. Effects of anthocyanins on glucose homeostasis in mice

[0059] After 50 female mice were raised for 6 weeks and then caged with male mice for 1 week, a total of 36 mice (6 mice in the CON group, 6 mice in the HFD group, 6 mice in the LAC group, 9 mice in the MAC group, and 9 mice in the HAC group) became pregnant after caging. To successfully establish a mouse GDM model, the FBG of 5 groups of female mice was detected before caging, and the mice with FBG ≥ 7.0 mmol / L were excluded. And the OGTT before caging was detected, and the AUC was calculated. There were no significant differences among the 5 groups ( Figure 2 a, b, P > 0.05). During pregnancy, compared with the CON group, the OGTT AUC, insulin level, and insulin resistance index in the HFD group were all significantly increased ( Figure 2d, e, c, f, *P < 0.05, **P < 0.01, ***P < 0.001). Significantly impaired glucose tolerance and insulin resistance were observed in the HFD group, indicating the successful establishment of a high-fat diet-induced GDM model in mice. Compared with the HFD group, the OGTTAUC, fasting insulin, and HOMA-IR of the MAC group and the HAC group mice were significantly decreased, indicating that the addition of high-dose blueberry extract to the diet could significantly alleviate high-fat diet-induced glucose intolerance and insulin resistance. Figure 2 c, e, f, #P < 0.05, ##P < 0.01).

[0060] 2. Effects of anthocyanins on the intestinal flora of mice

[0061] 2.1. Alpha diversity analysis

[0062] As Figure 3 shown, compared with the CON group, the chao index and ACE index of the HFD group were significantly increased (P < 0.001), indicating that the intestinal flora colony richness of the HFD group mice was significantly higher than that of the CON group. There was no significant difference in the shannon of the intestinal flora in the colon contents of the five groups of mice.

[0063] 2.2. Beta diversity analysis

[0064] The method of principal coordinates analysis (PCoA) can show the difference size between the samples of each group. The dots of each color represent the mouse samples within different groups. If the distance between two samples is close, it means that the species composition of the intestinal flora of these two mice is more similar. As Figure 4 a, the HFD group and the LAC group almost overlapped, indicating that the difference in species diversity between the two groups was small, while compared with the other three groups (CON group, MAC group, and HAC group), the difference between the groups was obvious. Nonmetric Multidimensional Scaling (NMDS) can represent the difference size of species diversity based on evolutionary relationships or quantitative distance matrices. Generally, when stress < 0.2, the NMDS graph has a certain explanatory significance, and when stress < 0.1, it is considered a good ranking. As Figure 4 b, except that the HFD group and the LAC group were close in distance, the distances between the other groups were far, and stress = 0.062, indicating that the grouping was meaningful. Except for the small difference in species diversity between the HFD group and the LAC group, the species diversity of the intestinal flora of the mice in the other groups was significantly different. The distance between the sample groups was statistically analyzed at the OTU level. At Figure 4In c, the boxes corresponding to "Between" represent the distance values of the between-group differences, and the remaining boxes represent the within-group difference distance values, further demonstrating with data that the species diversity between the intestinal microbiota of the 5 groups of samples is large while the within-group species diversity is small (R = 0.93, P = 0.001).

[0065] 2.3. Community composition analysis

[0066] To further study the effect of blueberry extract on the intestinal microbiota distribution of GDM mice, the relative abundances of species in each group were analyzed at the phylum and genus levels. As Figure 5 shown in the a bar chart, the phyla with relatively high rankings in terms of species relative abundance at the phylum level are: Firmicutes, Bacteroidetes, Actinobacteria, Campylobacterota, Desulfobacterota, Verrucomicrobiota, Proteobacteria, etc. Figure 5 As shown in the b bar chart, the relative abundances of species in the intestinal microbiota of mice were also statistically analyzed at the genus level. The genera with relatively high rankings in the relative abundances of the intestinal microbiota of each group of mice are not completely the same. Generally, the genera with relatively high abundances are: unclassified_f__Muribaculaceae, Faecalibaculu, unclassified_f__Lachnospiraceae, Bacteroides, etc.

[0067] 2.4. Species difference analysis

[0068] Based on the dominant populations with relatively high rankings in the relative abundances of the intestinal microbiota of each group of mice at the phylum and genus levels, species difference analyses were performed at the phylum and genus levels respectively. As Figure 6 shown, at the phylum level, compared with the CON group, the abundances of Actinobacteriota, Proteobacteria, and Patescibacteria in the HFD group decreased significantly (P < 0.05). Compared with the HFD group, the abundances of Actinobacteriota in the MAC group and the HAC group increased significantly (P < 0.05). There were no significant differences in the abundances of Firmicutes, Bacteroidota, and the ratio of their abundances (F / B value) among the 5 groups of samples.

[0069] Figure 7Species difference analysis at the genus level. Lachnoclostridium showed an upward trend in the HFD group and a downward trend in the HAC group; Roseburia showed a downward trend in the HFD group and upward trends in the MAC and HAC groups (P<0.05).

[0070] 3. Effect of anthocyanins on the expression of tight junction proteins in the mouse colon

[0071] As shown in the figure, tight junction proteins were expressed on the luminal surface of colonic villi and the surface of crypt epithelium ( Figure 8 A). The fluorescence intensity of ZO-1 in the HFD group was lower than that in the CON, LAC, MAC, and HAC groups, but did not reach statistical significance (P>0.05, Figure 8 B). The fluorescence intensity of Occludin in the HFD group was significantly lower than that in the CON group and significantly higher than that in the HFD group in all three intervention groups (P<0.05, Figure 8 C). The mRNA levels of the two tight junction proteins were significantly lower in the HFD group than in the CON group (P<0.05, Figure 8 D), and significantly higher in the MAC group than in the HFD group (P<0.01, Figure 8 E). The results indicate the protective effect of anthocyanin intervention on intestinal barrier function at the protein and gene expression levels.

[0072] 4. Effect of anthocyanins on the inflammatory level of mice

[0073] Compared with the CON group, the serum LPS, IL-1β, and TNF-α of mice in the HFD group were significantly increased; compared with the HFD group, the serum LPS concentration decreased in the LAC group, the serum LPS and IL-1β concentrations decreased in the MAC group, and the serum LPS, IL-1β, and TNF-α concentrations all showed a downward trend in the HAC group ( Figure 9 a-c). The results show that the intervention of bilberry extract can effectively improve the inflammatory response of mice caused by high-fat diet.

[0074] The above specific embodiments are only explanations of the present invention, and they are not limitations of the present invention. Those skilled in the art can make modifications without creative contributions to the embodiments according to needs after reading this specification, but as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. The application of anthocyanins in the prevention and treatment of gestational diabetes based on intestinal flora-intestinal barrier regulation is characterized by: The anthocyanin is Cy-3-G, and the anthocyanin is bilberry extract.

2. The use of anthocyanins based on intestinal flora-intestinal barrier regulation in the prevention and treatment of gestational diabetes as claimed in claim 1, characterized in that: The anthocyanin can be used to prepare medicine for preventing and / or treating gestational diabetes.

3. The use of anthocyanins based on intestinal flora-intestinal barrier regulation in the prevention and treatment of gestational diabetes as claimed in claim 2, characterized in that: The anthocyanins treat gestational diabetes by regulating intestinal flora, increasing the relative abundance of beneficial bacteria, reducing the relative abundance of harmful bacteria, alleviating inflammatory responses, alleviating glucose intolerance and insulin resistance, and intervening in the protection of intestinal barrier function at the protein and gene expression levels.

4. The use of anthocyanins based on intestinal flora-intestinal barrier regulation in the prevention and treatment of gestational diabetes as claimed in claim 3, characterized in that: The anthocyanins can increase the abundance of Roseburia and reduce the abundance of Lachnoclostridium in the intestinal flora.

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