阅读说明：本技术 亚麻籽油在制备改善动脉粥样硬化症肠道菌群的药物中的应用 (Application of linseed oil in preparation of medicine for improving atherosclerosis intestinal flora ) 是由 王浩 李一唯 张晓霞 贾绍斌 汪婷 柳媛媛 于 2021-08-19 设计创作，主要内容包括：本发明提供了亚麻籽油在制备改善动脉粥样硬化症肠道菌群的药物中的应用,属于生物医药技术领域。亚麻籽油具备调节动脉粥样硬化症肠道菌群的作用。小鼠动物模型试验显示,亚麻籽油能够改变动脉粥样硬化症肠道菌群,并改变肠道菌群代谢产物短链脂肪酸水平和代谢产物胆汁酸水平,同时,动脉粥样硬化症得到减轻,说明亚麻籽油能够通过调节肠道菌群来改善脉粥样硬化症。因此,可将亚麻籽油用作制备改善动脉粥样硬化症肠道菌群的药物,以缓解动脉粥样硬化症病程的发展。(The invention provides an application of linseed oil in preparation of a medicine for improving atherosclerosis intestinal flora, and belongs to the technical field of biological medicines. The linseed oil has the function of regulating intestinal flora of atherosclerosis. Animal model experiments of mice show that the linseed oil can change the intestinal flora of atherosclerosis, change the short-chain fatty acid level of metabolites of the intestinal flora and the bile acid level of the metabolites, and simultaneously relieve the atherosclerosis, which indicates that the linseed oil can improve the atherosclerosis by regulating the intestinal flora. Therefore, the linseed oil can be used for preparing a medicament for improving the intestinal flora of atherosclerosis so as to relieve the development of the disease course of the atherosclerosis.)

1. Application of linseed oil in preparation of medicines for improving intestinal flora of atherosclerosis is provided.

2. Use of linseed oil according to claim 1 in the preparation of a medicament for improving the intestinal flora of atherosclerosis characterized in that: the linseed oil can relieve atherosclerosis by regulating intestinal flora and changing the short-chain fatty acid level of metabolites and the bile acid level of metabolites of the intestinal flora.

3. Use of linseed oil according to claim 2 in the manufacture of a medicament for improving the intestinal flora of atherosclerosis characterized in that: the intestinal flora comprises Intestiminonas, Bilophila, Anaerotruncus, Oscillbacter, Negatibacter, Lachnocrostidium and Enterorhabdus, and the linseed oil can reduce the flora.

4. Use of linseed oil according to claim 2 in the manufacture of a medicament for improving the intestinal flora of atherosclerosis characterized in that: the linseed oil can increase the content of acetic acid, propionic acid, isovaleric acid, isobutyric acid and valeric acid in intestinal flora metabolite short-chain fatty acid; the linseed oil can reduce the content of alloLCA (allolithocholic acid), isoLCA (isocholic acid), 7-ketoLCA (7-ketolithocholic acid), beta-UDCA (3 beta-ursodeoxycholic acid), CDCA (chenodeoxycholic acid) and HDCA (hyodeoxycholic acid) in intestinal flora metabolite bile acid, and increase the content of LCA (lithocholic acid), ACA (allocholic acid), GCA (glycocholic acid) and TCA (taurocholic acid).

5. A medicine for improving atherosclerosis intestinal flora is characterized in that: the medicine is mainly prepared from effective dose of linseed oil and pharmaceutically acceptable auxiliary materials according to a conventional preparation method.

6. The medicament for improving the intestinal flora of atherosclerosis according to claim 5, wherein: the dosage form of the medicine comprises tablets, capsules, lotion, granules or oral preparation.

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of linseed oil in preparation of a medicine for improving atherosclerosis intestinal flora.

Background

Atherosclerosis (AS) is a major cause of cardiovascular disease, a chronic inflammatory disease with lipid disorders and plaque instability characterized by excessive deposition of cholesterol in the arterial wall. Atherosclerosis is also a systemic disease, and if atherosclerotic lesions develop, it means that the same lesions may develop in blood vessels elsewhere. At present, the pathogenesis of atherosclerosis is quite complex, and the specific etiology has no definite conclusion.

The intestinal flora is the microbial flora inhabiting in human intestinal tracts, and the number of bacteria is about 40 trillion. The intestinal microorganisms form an intestinal mucosa barrier and play an important role in maintaining the normal physiological functions of the organism, including regulating the absorption and metabolism of nutrients, avoiding lipid metabolism disorder, regulating the immune system and the like. When the internal and external environments of the body change, the intestinal flora is unbalanced, thereby causing various diseases, such as gastrointestinal diseases, metabolic diseases, nervous system diseases, cardiovascular diseases and the like, and causing serious harm to the body. Research has proved that atherosclerosis is closely related to intestinal flora, imbalance of the intestinal flora can cause metabolic imbalance and inflammatory reaction, so that plaque is formed and ruptured, and imbalance of the intestinal flora can change the level of flora metabolites, so that the pathogenesis of atherosclerosis is influenced.

Flaxseed Oil (FO), one of the major edible oils worldwide, is a major component extracted from Flaxseed, and is also an important plant source of α -linolenic acid (α -linolenic acid, ALA, octadeca 9, 12, 15-trienoic acid). Alpha-linolenic acid has been shown to improve blood lipids, lower blood pressure and reduce inflammatory factors. The content of Polyunsaturated fatty acids (PUF As) in linseed oil is As high As 77.51% -92.39%, and the linseed oil is rich in alpha-linolenic acid (53% -65%). At present, linseed oil is mainly used for improving atherosclerosis through anti-inflammation, anti-oxidative stress, lipid metabolism regulation, vascular endothelial cell function promotion and the like. There is no report that linseed oil improves atherosclerosis by regulating intestinal flora.

Disclosure of Invention

In view of the above, the present invention aims to provide an application of linseed oil in preparing a medicament for improving the intestinal flora of atherosclerosis, and to expand the application range of linseed oil.

The technical scheme adopted by the invention for solving the technical problems is as follows:

the invention provides an application of linseed oil in preparing a medicine for improving atherosclerosis intestinal flora.

Preferably, the linseed oil is used for alleviating atherosclerosis by regulating intestinal flora and changing the short-chain fatty acid level of metabolites and the bile acid level of metabolites of the intestinal flora.

Preferably, the gut flora comprises intestinomonas, Bilophila (cholephilus), anaerobot runcus (anaerobiosharomyces), Oscillibacter, negavibacillus, lachnoclostrium and Enterorhabdus, and the linseed oil is capable of down-regulating the flora.

Preferably, the linseed oil can increase the content of acetic acid, propionic acid, isovaleric acid, isobutyric acid and valeric acid in short-chain fatty acids which are metabolites of intestinal flora; the linseed oil can reduce the content of alloLCA (allolithocholic acid), isoLCA (isocholic acid), 7-ketoLCA (7-ketolithocholic acid), beta-UDCA (3 beta-ursodeoxycholic acid), CDCA (chenodeoxycholic acid) and HDCA (hyodeoxycholic acid) in intestinal flora metabolite bile acid, and increase the content of LCA (lithocholic acid), ACA (allocholic acid), GCA (glycocholic acid) and TCA (taurocholic acid).

A medicine for improving intestinal flora of atherosclerosis is mainly prepared from effective amount of linseed oil and pharmaceutically acceptable auxiliary materials according to a conventional preparation method.

Preferably, the dosage form of the medicament comprises tablets, capsules, lotion, granules or oral preparations.

According to the technical scheme, the beneficial effects of the invention are as follows: the invention provides an application of linseed oil in preparing a medicine for improving atherosclerosis intestinal flora. The linseed oil has the function of regulating intestinal flora of atherosclerosis. Rigorous scientific experiments prove that the linseed oil can regulate the intestinal flora of the atherosclerosis, change the short-chain fatty acid level of metabolites of the intestinal flora and the bile acid level of the metabolites, and relieve the atherosclerosis, so that the linseed oil can improve the atherosclerosis by regulating the intestinal flora. Therefore, the linseed oil can be used for preparing a medicament for improving the intestinal flora of atherosclerosis so as to relieve the development of the disease course of the atherosclerosis.

Drawings

FIG. 1 shows the histopathological staining results of the mice of each group in the experimental examples, note: a scale: 500 μm.

Fig. 2 is a graph of alpha diversity analysis of the effect of linseed oil on the intestinal flora of atherosclerotic mice.

FIG. 3 is a graph of beta-diversity analysis of the effect of linseed oil on the intestinal flora of mice with atherosclerosis.

FIG. 4 is a graph of analysis of the phylum level of the effect of linseed oil on the composition of the intestinal flora of atherosclerotic mice.

FIG. 5 is a genus level analysis chart of the effect of linseed oil on the composition of intestinal flora in mice with atherosclerosis.

FIG. 6 is a short chain fatty acid peak plot and a differential short chain fatty acid clustering heatmap.

FIG. 7 shows the effect of linseed oil on short chain fatty acids, metabolites of the intestinal flora of atherosclerotic mice.

Fig. 8 is a bile acid peak plot and a differential bile acid clustering heatmap.

Fig. 9 shows the effect of linseed oil on bile acid, a metabolite of the intestinal flora of mice with atherosclerosis.

Detailed Description

The technical scheme and the technical effect of the invention are further elaborated in the following by combining the drawings of the invention.

The invention provides an application of linseed oil in preparing a medicine for improving atherosclerosis intestinal flora.

Furthermore, the linseed oil can be used for relieving atherosclerosis by regulating intestinal flora and changing the short-chain fatty acid level of metabolites and the bile acid level of metabolites of the intestinal flora.

Further, the intestinal flora includes intestinomonas, Bilophila (cholephilus), Anae rotillus (anaerobic clavulans), Oscillibacter, negavibacillus, lachnoclostrium and Enterorhabdus, and the linseed oil can down-regulate the flora.

Further, the linseed oil can increase the content of acetic acid, propionic acid, isovaleric acid, isobutyric acid and valeric acid in intestinal flora metabolite short-chain fatty acids; the linseed oil can reduce the content of alloLCA (allolithocholic acid), isoLCA (isocholic acid), 7-ketoLCA (7-ketolithocholic acid), beta-UDCA (3 beta-ursodeoxycholic acid), CDCA (chenodeoxycholic acid) and HDCA (hyodeoxycholic acid) in intestinal flora metabolite bile acid, and increase the content of LCA (lithocholic acid), ACA (allocholic acid), GCA (glycocholic acid) and TCA (taurocholic acid).

A medicine for improving intestinal flora of atherosclerosis is mainly prepared from effective amount of linseed oil and pharmaceutically acceptable auxiliary materials according to a conventional preparation method.

Further, the dosage form of the medicine comprises tablets, capsules, lotion, granules or oral preparations.

In order to confirm that linseed oil has the above-mentioned new functions, the following tests were performed:

the linseed oil in the test is extracted by an oil extraction instrument according to a conventional operation mode, the oil extraction instrument in the test adopts an Auckoma household oil press with the model AZ-B301, and the content of alpha-linolenic acid is 59.58% +/-2.47.

Test example 1 Effect of linseed oil on mice with Atherosclerosis

30 male ApoE^-/-Mice (8 weeks old) were purchased from Beijing Wittingle laboratory animal technology Ltd (license number: SCXK (Jing) 2016-. After the mice are adaptively raised for 1 week, the weight of the mice is adjustedThe machine is divided into 3 groups (10/group), which are respectively: blank control group (CON), atherosclerosis model group (MOD) and linseed oil pretreatment group (MOD/FO). Wherein, the placebo group (CON) mice were given a normal diet, the atherosclerosis model group (MOD) mice were fed with a high fat diet containing 1.25% (w/w) cholesterol, and the linseed oil dry pre-group (MOD/FO) mice were fed with a high fat diet containing 10% (w/w) linseed oil (100 g of FO instead of isocaloric cocoa butter per kg of feed) and 1.25% (w/w) cholesterol. The diet dose was the same for each group of mice, and the mice were continuously kept for 12 weeks. Mice of a blank control group (CON), an atherosclerosis model group (MOD) and a linseed oil pretreatment group (MOD/FO) after being bred for 12 weeks are randomly picked respectively, the mice are anesthetized, dissected and placed under a body type microscope, and aorta and aortic sinus are separated for pathological staining. Aorta was stained with gross oil red O; embedding the aortic sinus specimen in a tissue freezing culture medium, continuously slicing into slices with the thickness of 8mm at the temperature of-20 ℃, and respectively adopting HE staining, oil red O staining and masson staining; the stained tissue sections were placed under an optical microscope to observe each group of pathological conditions and quantitatively analyzed using Image J software.

As a result: referring to fig. 1, in fig. 1, a is a graph of aorta gross oil red O staining results, B is a graph of quantitative analysis of plaques in aorta, C is a graph of aortic sinus HE staining, masson staining and oil red O staining results of each group, D is quantitative analysis of aortic sinus plaque fibrosis, and E is quantitative analysis of aortic sinus lipid plaque. The results show that the atherosclerosis model group (MOD) has significantly increased aortic and aortic sinus lipid plaques compared with the blank control group (CON), and the differences are statistically significant (P < 0.05). In the linseed oil intervention group (MOD/FO), aortic and aortic sinus lipid plaques were reduced, while the vessel wall at the aortic root was significantly thickened, the lumen was narrowed, and the difference was statistically significant (P < 0.05). The masson staining result shows that the atherosclerosis model group (MOD) has obviously increased arterial vascular fibrosis level (P < 0.05); the level of vascular fibrosis is reduced after linseed oil is dried. Meanwhile, conventional pathological HE staining indicates that pathological damage of arterial vessels of an atherosclerosis model group (MOD) is serious, linseed oil is dry, and pathological damage of the aorta is partially reversed. The above results all confirm that long-term intervention of linseed oil can significantly slow down lipid deposition of atherosclerotic vascular plaques, thereby contributing to improvement of atherosclerosis.

Test example 2 Effect of linseed oil on the intestinal flora of Atherosclerosis

1. Detection and analysis of intestinal flora

Fecal samples from control blank (CON), atherosclerosis Model (MOD) and linseed oil dry (MOD/FO) mice were collected separately using rnase-free EP tubes and sent to norstanding origin for 16S rRNA sequencing while the samples were frozen. Extracting the genomic DNA of the fecal sample by a hexadecyltrimethylammonium bromide (CT AB) method, detecting the purity and the concentration of DN A by agarose gel electrophoresis, taking a proper amount of sample in a centrifuge tube, and diluting the sample to 1 ng/. mu.L by using sterile water. Then, the diluted genomic DNA was used as a template, PCR-amplified with primers, enzymes and a buffer, and detected by electrophoresis using a 2% agarose gel. The samples were mixed in equal mass according to the concentration of the PCR product, and after mixing well, the PCR product was purified by electrophoresis using agarose gel with 1 XTAE concentration of 2%, and the target band was recovered by tapping (the product purification kit used was GeneJET gel recovery kit from Thermo Scientific). And finally, constructing a sequencing Library by using an Ion Plus Fragment Library Kit 48rxns Library construction Kit of the Thermofeisher company, and performing on-machine sequencing by using IonS5TMXL of the Thermofeisher company after the constructed sequencing Library is subjected to Qubit quantification and qualified Library detection.

As a result: firstly, the abundance and diversity of bacterial communities in each group of intestinal flora are analyzed by adopting alpha-diversity, and the intestinal flora is evaluated by observing species indexes and sparse curves. Referring to FIG. 2, A in FIG. 2 is the species index, and B is the dilution curve. Species index analysis showed that both the abundance and diversity of intestinal flora in the atherosclerosis model group (MOD) were altered compared to the placebo group (CON) (P < 0.05). By linseed oil dry prognosis, the abundance of intestinal flora of linseed oil dry pretreatment (MOD/FO) mice is changed, but the difference is not statistically significant (P < 0.05). The sparse curve can directly reflect the rationality of the sequencing data volume and indirectly reflect the abundance of the species in the sample, when the curve tends to be flat, the sequencing depth is basically covered to all the species in the sample, and as can be seen from fig. 2B, the curves of all groups of samples gradually tend to be flat along with the increase of the sequence number, the sequencing depth is basically covered to all the species in the sample.

To further verify the above results, unweighted UniFrac (PCoA) and weighted distance matrix (NMDS) and Wien diagrams were used to analyze the overall colony structure of the bacteria. Referring to fig. 3, in fig. 3, a is the PCoA result, B is the NMDS result, and C is the wien diagram. The PCoA analysis revealed a difference in intestinal flora between the atherosclerosis model group (MOD) and the placebo group (CON). The linseed oil pretreatment group (MOD/FO) also presented a different cohort compared to the atherosclerosis model group (MOD). At the same time, NMDS analysis showed the same results. The linseed oil intervention results in the change of the intestinal flora of the mice with atherosclerosis. The wien diagram shows that 269 kinds of bacteria are shared by a blank control group (CO N), an atherosclerosis model group (MOD) and a linseed oil dry stock group (MOD/FO), and 67, 70 and 25 kinds of bacteria are respectively specific to the blank control group (CON), the atherosclerosis model group (MOD) and the linseed oil dry stock group (MO D/FO).

The differential bacteria between the groups were further analyzed, and the differences in the flora structure of the mice of each group were compared at the phylum level. As shown in FIG. 4, A is the relative abundance of microorganisms at the phylum level, B is the ratio of firmicutes/bacteroidetes (F/B), and C is the phylum level difference bacteria (CON and MOD). It was found that at the phylum level, Firmicutes and Bacteroidetes constitute 2 common dominant phyla of all groups (fig. 4A). An increase in the firmicutes/bacteroidetes ratio (F/B) is strongly associated with obesity. The proportion of atherosclerosis model group (MOD) Firmicutes and Proteobacteria was significantly increased and the proportion of Bacteroidetes was significantly decreased (P <0.05) compared to the blank control group (CON). The ratio of firmicutes/bacteroidetes (F/B) of the atherosclerosis model group (MOD) is increased (P < 0.001). The linseed oil dry prognosis, with a markedly reduced firmicutes/bacteroidetes (F/B) ratio (P <0.001), indicates that linseed oil has a pronounced effect on the firmicutes/bacteroidetes (F/B) ratio in the intestinal tract of atherosclerotic mice under a high-fat diet.

On the genus level, the top 40 ranked species were analyzed. Referring to fig. 5, in fig. 5, a is the relative abundance of microorganisms at genus level, B is the comparison of genus level difference bacteria between the blank control group (CON) and the atherosclerosis model group (MOD), and C is the comparison of genus level difference bacteria between the atherosclerosis model group (MOD) and the linseed oil dry pre-group (MOD/FO). The results show that relative abundances of intestinomonas, Bilophila (cholephilus), anaerobotrucus (anoxyclavus), Oscillibacter, negavibacillus, blautia (blautita), parabia (paracoccus), muribacilum, paraeuterilla (paracasei), etc. were all increased (P <0.05), while relative abundances of alistes (cladia) and Candidat _ saccharas (candidate monosomycotiana) in the atherosclerosis model group (MOD) mice, compared to the blank control group (CON), indicate that the intestinal flora structure was altered in the atherosclerosis model group (MOD) mice. Relative abundances of Intestiminonas, Lachnoclostrium, Bilophila (Phalerochaete), Anaerotruncus (Anaerorhamus), Oscillbiter, Enterohabudus and Negatibacter in linseed oil dried group (MOD/FO) were all reduced compared to atherosclerosis model group (MOD) (P <0.05), indicating that linseed oil can down-regulate Intestiminonas, Lachnoclostrium, Bilophila (Phalerochaete), Anaerotruncus (Anaerorhamus), Oscilobacter, Enterohabudus and Negatibacter. The results show that the linseed oil can change the relative abundance of certain bacteria in the intestinal tracts of the atherosclerotic mice and has a regulating effect on the intestinal flora of the atherosclerotic mice.

2. Effect of linseed oil on metabolites of intestinal flora

The imbalance of intestinal flora can cause the level of flora metabolites to be changed, thereby influencing the pathogenesis process of atherosclerosis. Thus, after the linseed oil was found to be able to modulate the intestinal flora, the present assay further examined the effect of linseed oil on intestinal flora metabolites such as short chain fatty acids and bile acids.

2.1 Effect of Linseed oil on short-chain fatty acids as metabolites of intestinal flora

Short chain fatty acids were detected by Gas chromatography-mass spectrometer (GC-MS). TRACE 1310-ISQ LT gas chromatography-mass spectrometer of Thermo company in USA is adopted. The column was an Agilent HP-INNOWAX capillary column (30 m. times.0.25 mm ID. times.0.25 μm). Split-flow sample injection, the sample injection amount is 1 mu L, and the split-flow ratio is 10: 1. The injection inlet temperature is 250 ℃, the ion source temperature is 230 ℃, the transmission line temperature is 250 ℃, and the quadrupole rod temperature is 150 ℃. The temperature programming is started at 90 ℃, then the temperature is increased to 120 ℃ at 10/min ℃, then the temperature is increased to 150 ℃ at 5/min, and finally the temperature is increased to 250 ℃ at 25/min and maintained for 2 min. The carrier gas was helium and the flow rate of the carrier gas was 1.0 mL/min. Mass Spectrometry (MS) conditions: electron impact ionization (EI) source, SIM scan mode, electron energy 70 eV. Preparing a standard curve: weighing appropriate amount of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid and caproic acid standard substance, and preparing into eleven standard concentration gradients of 0.01, 0.1, 0.5, 1, 5, 10, 25, 50, 100, 250 and 500 mug/mL respectively by using diethyl ether. And (3) respectively carrying out GC-MS detection on the concentration series of the standard solution, carrying out linear regression analysis by taking the concentration (X) of the standard substance as an abscissa and taking the peak area ratio (Y) of the standard substance to the internal standard substance as an ordinate, and drawing a standard curve. And simultaneously, the precision, the repeatability, the recovery rate and the quantitative limit are investigated.

As a result: referring to fig. 6, a is a short chain fatty acid peak plot and B is a differential short chain fatty acid clustering heat map in fig. 6. The short chain fatty acids of each single peak were clearly resolved from the short chain fatty acid peak plot, indicating that the method and data are reliable. Clustering heat maps indicated differences in short chain fatty acids in the placebo (CON), atherosclerosis Model (MOD) and linseed oil pretreatment (MOD/FO) groups. Further, the effect of linseed oil on differential short chain fatty acid content was analyzed. As shown in fig. 7, a in fig. 7 is the acetic acid content result, B is the propionic acid content result, C isovaleric acid content result, D isobutyric acid content result, E valeric acid content result. As can be seen in fig. 7, the levels of acetic acid, propionic acid and valeric acid were decreased in the atherosclerosis model group (MOD) compared to the blank control group (CON) (P < 0.05). Compared with the atherosclerosis model group (MOD), the linseed oil dry stock group (MOD/FO) has increased contents of acetic acid, propionic acid, isovaleric acid, isobutyric acid and valeric acid (P <0.05), and other short-chain fatty acids have no significant change. The above results indicate that dysbacteriosis of the intestinal tract caused by atherosclerosis leads to the reversal of the abnormally reduced metabolite short-chain fatty acids under the intervention of linseed oil.

2.2 Effect of Linseed oil on intestinal flora metabolite Bile Acids (BAs)

Bile acids, another metabolite of intestinal microorganisms, have also been reported to be associated with the progression of chronic metabolic disease. Therefore, the effect of linseed oil on bile acids was determined by liquid chromatography-mass spectrometer (LC-MS). First, 38 standard solutions were prepared for detecting the composition of bile acids, and the prepared standard solutions were stored at-20 ℃ for later use. Collecting feces samples of mice of blank control group (CON), atherosclerosis model group (MOD) and linseed oil dry pre-treatment group (MOD/FO) about 100mg each, adding 0.3mL methanol precipitate protein, vortex shaking for 1min, and centrifuging at 4 deg.C for 10min (12000 Xg); the supernatant was diluted 10-fold, vortexed and shaken for 1min, centrifuged at 4 ℃ for 10min (12000 Xg), and the supernatant was aspirated for LC-MS analysis. By using ACQUITYBEH C18 chromatographic column (2.1X 100mm, 1.7 μm, Waters corporation, USA) with sample amount of 5 μ L, column temperature of 40 deg.C, mobile phase A-0.01% formic acid water, B-acetonitrile. Gradient elution conditions are 0-4 min, and 25% of B; 25-30% B for 4-9 min; 9-14 min, 30-36% of B; 14-18 min, 36-38% B; for 18-24 min, 38-50% of B; 24-32 min, 50-75% B; 32-35 min, 75-100% B; 35-38 min, 100-25% B. The flow rate was 0.25 mL/min. Mass spectrum conditions: electrospray ionization (ESI) source, negative ion ionization mode. The ion source temperature is 500 ℃, the ion source voltage is-4500V, the collision gas is 6psi, the gas curtain gas is 30psi, the atomization gas and the auxiliary gas are both 50psi, and finally, Multiple Reaction Monitoring (MRM) is adopted for scanning.

As a result: referring to fig. 8, in fig. 8, a is a bile acid peak graph, and B is a differential bile acid clustering heat map. The bile acid of each single peak can be clearly resolved from the bile acid peak diagram, indicating that the method and data are reliable. The cluster heatmap shows the differences in bile acid content between the atherosclerotic disease model group (MOD) and the linseed oil pretreatment group (MOD/FO). As can be seen from fig. 9, after linseed oil drying, the contents of alloLCA (allolithocholic acid), isoLCA (isocholic acid), 7-ketoLCA (7-ketolithocholic acid), β -UDCA (3 β -ursodeoxycholic acid), CDCA (chenodeoxycholic acid) and HDCA (hyodeoxycholic acid) were all decreased (P <0.05), while the contents of LCA (lithocholic acid), ACA (allocholic acid), GCA (glycocholic acid) and TCA (taurocholic acid) were all increased. Indicating that linseed oil can change the bile acid level of the metabolite of the intestinal flora.

As can be seen from the above test examples, the atherosclerosis mouse model of the invention is prepared by linseed oil drying, the method comprises the steps of detecting aorta and aortic sinus lipid plaque conditions and arterial vessel fibrosis level through histological staining, analyzing abundance and diversity change of mouse intestinal flora through 16S rRNA detection, respectively detecting changes of short-chain fatty acid and bile acid of metabolites of the intestinal flora through gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry, finding that linseed oil can obviously slow down lipid deposition of atherosclerotic vascular plaque, and can down-regulate Intestimonas, Bilophila (cholephilus), Anaerotrycus (anaerobic clavatus), Oscillbacter, Negatibacter, Lachnoclostridium and Enterorhabdus in the intestinal flora of the mouse with atherosclerosis, and the levels of short-chain fatty acid as a metabolite and bile acid as a metabolite of intestinal flora are changed, which indicates that the linseed oil can improve atherosclerosis by regulating the intestinal flora. Therefore, the linseed oil can be used for preparing a medicament for improving the intestinal flora of atherosclerosis so as to relieve the development of atherosclerosis. The medicine comprises effective dose of linseed oil and pharmaceutically acceptable auxiliary materials or auxiliary components. The medicine can be prepared into tablets, capsules, lotion, granules or oral preparations according to a conventional preparation method.

While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.