CN116870101A - Application of monascus fermentation product in regulating lipid metabolism - Google Patents

Abstract

The invention relates to the technical field of biological medicine, and provides application of a monascus fermentation product in regulating lipid metabolism.

Description

Application of monascus fermentation product in regulating lipid metabolism

Technical Field

The invention relates to the technical field of biological medicines, in particular to application of a monascus fermentation product in regulating lipid metabolism.

Background

With the economic development and the improvement of the living standard of people, the obesity and related metabolic diseases caused by high-fat diet, sedentary life style and other life modes are increasing, and the hyperlipidemia is one of them. Hyperlipidemia can cause fat metabolism disorder and abnormal blood lipid transport, so that cholesterol, triglyceride and low density lipoprotein in blood plasma are increased, high density lipoprotein is reduced, hyperlipidemia is closely related to oxidative stress, and cell damage is easily caused. Hyperlipidemia is a metabolic disorder disease, which causes disturbance of fat metabolism and abnormal blood lipid transport and is closely related to oxidative stress. The monascus can utilize the substrate to generate a large amount of biological functional metabolites including monascus pigment, monacolin K and the like, and the secondary metabolites have the functions of improving hyperlipidemia, hypertension, oxidative stress and the like. At present, more and more researches prove that the monascus fermentation product can reduce diet-induced hypercholesteremia, and lipid-lowering metabolites of the monascus play an important role in the lipid-lowering market. Monacolin K is the main active ingredient of the monascus fermentation product, and exists in two forms of acid type (ring opening) and lactone type (ring closing), wherein the stability of the lactone type Monacolin K is higher than that of the acid type Monacolin K. Under the acidic condition, the acid type monacolin K is continuously converted into lactone type, and the lactone type monacolin K has the activity of reducing blood fat and cholesterol. 5-7 mg of monacolin K has the same lipid-lowering effect as 20-40 mg of pure lovastatin.

Currently, cereal matrixes are mainly used as fermentation matrixes of monascus, and the cereal matrixes have higher economic value, so that the cost is higher, and the mass production is not facilitated. The production rate of Monascus for producing Monacolin K is not high by fermenting on cereal matrix, which is unfavorable for large-scale production and mass production of enterprises.

The existing medicines for improving the hyperlipidemia mainly comprise the following two main types: firstly, statin drugs mainly comprise simvastatin, lovastatin, atorvastatin and the like, wherein an open acid part in the statin drugs is similar to the structure of hydroxymethyl glutarate coenzyme A which is a speed limiting enzyme for synthesizing cholesterol by the liver, the statin drugs have competitive inhibition effect on the enzyme, and the cholesterol concentration can be effectively reduced after the statin drugs are taken; secondly, fibrate drugs mainly comprise ciprofibrate, bisfibrate and benzalkonium Bei Tedeng, and the purposes of reducing lipid are achieved by promoting the liver to absorb fatty acid and inhibiting the liver from synthesizing triglyceride, activating a peroxisome proliferator-activated receptor, inducing lipoprotein esterase expression and promoting triglyceride hydrolysis in lipoprotein particles.

Statin and fibrate drugs are generally expensive and can be associated with adverse reactions in the gastrointestinal tract, liver hypofunction, etc. for long-term administration. The statin drugs are needed to participate in hydrolysis by hydroxy acid esterase in human body when the statin drugs act, and after the statin drugs are ingested by the human body, liver injury and myopathy can be caused, and digestive tract symptoms such as headache, insomnia, depression, dyspepsia, diarrhea, abdominal pain, nausea and the like can also be caused.

Disclosure of Invention

The invention aims to provide a novel application of monascus fermentation products in regulating lipid metabolism. The invention mainly designs the dynamic change of the monacolin K produced by fermenting the monascus and the animal experiment of the monacolin K, and explores the influence of the monacolin K on lipid metabolism of a high-fat diet mice by taking the pomelo ascus as a fermentation matrix.

In order to solve the technical problems, the invention adopts the following technical scheme:

the application of monascus fermentation product in pharmacy is used for regulating lipid metabolism.

The invention selects monascus which can metabolize and produce Mongolin K for fermentation, and aims at relieving diet-induced hypercholesteremia by using the natural fermentation product Mongolin K of the monascus. The red yeast rice is used in human diets since ancient times, and natural compounds extracted from common edible fungi can avoid side effects caused by chemically synthesized statin drugs in human bodies and avoid damage to human livers and kidneys.

Experiments show that the monascus fermentation product can be used for regulating lipid metabolism of high-fat diet mice, reducing the blood lipid concentration of the high-fat mice, influencing the expression of genes related to liver and lipid metabolism of the mice, and the technical result can provide basis for further development and utilization of monascus fermentation shaddock bag products.

The fungus is cultured to produce bioactive substances for disease prevention and treatment, the low-cost shaddock byproduct, namely shaddock bag, is used for fermenting monascus, and the fermented product is further used for regulating lipid metabolism of high-fat diet mice and reducing the blood lipid concentration of the high-fat mice.

Preferably for reducing fat accumulation in organs.

Preferably, the organ comprises one or more of liver and epididymis.

Preferably, for alleviating liver metabolic disorders.

Preferably, it is used to inhibit the deposition of one or more of cholesterol, triglycerides, bile acids in the liver.

Preferably, it is used for reducing triglyceride content in serum.

Preferably for reducing accumulation of low density cholesterol in blood.

Preferably, for inhibiting cholesterol synthesis.

Preferably, it is used for accelerating the discharge of cholesterol synthesis bile acid from the body.

A method for preparing the monascus fermentation product comprises the following steps:

A. inoculating monascus into a seed culture medium to obtain seed liquid; the seed medium comprises: glucose 60g/L, fish meal peptone 25g/L, naNO ₃ 2g/L, dipotassium hydrogen phosphate 1g/L, mgSO ₄ ·7H ₂ O1g/L, corn starch 6g/L, and adjusting the pH to 6.

B. Inoculating the seed solution into a shaddock bag with an inoculum size of 10% (mL/g), and adjusting the initial water content to 45%; fermenting for more than 16 days at the temperature of 25-30 ℃ to obtain the monascus fermentation product.

The pomelo ascus is a common processing byproduct of the fruit pomelo in the south, is rich in pectin, vitamins, saccharides and other substances, but most of the pomelo ascus is discarded, and only a small part of the pomelo ascus is further processed into leisure snack products, so that the pomelo ascus has fewer audiences. The invention uses the common and low-price shaddock processing byproduct in the south, namely the shaddock bag, to replace cereal substances as the fermentation matrix of monascus, and explores the influence of the shaddock bag after the monascus fermentation on lipid metabolism of the hyperlipoidemia mice.

The invention selects the low-cost shaddock byproduct, namely the shaddock bag, as the fermentation substrate of the monascus, explores the lipid metabolism function effect of oral monascus fermented and unfermented shaddock bag samples on high-fat diet mice while reducing the fermentation cost of the monascus, and provides a usable reference for further development and utilization of monascus fermented shaddock bag products.

The invention selects the pomelo ascus as the fermentation substrate of the monascus, and remarkably improves the yield of the monascus metabolite monacolin K.

The invention selects the byproduct shaddock asc of shaddock as the fermentation substrate, improves the utilization rate of shaddock capsules, reduces waste after fruit processing, replaces cereal substrates commonly used in the market at present with low-price shaddock capsules, and effectively reduces the fermentation cost of monascus.

Compared with the prior art, the implementation of the invention has the following beneficial effects:

1. the monascus fermentation metabolite contains the component capable of reducing the blood lipid concentration, namely monacolin K, and when the monacolin K obtained by a natural microorganism fermentation way reduces the blood lipid concentration, hydroxy acid esterase in a human body is not needed to participate in hydrolysis, so that side effects such as adverse reactions of gastrointestinal tract, liver and kidney hypofunction and the like caused by taking lipid-lowering chemical synthetic drugs can be avoided.

2. The common cereal matrixes are selected for fermenting monascus in the market at present, so that the production cost is high, and the shaddock processing byproduct shaddock ascus which is low in price and common in the south is selected as the solid fermentation matrix of the monascus, so that the utilization rate of agricultural products is improved, and meanwhile, the fermentation cost of the monascus is reduced.

3. The nutritional ingredients of the naringin can effectively improve the yield of the monascus metabolites and the yield of the monacolin K.

4. The monascus pomelo bag fermented product can be used for regulating lipid metabolism of high-fat diet mice, reducing the blood lipid concentration of the high-fat mice, and influencing the expression of genes related to lipid metabolism in the livers of the mice.

Drawings

FIG. 1 is a schematic diagram showing the change of substances during fermentation: variation of shaddock bag dry weight in monascus solid state fermentation process. The change of Monascus K in the solid state fermentation process. Results are expressed as mean ± SD (n=3).

Fig. 2 is a schematic diagram showing the effect of monascus fermented pomelo bag powder on physiological index and epididymal fat accumulation parameters of mice in each group: (a) a body weight growth curve, (B) a liver index, (C) an epididymal fat index, and the results are expressed as mean ± SD (n=8). Statistical analysis was performed using one-way analysis of variance (ANOVA). Significant differences from model set: * p <0.05, < p <0.01, < p <0.001, < p <0.0001.

FIG. 3 is a schematic of lipid levels in serum: the composition comprises (A) cholesterol content, (B) triglyceride content, (C) low-density lipoprotein cholesterol content, (D) high-density lipoprotein cholesterol content, (E) glutamic-oxaloacetic transaminase content, and (F) glutamic-pyruvic transaminase content. Results are expressed as mean ± SD (n=8). Statistical analysis was performed using one-way analysis of variance (ANOVA). Significant differences from model set: * p <0.05, < p <0.01, < p <0.001, < p <0.0001, ns have no significant difference.

Fig. 4 is a schematic representation of lipid levels in the liver: the content of (A) cholesterol, (B) triglyceride, and (C) bile acid. Results are expressed as mean ± SD (n=8). Statistical analysis was performed using one-way analysis of variance (ANOVA). Significant differences from model set: * p <0.05, < p <0.01, < p <0.001, < p <0.0001, ns have no significant difference.

Fig. 5 is a schematic representation of the change of oxygen radicals in the liver: the glutathione content, (B) the catalase activity, (C) the superoxide dismutase activity, and (D) the malondialdehyde content. Results are expressed as mean ± SD (n=8). Statistical analysis was performed using one-way analysis of variance (ANOVA). Significant differences from model set: * p <0.05, < p <0.01, < p <0.001, < p <0.0001, ns have no significant difference.

Fig. 6 is a schematic representation of the liver histopathological observations of mice: (A) control group, (B) model group, (C) UF-L group, (D) UF-H group, (E) F-L group, and (F) F-H group.

FIG. 7 is a schematic diagram of metabonomic analysis of mouse liver: (A) PCA score plots of liver samples integrated with metabonomics data. (B) PLS-DA score plots of liver samples integrated with metabonomics data. OPLS-DA score plots for model group (C) and F-H group. (D) Volcanic plot of metabonomics data for model and F-H groups.

FIG. 8 is a schematic representation of expression of cholesterol metabolism related genes in liver tissue: (A) CYP7A1 gene expression level, (B) LDLr gene expression level, (C) HMGCR gene expression level, and (D) SREBP-1C gene expression level. Results are expressed as mean ± SD (n=8). Statistical analysis was performed using one-way analysis of variance (ANOVA). Significant differences from model set: * p <0.05, < p <0.01, < p <0.001, < p <0.0001, ns have no significant difference.

Detailed Description

The present invention will be further described in detail below with reference to the accompanying drawings, for the purpose of making the objects, technical solutions and advantages of the present invention more apparent, so that those skilled in the art can better understand the present invention and implement the present invention, but the present invention is not limited thereto. Modifications and substitutions to methods, procedures, or conditions of the present invention without departing from the spirit and nature of the invention are intended to be within the scope of the present invention. The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated.

Example 1

1. Monascus solid state fermentation pomelo ascus producing monacolin K

The experimental steps are as follows:

fermentation conditions of monascus solid state fermentation of the pomelo ascopecurs:

inoculating Monascus to a seed culture medium to obtain seed solution, and culturing at 30deg.C and 180rpm for 2 days to obtain seed solution; the seed medium comprises: glucose 60g/L, fish meal peptone 25g/L, naNO ₃ 2g/L, dipotassium hydrogen phosphate 1g/L, mgSO ₄ ·7H ₂ O1g/L, corn starch 6g/L, and adjusting the pH to 6.

The solid fermentation medium consisted of 10g of dried pomelo sacs. The initial water content of the culture medium is adjusted to 45% before fermentation, the pH value is natural, the culture medium is stirred uniformly, sterilization is carried out for 30min at 121 ℃, and then seed liquid is inoculated. The culture conditions are that fermentation is carried out for 2d at 30 ℃, and the fermentation is carried out for 12, 14, 16, 18, 20, 22 and 24d at 25 ℃, so that samples are prepared, and the content of the monacolin K in the fermented shaddock bag product is detected. The fermented and unfermented dried shaddock bag samples are respectively crushed into powder by a crusher, diluted by water and used for mice. 5g of each weighed powder is placed in a 50mL cuvette and the volume is fixed to 50mL by 70% ethanol solution. The reaction was carried out in a thermostatic water bath at 55℃for 1h, shaking every 20 min. Cooling to room temperature, and filtering with 0.22 μm organic filter membrane to obtain 100 g.L ^-1 And (3) extracting the liquid from the sample.

High performance liquid chromatography analysis of monacolin K: the Monacolin K assay uses a Waters2487 series model 1525 HPLC system (Waters, 20RBA x SB-C18 column) and a UV detector. And calculating the contents of the two substances according to the lactone type Monacolin K and the acid type Monacolin K standard curve, and adding the contents to obtain the total content of the Monacolin K. The experimental results are shown in FIG. 1.

As can be seen from fig. 1: as shown in fig. 1, the dry weight slightly increased on the first day, probably due to the increase in dry weight caused by hyphae growth. There was no significant change in dry weight between day 2 and day 4, probably due to the relative balance of the growth rate of hyphae and the substrate consumption rate of the naringena. Between days 4-16, the dry weight of the pomelo sacs was significantly reduced, indicating that they were consumed in large amounts, at rates exceeding the hyphal growth rate. It is also possible that the pomelo capsule is mainly used for synthesizing secondary metabolites rather than promoting hypha growth. After day 16, the dry weight remained essentially unchanged, indicating that the fermentation process was complete. Overall, the dry weight of the post-fermentation pomelo sacs was reduced by 32.1% compared to before fermentation. The variation of the K content of monacolin throughout the fermentation is shown in FIG. 1. On the first day of fermentation, the content of monacolin K was zero, indicating that the naringin ascmatrix itself did not have monacolin K, and that a small amount of monacolin K was produced between days 2 and 4. Over time, the K content of monacolin tended to rise, reaching a peak value after 16 days, and stabilizing at 6816.2 mg.kg ^-1 . In the whole fermentation process, the change of the content of the monacolin K is identical with the change of the dry weight of the shaddock bag in the culture medium, which shows that the monascus can utilize the shaddock bag to carry out growth metabolism and the production of the monacolin K.

Example 2

Design of animal experiment

72 male mice (20.+ -.2 g) of 6 weeks old were purchased from Wu's laboratory animal trade company (China, fujian). Rats were randomly divided into 6 groups after 7d of adaptive feeding: control group (standard normal feed + distilled water), model group (high fat feed + distilled water), UF-L group (high fat feed + low dose pomelo capsule product), UF-H group (high fat feed + high dose pomelo capsule product), F-L group (high fat feed + low dose red yeast fermented pomelo capsule product) and F-H group (high fat feed + high dose red yeast fermented pomelo capsule product). Mice were perfused with 0.2mL daily and continuously for 8 weeks, and body weights were measured at the same time weekly. 1d before the end of the experiment, the experimental animals fasted for 12h. Blood is collected by adopting an eyeball blood taking mode, and important tissues of liver and epididymis are rapidly taken. All procedures were in compliance with current guidelines for laboratory animal care and use.

Standard normal feed, available from wu's laboratory animal trade limited. High fat feeds were purchased from south Tong Telofei feed technologies Inc., and used with reference to the feed feeding methods provided by the company.

Example 3

Mouse organ index detection

The experimental steps are as follows: the liver and epididymal fat of the mice were washed with phosphate buffer PBS and the surface water was blotted off with filter paper. Body weight was measured and organ index = organ weight/mouse body weight 100%.

The experimental results are shown in FIG. 2.

As can be seen from fig. 2: at the beginning of the experiment, the initial body weights of the mice in each group were not different. In the experimental process, the body weight of each group of experimental mice is in a growing trend, and the body weight of the model group is obviously higher than that of other groups. The weights of the mice in the UF-L group, the UF-H group and the F-L group are lower than that of the model group and higher than that of the control group, and the weights of the mice in the F-H group are not obviously different from that of the control group, so that the monascus fermented pomelo bag powder has obvious inhibiting effect on weight increase and obesity of the mice. The liver index and epididymal fat index of each group of mice are shown in fig. 2B and 2C. The liver indexes and epididymal fat indexes of the UF-L group, the UF-H group, the F-L group and the F-H group are lower than those of the model group, and the reduction of the liver indexes and epididymal fat indexes of the F-H group and the epididymal fat indexes of the F-H group is more obvious compared with those of other groups, so that the accumulation of liver and epididymal fat can be effectively reduced by using the monascus fermented shaddock bag powder, and the effect of the high-dose monascus fermented shaddock bag powder is better.

Example 4

Serum biochemical index detection

The experimental step comprises centrifuging blood sample at 4000r/min for 10min, collecting supernatant, and detecting serum biochemical index, and detecting Total Cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) by using full-automatic biochemical analyzer. The activity of glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) was detected with a kit (Nanjing institute of biotechnology, china).

The experimental results are shown in FIG. 3.

As can be seen from fig. 3: compared with the control group, the serum Total Cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) levels of the mice in the model group are obviously increased, and the content of high-density lipoprotein cholesterol (HDL-C) is obviously reduced. Compared with the model group, the levels of total cholesterol, triglyceride, low density lipoprotein cholesterol, glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase in the serum of the mice of the UF-L group, the UF-H group, the F-L group and the F-H group are reduced, and the content of high density lipoprotein cholesterol is increased. Wherein the effects of the UF-H group, the F-L group and the F-H group are remarkable, and the effects of the F-H group are more remarkable. The triglyceride content and glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase levels of the serum of the F-H group mice are even lower than those of the control group, which indicates that the high-dose monascus fermented pomelo bag powder has better effect of improving blood lipid metabolism, has a protective effect on liver and can effectively improve liver injury.

Example 5

Liver biochemical index detection

The experimental steps are as follows: taking liver tissue 100mg, placing into a 2mL centrifuge tube, washing with physiological saline to remove surface impurities, and adding physiological saline according to the weight ratio of 1:9. Liver tissue was minced with scissors and then inserted into ice. Further disrupting the tissue with a tissue homogenizer, centrifuging at 9500r/min for 5min, and collecting supernatant for biochemical index detection including Total Cholesterol (TC), triglyceride (TG), catalase (CAT), and Glutathione (GSH). Malondialdehyde (MDA), superoxide dismutase (SOD) and Bile Acids (BAs) were measured with a kit (Nanjing institute of biological engineering, nanjing, jiangsu, china).

The experimental results are shown in fig. 4 and 5.

As can be seen from fig. 4: the cholesterol, triglyceride and bile acid contents in liver are shown in figure 4. Compared with the control group, the liver of the mice in the model group has obviously increased cholesterol, triglyceride and bile acid content. Compared with the model group, the cholesterol, triglyceride and bile acid contents in the livers of the mice in the UF-L group, the UF-H group, the F-L group and the F-H group are reduced, the triglyceride, the cholesterol and the bile acid contents in the livers of the mice in the F-L group and the F-H group are obviously reduced, and the reduction effect of the F-H group is more obvious, which indicates that the high-dose red yeast fermented pomelo bag powder can more effectively remove TG, TC and BAs in the livers and reduce the deposition of cholesterol, triglyceride and bile acid in the livers.

The content of malondialdehyde can reflect the degree of lipid peroxidation and thus indirectly the degree of hepatocyte damage. Superoxide dismutase activity represents the ability of an organism to detoxify oxygen radicals. Glutathione (GSH) is an active ingredient of reduced glutathione, and participates in the oxidation-reduction process in vivo, can be combined with peroxide and free radicals, and can resist the damage of the free radicals to important organs. Catalase (CAT) can promote H ₂ O ₂ To break down into molecular oxygen and water, scavenge hydrogen peroxide in the body, and protect cells from H ₂ O ₂ Is a toxic hazard. As can be seen from fig. 5, the superoxide dismutase and catalase activities in the liver of the mice of the model group were reduced and the glutathione and malondialdehyde contents were increased as compared with the control group. Comparing with the model group, UF-L group,The activities of superoxide dismutase and catalase in the livers of the mice in the UF-H group, the F-L group and the F-H group are increased, and the contents of glutathione and malondialdehyde are reduced. The F-H group has remarkable effect on improving the activities of superoxide dismutase and catalase and reducing the contents of glutathione and malondialdehyde, and is close to the level of a control group, so that the high-dose monascus fermented pomelo bag powder can effectively improve the activities of superoxide dismutase and glutathione in the liver, simultaneously clear oxygen free radicals, reduce the content of malondialdehyde, inhibit lipid peroxidation and improve the antioxidant capacity, thereby reducing the oxidative damage of the liver.

Example 6

Liver histopathological evaluation

The experimental steps are as follows: after the liver tissues of each group of mice were fixed with 10% formaldehyde solution, paraffin-embedded, sectioned, hematoxylin and eosin (H & E) stained and observed under an optical microscope.

Experimental results: each set of liver tissue slices is shown in fig. 6.

As can be seen from fig. 6: the liver cells of the control group are orderly, regular and compact in arrangement and normal in morphology, and no obvious fat drops are found. The liver cells of the model group are abnormal in state, disordered in arrangement, larger in cell gap, balloon-like in liver cell volume, serious in liver injury and large in fat drop in cells. The overall morphology of the hepatocytes of UF-L, UF-H, F-L and F-H groups was improved to a different extent than that of the model group. The improvement effect of the liver of the F-H group is more obvious, the quantity of fat drops is obviously reduced, the cell arrangement is relatively tidy, the liver tissue state is optimal, and the effect is close to that of a control group.

Example 7

Mass spectrometry of liver samples

The experimental steps are as follows: liver samples (25 mg) were mixed with 1mL of extract (acetonitrile: methanol: water=2:2:1), homogenized, sonicated, and centrifuged at 12000×g for 10min at 4 ℃. The supernatant was dried in a vacuum concentrator at 37 ℃. The dried sample was reconstituted with 200 μl of 50% acetonitrile. Centrifuge at 14000rpm for 15min at 4℃and filter the supernatant through a 0.22 μm filter for UPLC-MS/MS analysis. UPLC separation A1290 Infinicity series UPLC System (Agilent Technologies) was used, equipped with a UPLC BEH Amide column (2.1X100 mm,1.7 μm, waters). The mobile phase was 25mmol/L of an aqueous solution of ammonium acetate and aqueous ammonia (A) and acetonitrile (B). The sample determination gradient elution procedure was: 0 to 0.5min,95 percent of B;0.5 to 7.0min,95 to 65 percent of B;7.0 to 8.0min,65 to 40 percent of B;8.0 to 9.0min,40 percent of B;9.0 to 9.1min,40 to 95 percent of B; the column temperature was 25 ℃, the autosampler temperature was 4 ℃, and the sample injection amount was 2. Mu.L.

Raw UPLC-QTOF/MS data was processed using Mass Lynx 4.1 (Waters, USA). Principal Component Analysis (PCA), partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were performed with "factor extra" using R3.5.1 software.

The experimental results are shown in FIG. 7.

As can be seen from fig. 7: the liver plays the most important role in substance metabolism and energy metabolism and is a source of a large number of endogenous metabolites. The sample metabolite distribution among the groups is shown in figure 7. In the PCA scoring chart 7A, the points in different groups are relatively gathered, which means that the individual differences in the groups are smaller, and the repeatability in the groups is better, and the sample data are very similar. Specific differences in separation between groups require further PLS-DA analysis. The data sets of Control, model, UF-H and F-H groups were analyzed using PLS-DA, and in FIG. 7B, a clear separation between Control and other groups was observed, with the data points of UF-H and F-H groups gathered between the Control and Model groups, the F-H and Control groups having the same trend, and the F-H and Model groups being significantly separated. In FIG. 7C, there was a clear difference between the hepatic metabolites of the F-H group and the Model group, indicating that the Model group interfered with hepatic metabolism. Volcanic figure 7D further demonstrates this, and the results indicate that the F-H group has 182 total differential metabolites compared to the Model group, with 85 up-regulated and 97 down-regulated metabolites, indicating complex systemic changes in mouse liver metabolites. The result shows that the liver metabolite of the MRPS dry prognosis high-fat diet mice deviates from the model group, is closer to the control group, gradually approaches to the liver metabolite of the normal mice, and obviously relieves the liver metabolic disorder of the mice.

Example 8

RT-PCR analysis

The experimental steps are as follows: and detecting the relative expression quantity of mRNA by adopting real-time fluorescence quantitative RT-PCR. UsingTotal liver RNA was extracted using the TotalRNAkit (omega Bio-TekInc, GA, USA). RNA samples were used to evaluate a range of genes thought to be involved in cholesterol metabolism, including low density lipoprotein receptor (LDLr), cholesterol regulatory element sterol binding protein (SREBP-1 c), 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR), and cholesterol 7-alpha-hydroxylase (CYP 7A 1). The primer sequences are shown in Table 1. Relative quantification of expression of each gene in liver was 2 ^-ΔΔCt 。

TABLE 1 primers for RT-PCR

The experimental results are shown in FIG. 8.

As can be seen from fig. 8: compared with a control group, the expression quantity of CYP7A1 and LDLr in the liver is obviously reduced, the conversion of cholesterol is inhibited, the expression quantity of HMGCR and SREBP-1c in the model group is obviously improved, the generation of cholesterol is increased, and the accumulation of cholesterol in blood is caused. In the UF-L group, the UF-H group, the F-L group and the F-H group, the expression quantity of CYP7A1 and LDLr is obviously improved, the expression quantity of HMGCR and SREBP-1c is obviously reduced, wherein the effect of the F-H group is obvious, and the high-dose monascus fermented pomelo bag powder is proved to be capable of effectively reducing accumulation of low-density cholesterol in blood, inhibiting synthesis of cholesterol, increasing discharge of cholesterol synthetic bile acid and greatly reducing concentration of cholesterol in a body.

Example 9

From the above experimental results, it can be seen that:

1. according to the invention, the pomelo ascus is used as the monascus fermentation matrix to replace the common cereal matrix in the market, so that the utilization rate of the pomelo ascus is increased, and the waste is reduced. Meanwhile, the naringin is adopted as the monascus fermentation substrate, so that the yield of the Monacolin K is greatly increased to 6816.2 mg.kg ^-1 。

2. According to the invention, the model and the control mice are fed with the monascus fermented and unfermented dried pomelo bag powder respectively, and the effect are observed, and the organ indexes of the mice are measured, so that the feeding of the monascus fermented pomelo bag powder has obvious inhibiting effect on weight increase and obesity of the mice, can effectively reduce accumulation of liver and epididymis fat, has better effect of improving blood lipid metabolism, has protecting effect on the liver, and can effectively improve liver injury. The existing lipid-lowering drugs in the market have a certain damage effect on the liver, and the shaddock bag fermented by monascus in the invention has the effect of improving the liver damage.

3. The invention proves that the high-dose monascus fermented pomelo bag powder can more effectively inhibit the deposition of triglyceride, cholesterol and bile acid in the liver by exploring the effect of the monascus fermented and unfermented dried pomelo bag powder on the liver of a mouse; the liver antioxidant capacity analysis experiment proves that the high-dose monascus fermented pomelo ascus powder can effectively improve the activities of superoxide dismutase and glutathione in the liver, simultaneously clear oxygen free radicals, reduce malondialdehyde content, and inhibit lipid peroxidation, thereby reducing oxidative damage of the liver; pathological analysis of liver tissue can observe that the improvement effect of the F-H liver is more obvious, the quantity of fat drops is obviously reduced, the cell arrangement is relatively orderly, the liver tissue state is optimal, and the effect is close to that of a control group.

4. The invention also explores the influence of monascus fermented and unfermented dried shaddock bag sample powder on the expression of genes related to lipid-lowering function in liver. The expression level of CYP7A1 and LDLr in the liver is obviously reduced by the model group, the conversion of cholesterol is inhibited, the expression level of HMGCR and SREBP-1c in the model group is obviously improved, the generation of cholesterol is increased, and the accumulation of cholesterol in blood is caused. In the UF-L group, the UF-H group, the F-L group and the F-H group, the expression quantity of CYP7A1 and LDLr is obviously improved, the expression quantity of HMGCR and SREBP-1c is obviously reduced, the conversion rate of cholesterol is improved, and the accumulation quantity is reduced, wherein the effect of the F-H group is obvious.

The foregoing disclosure is merely illustrative of the preferred embodiments of the present invention and is not intended to limit the scope of the claims herein, as equivalent changes may be made in the claims herein without departing from the scope of the invention.

Claims (10)

1. The use of a monascus fermentation product in the manufacture of a medicament for modulating lipid metabolism.

2. The use according to claim 1 for reducing fat accumulation in organs.

3. The use of claim 2, wherein the organ comprises one or more of a liver, epididymis.

4. The use according to claim 1 for alleviating a metabolic disorder of the liver.

5. The use according to claim 1 for inhibiting the deposition of one or more of cholesterol, triglycerides, bile acids in the liver.

6. Use according to claim 1 for reducing the triglyceride content in serum.

7. The use according to claim 1 for reducing accumulation of low density cholesterol in blood.

8. The use according to claim 7 for inhibiting cholesterol synthesis.

9. The use according to claim 7 for accelerating the removal of cholesterol synthesis bile acids from the body.

10. A process for the preparation of a monascus fermentation product as claimed in claim 1, comprising the steps of:

A. inoculating Monascus to seed culture medium, and culturing at 30deg.C and 180rpm for 2 days to obtain seed solution; the seed medium comprises: glucose 60g/L, fish meal peptone 25g/L, naNO ₃ 2g/L, dipotassium hydrogen phosphate 1g/L, mgSO ₄ ·7H ₂ O1g/L, corn starch 6g/L, and regulating the pH to 6;

B. inoculating the seed solution into a shaddock bag with an inoculum size of 10% (mL/g), and adjusting the initial water content to 45%; fermenting for more than 16 days at the temperature of 25-30 ℃ to obtain the monascus fermentation product.