CN114028524B - Compound for improving blood lipid metabolism and preparation method and application thereof - Google Patents

Compound for improving blood lipid metabolism and preparation method and application thereof Download PDF

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CN114028524B
CN114028524B CN202111480443.1A CN202111480443A CN114028524B CN 114028524 B CN114028524 B CN 114028524B CN 202111480443 A CN202111480443 A CN 202111480443A CN 114028524 B CN114028524 B CN 114028524B
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lipid metabolism
blood lipid
lactobacillus plantarum
improving blood
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CN114028524A (en
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文兴建
刘晏齐
邢雅婧
钟亚东
胡娟
刘莉
刘周江
陈强
罗晓玲
侯凌云
黎敏
陈建康
王明玉
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Chongqing Academy of Chinese Materia Medica
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    • A61K36/704—Polygonum, e.g. knotweed
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    • A61K36/894—Dioscoreaceae (Yam family)
    • A61K36/8945—Dioscorea, e.g. yam, Chinese yam or water yam
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment

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Abstract

The invention relates to the field of health-care product methods, and particularly discloses a compound for improving blood lipid metabolism and a preparation method and application thereof, wherein the compound is prepared from the following components in parts by weight: 20-30 parts of radix curcumae, 10-12 parts of turmeric, 6-8 parts of Chinese yam, 4-6 parts of hawthorn, 4-6 parts of giant knotweed and 1-2 parts of microbial additive, wherein the microbial additive is prepared from lactobacillus plantarum subspecies plant solid microbial inoculum, sterile normal saline and glucose according to the dosage ratio of 1g:6-8mL of. The compound prepared by the invention can improve blood lipid metabolism, can reduce The Cholesterol (TC) content and the Triglyceride (TG) content in serum and can increase the high density lipoprotein cholesterol (HDL-C) content.

Description

Compound for improving blood lipid metabolism and preparation method and application thereof
Technical Field
The invention relates to the field of health-care product methods, in particular to a compound for improving blood lipid metabolism and a preparation method and application thereof.
Background
Blood lipids are a general term for lipid components in blood. Lipids are a large group of neutral organic compounds that are insoluble in water and soluble in organic solvents (e.g., ethanol), and most commonly include cholesterol, triglycerides, phospholipids, and the like.
The lipid component in human blood can be obtained by processing in vivo after being absorbed from food, or can be synthesized in vivo by other substances. Lipid is an essential substance of human body, has very important physiological functions, and can bring adverse effects to the health if the lipid content in blood is too high or too low for some reason. Since the lipid components are insoluble in water and certainly not soluble in blood, it is necessary to combine the lipids with specific proteins to form so-called "lipoprotein" molecules, so as to circulate in the blood. In other words, blood lipids are actually present in the blood in the form of lipoproteins, which are complexes of lipids and proteins. Among them, the protein used for binding lipids is called "apolipoprotein".
The imbalance of blood lipid metabolism in a human body, the entrance and discharge of cholesterol and triglyceride, is called as dyslipidaemia, which is commonly called hyperlipidemia or hyperlipidemia. And even cause a variety of diseases, including liver tissue disorders such as the onset of nonalcoholic fatty liver disease (NAFLD).
Disclosure of Invention
In order to solve the technical problems, the invention provides a compound for improving blood lipid metabolism and a preparation method and application thereof, the compound prepared by fermenting radix curcumae, turmeric, chinese yam, hawthorn, giant knotweed and a microbial additive can improve blood lipid metabolism, can reduce the content of cholesterol (TC) and Triglyceride (TG) in serum, can regulate the content of high density lipoprotein cholesterol (HDL-C) in serum, and can regulate the levels of IL-6 and TNF-alpha in serum and the levels of TLR4 and NF-kappa B in liver tissues, thereby protecting liver cells.
The invention aims to provide a compound for improving blood lipid metabolism, which is prepared from the following components in parts by weight:
20-30 parts of radix curcumae, 10-12 parts of turmeric, 6-8 parts of Chinese yam, 4-6 parts of hawthorn, 4-6 parts of giant knotweed and 1-2 parts of microbial additive;
the microbial additive is prepared from a lactobacillus plantarum subspecies plantarum solid microbial inoculum, sterile normal saline and glucose according to the dosage ratio of 1g:6-8mL of.
Further, the microbial additive is prepared from a lactobacillus plantarum subspecies plantarum solid microbial inoculum, sterile normal saline and glucose according to the dosage ratio of 1g:6mL of: 0.3g of the above-mentioned raw materials.
Further, the preparation process of the lactobacillus plantarum subspecies plantarum solid microbial inoculum comprises the following steps: firstly, activating a strain Lactobacillus plantarum subspecies ATCC14917, transferring and purifying, selecting a single colony to inoculate into an MRS liquid culture medium, carrying out anaerobic culture for 46-48h at 100-120rpm and 35-37 ℃ to obtain a Lactobacillus plantarum subspecies ATCC14917 seed liquid, then inoculating the seed liquid into a fermentation tank according to the inoculation amount of 4-6% by volume for fermentation until the bacterial mass reaches 3-4 x 10 9 And (5) obtaining fermentation liquor by cfu/mL or more, and drying the fermentation liquor after wheat bran powder adsorption to obtain the lactobacillus plantarum subspecies plantarum solid microbial inoculum.
Further, the fermentation tank is 50L in specification, and the liquid filling amount in the fermentation tank is 20L.
Further, the fermentation conditions were 100rpm, 0.04MPa pot pressure, 37 ℃.
Further, the dosage ratio of the fermentation liquor to the wheat bran powder is 1L:2.5-3.5kg.
The second object of the present invention is to provide a method for preparing the above complex for improving blood lipid metabolism, comprising the steps of:
s1, weighing raw materials including radix curcumae, turmeric, chinese yam, hawthorn and giant knotweed in parts by weight, crushing the raw materials, uniformly mixing the raw materials, and sterilizing to obtain an aseptic composite raw material;
s2, preparing a microbial additive:
preparing a lactobacillus plantarum subspecies plantarum solid microbial inoculum, and then mixing the lactobacillus plantarum subspecies plantarum solid microbial inoculum with sterile normal saline and glucose according to the dosage ratio of 1g: uniformly mixing 0.3-0.4g of 6-8mL of the microbial additive;
s3, adding the microbial additive into the sterile composite raw material according to the parts by weight, carrying out anaerobic fermentation culture for 4-6 days at 37 ℃ and 38% humidity, drying the obtained fermentation product at 50 ℃ and obtaining the composite for improving blood lipid metabolism.
Further, the preparation process of the lactobacillus plantarum subspecies plantarum solid microbial inoculum comprises the following steps: firstly, activating strain Lactobacillus plantarum subspecies ATCC14917, transferring and purifying, selecting a single colony to inoculate into MRS liquid culture medium, anaerobically culturing at 100-120rpm and 35-37 ℃ for 46-48h to obtain Lactobacillus plantarum subspecies ATCC14917 seed liquid, then inoculating the seed liquid into a 50L fermentation tank according to the inoculation amount of 4-6% by volume for fermentation, wherein the liquid loading amount in the fermentation tank is 20L, and the fermentation is carried out at 100rpm, the tank pressure of 0.04MPa and the temperature of 37 ℃ until the bacterial mass reaches 3-4 × 10 9 Obtaining fermentation liquor with the dosage ratio of 1L:2.5-3.5kg of lactobacillus plantarum subspecies plant solid microbial inoculum is obtained by drying after wheat bran powder adsorption.
The third purpose of the invention is to provide the application of the compound in preparing the medicine for improving the blood lipid metabolism.
Further, the compound is used for preparing medicines for regulating and controlling the contents of cholesterol, triglyceride and high-density lipoprotein cholesterol in serum.
Compared with the prior art, the invention has the beneficial effects that:
1. the effects of the raw material components adopted in the invention are as follows:
the radix curcumae is used as a monarch drug in the invention, contains active substances such as curcumin, turmerone, tetrahydrocurcumin and the like, the tetrahydrocurcumin is a main bioactive metabolite of the curcumin, the radix curcumae is mainly used for protecting liver cells, promoting liver cell regeneration, removing fat and inhibiting liver cell fibrosis by reducing the levels of IL-6 and TNF-alpha and reducing the levels of TLR4 and NF-kB proteins in liver tissues, and improving liver lipid deposition and inflammatory reaction, and also has the effects of resisting oxidation, resisting tumors and resisting atherosclerosis;
turmeric is used as a ministerial drug to assist radix curcumae in improving liver lipid deposition and inflammatory reaction, and the principal components of turmeric comprise turmerone, arylturmerone, zingiberene, phellandrene, sabinene, eucalyptol, zedoarone, curcumenol, caryophyllene borneol, camphor and the like; mainly comprises curcumin and demethoxycurcumin; and bixin, norbixin, microelements and the like, the turmeric has the effects of activating blood and promoting qi, stimulating menstrual flow and relieving pain, can inhibit platelet aggregation, reduce blood plasma viscosity and whole blood viscosity, and also has the effects of protecting gastric mucosa and protecting liver cells;
the giant knotweed rhizome is used as an adjuvant drug for assisting the radix curcumae in improving blood lipid metabolism, reducing blood lipid and blood sugar, contains resveratrol and emodin, has the effects of reducing blood pressure, slowing down heart rate, strengthening myocardial contractility, protecting heart, inhibiting platelet aggregation, improving hemorheology, improving microcirculation, increasing white blood cells and platelets and resisting shock, and has the effects of stopping bleeding, resisting tumors, reducing blood sugar, reducing blood lipid, resisting inflammation and resisting oxidation.
The hawthorn is an adjuvant drug, contains triterpenes, bioflavonoids and rich vitamin C components, has the effects of expanding blood vessel walls, reducing cholesterol and triglyceride, reducing blood pressure and the like, and also contains maslinic acid and citric acid, and has remarkable blood fat reducing effect.
The yam is a messenger drug, not only coordinates the drugs, but also can obviously reduce the blood sugar, has the effects of preventing and treating diabetes, and can obviously resist the blood sugar rise caused by adrenalin and glucose;
according to the invention, through reasonable compatibility of the medicines and according to a specific compatibility principle of 'monarch, minister, assistant and guide', the raw materials are mutually cooperated and complemented, so that the blood fat metabolism is improved, the liver lipid deposition is improved, and the liver cells are effectively protected.
2. The compound prepared by the invention can be used for improving blood lipid metabolism, can reduce the content of cholesterol (TC) and Triglyceride (TG) in serum and can increase the content of high density lipoprotein cholesterol (HDL-C).
3. The compound prepared by the invention can protect liver cells by regulating the IL-6 and TNF-alpha levels in serum and the TLR4 and NF-kappa B protein levels in liver tissues.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. The experimental methods described in the examples of the present invention are all conventional methods unless otherwise specified.
Example 1
The embodiment provides a compound for improving blood lipid metabolism, which is prepared from the following components in parts by mass: 25g of radix curcumae, 11g of turmeric, 7g of Chinese yam, 5g of hawthorn, 5g of giant knotweed and 1.5g of microbial additive;
the microbial additive is prepared from a lactobacillus plantarum subspecies plantarum solid microbial inoculum, sterile normal saline and glucose according to the dosage ratio of 1g:6 mL;
the lactobacillus plantarum subspecies plant, the strain preservation number of which is ATCC14917, is purchased from Shanghai Fuxiang Biotechnology Limited;
the lactobacillus plantarum subspecies ATCC14917 is cultured by adopting an MRS culture medium, and the formula of the MRS solid culture medium is as follows: 10g of dried egg white, 5g of beef powder, 4g of yeast powder, 2g of glucose, 1ml of Tween 80, 2g of dipotassium phosphate, 5g of sodium acetate, 2g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate, 15g of agar powder and 1000ml of distilled water, wherein the pH value is 6.2. Removing agar powder from the corresponding MRS liquid culture medium.
The preparation process of the lactobacillus plantarum subspecies plantarum solid microbial inoculum comprises the following steps: firstly, inoculating a strain Lactobacillus plantarum subspecies ATCC14917 on an MRS solid culture medium for activation, transferring for 3-4 times to complete purification, selecting a single colony to inoculate into a triangular flask filled with a sterile MRS liquid culture medium, adding a latex plug into the triangular flask to create an anaerobic environment, culturing for 48 hours at 100rpm and 37 ℃ to obtain a Lactobacillus plantarum subspecies ATCC14917 seed solution, then inoculating the seed solution into a 50L fermentation tank according to the inoculation amount of 6% by volume ratio for fermentation culture, wherein the liquid loading amount in the fermentation tank is 20L, and fermenting at 100rpm, the tank pressure of 0.04Mpa and the temperature of 37 ℃ until the bacterial amount reaches 4 x 10 9 More than cfu/mL, adsorbing the obtained fermentation liquor by wheat bran powder of 50 meshes, wherein the dosage ratio of the fermentation liquor to the wheat bran powder is 1L:3.5kg, and vacuum drying at 50 ℃ to obtain the lactobacillus plantarum subspecies plantarum solid microbial inoculum.
The preparation method of the compound for improving blood lipid metabolism comprises the following steps:
s1, weighing the raw materials of turmeric root-tuber, turmeric, chinese yam, hawthorn and giant knotweed in parts by mass, crushing the raw materials, uniformly mixing the raw materials, and sterilizing to obtain an aseptic composite raw material;
s2, preparing the microbial additive according to the preparation method:
and S3, adding the microbial additive into the sterile composite raw material according to the mass ratio, carrying out anaerobic fermentation culture for 5 days at 37 ℃ and 38% humidity, drying the obtained fermentation product at 50 ℃, and obtaining the composite for improving blood lipid metabolism.
Example 2
Example 2 provides a complex for improving blood lipid metabolism, which is prepared substantially in the same manner as in example 1 except that:
the compound for improving blood lipid metabolism is prepared from the following components in parts by mass: 20g of radix curcumae, 10g of turmeric, 6g of Chinese yam, 4g of hawthorn, 4g of giant knotweed and 1g of microbial additive;
the microbial additive is prepared from a lactobacillus plantarum subspecies plantarum solid microbial inoculum, sterile normal saline and glucose according to the dosage ratio of 1g:7 mL;
in the step S3 of the preparation method of the compound, the microbial additive is added into the sterile compound raw material, the fermentation culture time is 4d, and the obtained fermentation product is dried at 55 ℃ to obtain the compound for improving the blood lipid metabolism.
Example 3
Example 3 provides a complex for improving blood lipid metabolism, which is prepared substantially in the same manner as in example 1 except that:
the compound for improving blood lipid metabolism is prepared from the following components in parts by mass: 30g of radix curcumae, 12g of turmeric, 8g of Chinese yam, 6g of hawthorn, 6g of giant knotweed and 2g of microbial additive;
the microbial additive is prepared from a lactobacillus plantarum subspecies plantarum solid microbial inoculum, sterile normal saline and glucose according to the dosage ratio of 1g:8mL of the raw materials are mixed into a mixture of 0.4 g;
in the step S3 of the preparation method of the compound, the microbial additive is added into the sterile compound raw material, the fermentation culture time is 6d, and the obtained fermentation product is dried at 60 ℃ to obtain the compound for improving the blood lipid metabolism.
1. Effect of the Complex for improving blood lipid metabolism prepared in example 1 of the present invention on blood lipid metabolism
1. The method comprises the following steps:
120 experimental mice with the weight of 22-25g are selected and randomly and evenly divided into a blank control group, a hyperlipemia model group, a bezafibrate control group (positive control), a high dose group, a medium dose group and a low dose group. Wherein the blank control group is fed with normal basal feed, and other groups except the basal feed are continuously drenched with the fat emulsion for 30 days, and the fat emulsion is drenched with the fat emulsion 12ml/kg every day to complete the model construction.
And (3) treatment intervention: the following treatments were given once daily:
the blank control group and the hyperlipidemia model group take physiological saline with the same amount, the bezafibrate control group takes 100mg/kg, the high dose group takes 25g/kg of the compound prepared by the application, the middle dose group takes 15g/kg of the compound prepared by the application, and the low dose group takes 5g/kg of the compound prepared by the application;
after the intervention for 10 days, the mouse eyeballs are picked and blood is taken, serum is separated, and The Cholesterol (TC) content, triglyceride (TG) content and high-density lipoprotein cholesterol (HDL-C) content in the serum are measured. The results are reported in table 1.
Preparing the fat emulsion:
adding 40g lard into 250ml water, heating to melt, adding 20g cholesterol to melt, adding 4g sodium cholate and 2g propyl thiouracil tablet, stirring, adding appropriate amount of distilled water and 40ml each of polysorbate-80 (Tween 80) and 1,2 propylene glycol, stirring, adding double distilled water to 200ml, and mixing to obtain fat emulsion.
TABLE 1 Effect of the complexes prepared according to the invention on the metabolism of blood lipids
Grouping TC(mmol/L) TG(mmol/L) HDL-C(mmol/L)
Blank control group 3.26±0.22 1.54±0.26 1.77±0.25
Hyperlipemia model group 4.03±0.14 2.03±0.31 0.98±0.12
Positive control group 3.61±0.19 1.45±0.22 1.24±0.15
High dose group 3.08±0.38 1.52±0.26 1.76±0.24
Middle dose group 3.15±0.35 1.54±0.27 1.69±0.20
Low dose group 3.55±0.28 1.58±0.29 1.61±0.17
As can be seen from Table 1, the compound prepared by the invention can effectively reduce The Cholesterol (TC) content and the Triglyceride (TG) content in serum, and can effectively improve the high-density lipoprotein cholesterol (HDL-C) content in serum. Compared with bezafibrate medicaments in a positive control group, the compound prepared by the invention has obviously better adjusting effect on the contents of cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) in serum than the positive control, and only slightly inferior to the positive control in the adjustment on the content of Triglyceride (TG).
It should be noted that the compound provided by the invention is not suitable for pregnant women.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Claims (10)

1. The compound for improving blood lipid metabolism is characterized by being prepared from the following components in parts by weight: 20-30 parts of radix curcumae, 10-12 parts of turmeric, 6-8 parts of Chinese yam, 4-6 parts of hawthorn, 4-6 parts of giant knotweed and 1-2 parts of microbial additive;
the microbial additive is prepared from a lactobacillus plantarum subspecies plantarum solid microbial inoculum, sterile normal saline and glucose according to the dosage ratio of 1g:6-8mL of;
the compound for improving blood lipid metabolism is prepared by the following steps:
s1, weighing raw materials including radix curcumae, turmeric, chinese yam, hawthorn and giant knotweed in parts by weight, crushing the raw materials, uniformly mixing the raw materials, and sterilizing to obtain an aseptic composite raw material;
s2, preparing a microbial additive:
preparing a lactobacillus plantarum subspecies vegetabilis solid microbial inoculum, and then mixing the lactobacillus plantarum subspecies vegetabilis solid microbial inoculum with sterile normal saline and glucose according to the dosage ratio of 1g: uniformly mixing 0.3-0.4g of 6-8mL of the microbial additive;
and S3, adding the microbial additive into the sterile composite raw material according to the parts by weight, carrying out anaerobic fermentation culture for 4-6 days at 37 ℃ and 38% humidity, drying the obtained fermentation product at 50-60 ℃ to obtain the compound for improving blood lipid metabolism.
2. The composition for improving blood lipid metabolism according to claim 1, wherein the microbial additive is prepared from lactobacillus plantarum subspecies plantarum solid bacterial agent, sterile physiological saline and glucose in an amount ratio of 1g:6mL of: 0.3g of the above-mentioned raw materials.
3. The compound for improving blood lipid metabolism according to claim 2, wherein the lactobacillus plantarum subspecies plantarum solid bacterial agent is prepared by the following process: firstly, activating a strain Lactobacillus plantarum subspecies ATCC14917, transferring and purifying, selecting a single colony to inoculate into an MRS liquid culture medium, carrying out anaerobic culture for 46-48h at 100-120rpm and 35-37 ℃ to obtain a Lactobacillus plantarum subspecies ATCC14917 seed liquid, then inoculating the seed liquid into a fermentation tank according to the inoculation amount of 4-6% by volume for fermentation until the bacterial mass reaches 3-4 x 10 9 And (5) obtaining fermentation liquor by cfu/mL or more, and drying the fermentation liquor after wheat bran powder adsorption to obtain the lactobacillus plantarum subspecies plantarum solid microbial inoculum.
4. The composition for improving blood lipid metabolism according to claim 3, wherein the fermenter has a 50L size, and the amount of liquid in the fermenter is 20L.
5. The composition for improving blood lipid metabolism according to claim 4, wherein the fermentation conditions are 100rpm, 0.04MPa in pot pressure and 37 ℃.
6. The complex for improving blood lipid metabolism according to claim 5, wherein the ratio of the fermentation broth to wheat bran powder is 1L:2.5-3.5kg.
7. A method for preparing a complex for improving blood lipid metabolism according to any one of claims 1 to 6, comprising the steps of:
s1, weighing raw materials including radix curcumae, turmeric, chinese yam, hawthorn and giant knotweed in parts by weight, crushing the raw materials, uniformly mixing the raw materials, and sterilizing to obtain an aseptic composite raw material;
s2, preparing a microbial additive:
preparing a lactobacillus plantarum subspecies plantarum solid microbial inoculum, and then mixing the lactobacillus plantarum subspecies plantarum solid microbial inoculum with sterile normal saline and glucose according to the dosage ratio of 1g: uniformly mixing 0.3-0.4g of 6-8mL of the microbial additive;
and S3, adding the microbial additive into the sterile composite raw material according to the parts by weight, carrying out anaerobic fermentation culture for 4-6 days at 37 ℃ and 38% humidity, drying the obtained fermentation product at 50-60 ℃ to obtain the compound for improving blood lipid metabolism.
8. The method for preparing a complex for improving blood lipid metabolism according to claim 7, wherein the lactobacillus plantarum subspecies plantarum solid bacterial agent is prepared by the following steps: firstly, activating a strain Lactobacillus plantarum subspecies ATCC14917, transferring and purifying, selecting a single colony, inoculating the single colony into an MRS liquid culture medium, carrying out anaerobic culture for 46-48h at 100-120rpm and 35-37 ℃ to obtain a Lactobacillus plantarum subspecies ATCC14917 seed solution, and then inoculating the seed solution into a 50L fermentation tank according to the volume ratio of 4-6 percent of the inoculation amountFermenting at a fermentation tank with liquid loading of 20L under a pressure of 0.04MPa at 100rpm and 37 deg.C until the strain amount reaches 3-4 × 10 9 Obtaining fermentation liquor with the dosage ratio of 1L:2.5-3.5kg of lactobacillus plantarum subspecies plant solid microbial inoculum is obtained by drying after wheat bran powder adsorption.
9. Use of a complex according to any one of claims 1 to 6 in the manufacture of a medicament for improving blood lipid metabolism.
10. Use of a complex according to claim 9 for the preparation of a medicament for improving blood lipid metabolism, wherein the complex is used for the preparation of a medicament for modulating the cholesterol, triglyceride and high density lipoprotein cholesterol levels in serum.
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