阅读说明：本技术 唾液酸在调节由引起阿兹海默病的基因的异常表达造成的血脂代谢异常中的应用 (Use of sialic acid for modulating dyslipidemias caused by abnormal expression of genes responsible for alzheimer's disease ) 是由 汪志明 李翔宇 丰珂珏 肖敏 陆姝欢 于 2020-05-08 设计创作，主要内容包括：本发明涉及唾液酸在调节由引起阿兹海默病的基因的异常表达造成的血脂代谢异常中的应用,具体为调节由APP和PS1基因表达水平异常导致的血脂代谢异常中的应用。本发明经过研究发现,对APP/PS1转基因小鼠施用唾液酸,发现在补充唾液酸后,其载脂蛋白A与载脂蛋白B表达水平均有改善,说明了唾液酸可以有效调节APP和PS1基因表达异常造成的血脂代谢异常。本发明将唾液酸作为饮食干预的方式或制备成药物制剂对血脂代谢异常进行调节,具有来源丰富、毒副作用小、对人群适用性广等优势,除了单一使用,还能够灵活组方,在由APP和PS1基因表达异常引起的血脂代谢异常的调节中具有广阔的应用前景。(The invention relates to application of sialic acid in regulating dyslipidemia caused by abnormal expression of genes causing Alzheimer's disease, in particular to application in regulating dyslipidemia caused by abnormal expression levels of APP and PS1 genes. According to research, the application of sialic acid to APP/PS1 transgenic mice shows that after sialic acid is supplemented, the expression levels of apolipoprotein A and apolipoprotein B are improved, and the fact that sialic acid can effectively regulate dyslipidemia caused by abnormal expression of APP and PS1 genes is shown. The sialic acid is used as a dietary intervention mode or prepared into a medicinal preparation for regulating dyslipidemia, has the advantages of rich sources, small toxic and side effects, wide applicability to people and the like, can be flexibly prepared in addition to single use, and has wide application prospect in regulation of dyslipidemia caused by abnormal expression of APP and PS1 genes.)

1. Use of sialic acid to modulate dyslipidemias caused by abnormal expression of genes responsible for alzheimer's disease.

2. The use according to claim 1, wherein the genes responsible for alzheimer's disease are APP and PS 1.

3. The use according to claim 1 or 2, wherein the dyslipidemia is an abnormality of apolipoprotein a and apolipoprotein B in the blood.

4. Use according to any one of claims 1 to 3, wherein the sialic acid is free sialic acid or in bound form; the sialic acid in the conjugated form is obtained by combining sialic acid with any one or more of oligosaccharide conjugate, lipid conjugate and protein conjugate; the sialic acid is preferably present in the form of N-acetylneuraminic acid.

5. Use according to any one of claims 1 to 4, wherein the sialic acid, when used, is administered daily at 1 to 50mg/Kg relative to the body weight of the human, preferably daily at 1 to 35mg/Kg relative to the body weight of the human.

6. Use according to claim 5, wherein the sialic acid is administered at the time of use in the form of one or more of a nutritional supplement, a food, a beverage or a pharmaceutical preparation.

7. The use as claimed in claim 6, wherein the dosage forms of the pharmaceutical formulation and the nutritional supplement include, but are not limited to, one or more of tablets, powders, granules, capsules, injections, sprays, films, suppositories, oral liquids, nasal drops or drop pills.

8. Use according to any one of claims 1 to 7, wherein the sialic acid is supplemented in use with one or more of DHA, EPA, alpha-linolenic acid, lecithin, rutin, quercetin, phytosterols, chitosan, nuciferine, green tea polyphenols, laminarin, lentinan.

9. A formulation for modulating dyslipidemia due to abnormal expression of genes causing alzheimer's disease; the preparation takes sialic acid as an active ingredient.

10. The preparation according to claim 9, wherein the sialic acid is present in an amount of 0.01 to 100% by weight of the preparation.

Technical Field

The invention relates to the field of biotechnology, in particular to application of sialic acid in regulating dyslipidemia caused by abnormal expression of genes causing Alzheimer disease.

Background

Alzheimer's Disease (AD) is a chronic degenerative disease of the central nervous system, commonly known as senile dementia, the most common form of senile dementia, and is characterized by a progressive worsening of the disease condition, characterized by memory impairment and cognitive dysfunction. In the elderly people over 60 years old, the disease is more likely to occur, and the prevalence rate is increased rapidly as the population becomes older. The disease condition of AD can continue for 20 years, the disease condition gradually worsens, the independent living ability is gradually lost, and the AD dies due to complications 10-20 years after the disease occurs, so that heavy burden and pain are brought to individuals, families and the society.

The pathogenesis of the alzheimer disease is not completely defined, and the main pathogenesis of the alzheimer disease is amyloid beta theory, tau protein theory, gene mutation theory, cholinergic theory and the like, wherein the scholars generally consider that the cytotoxic amyloid beta (a β) plays an important role in the pathogenesis of the AD, and the a β is the chief culprit of the AD. A β is produced from β Amyloid Precursor Protein (APP) by cleavage with β -secretase and γ -secretase. The up-regulation of APP, which acts as a donor for a β production, directly increases a β production and contributes to the pathogenesis of AD. While presenilin 1(PS1) is the main functional component of gamma-secretase, the functional abnormality of presenilin 1 can affect the activity of gamma-secretase, and many evidences indicate that PS1 plays an important role in the activity of gamma-secretase and the generation of A beta, and also suggest that the regulation and control of the expression of PS1 gene can be directly connected with the occurrence and development of AD. Therefore, in recent years, it has been considered that AD patients may have over-expression of APP and PS1 genes. The animal models in the past researches can not well simulate the progressive process of human AD diseases, but the problem is solved by the appearance of an APP/PS1 double-transgenic mouse model, the APP/PS1 double-transgenic mouse model has two gene mutations, namely the mutation of an APP gene and the deletion of a No. 9 exon of a PS1 gene, and the mouse model can better show early mild cognitive impairment, which is also more typical behavior and pathological morphological change in the AD process. And the APP/PS1 double-transgenic mice of low-age form Abeta pathological plaques, so that in-vivo tests of medicaments can be performed, the test time is greatly shortened, the test period of researchers is accelerated, and the application prospect in the research of evaluating and treating the Alzheimer's disease by the medicaments is wide.

Sialic Acid (SA) is a generic term for a class of derivatives having the basic structure of the nine-carbon sugar neuraminic acid, of which N-acetylneuraminic acid is the most predominant member. Sialic acid is widely found in cell membrane glycoproteins and lipoproteins in organisms and plays an important role in many important processes in organisms. Along with the attention and research on the biochemical characteristics and biological activity of sialic acid, the sialic acid has become widely applied in various fields such as food, daily chemical products, medicines and the like. The existing research shows that the sialic acid has a plurality of physiological functions, such as improving the intelligence and memory of infants, resisting senile dementia, whitening, resisting aging, improving the immunity of human bodies, resisting bacteria and viruses, improving the absorption of vitamins and mineral substances by intestinal tracts and the like.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides application of sialic acid in regulating abnormal blood lipid metabolism caused by abnormal expression of genes causing Alzheimer disease. In particular to Alzheimer's disease associated with abnormal expression of two genes of APP and PS1 and the caused abnormality of apolipoprotein A and apolipoprotein B in blood fat.

The sialic acid regulation effect of the invention can specifically improve the apolipoprotein A expression level of the Alzheimer disease patient related to the abnormal expression of APP and PS1 genes and reduce the apolipoprotein B expression level of the patient.

The apolipoprotein belongs to the protein part in plasma lipoprotein, and clinical research shows that the apolipoprotein can convey blood fat required by an organism to each tissue of the body, promote normal metabolism of the organism and reasonably utilize protein. However, when apolipoprotein is abnormal, it will block the metabolism and utilization of blood lipid, and thus it is involved in the occurrence, development and prognosis of various diseases, such as hypertension, atherosclerosis, hyperlipidemia, etc.

Apolipoprotein ApoA1 is the major apolipoprotein for HDL-C, and can evaluate HDL-C level, combine free cholesterol in peripheral tissues, promote cholesterol clearance in arterial wall cells, and accelerate cholesterol metabolism in liver, and clinical data show that HDL and ApoA1 levels are significantly lower in patients with cardiovascular and cerebrovascular diseases than in normal persons. Apolipoprotein ApoB is an apolipoprotein of arteriosclerosis-actuating lipoprotein such as VLDL-C, LDL-C and the like, is in positive correlation with LDL-C level, and clinical data show that the blood lipid level and the ApoB level of a patient with cardiovascular and cerebrovascular diseases are obviously higher than those of a normal person.

The invention unexpectedly discovers in a project for researching the influence of sialic acid on Alzheimer's disease that except that the expression of brain Abeta peptide is increased and deposition occurs in APP/PS1 mice used for constructing a model, apolipoprotein A and apolipoprotein B in blood of the APP/PS1 mice are obviously different from normal mice under the condition of normal feeding, the apolipoprotein A level is obviously lower, the apolipoprotein B level is obviously higher than that of the normal mice, and the phenomena show that the lipid metabolism of the APP/PS1 mice is greatly abnormal.

The research results also show that: under the same condition, the ApoA1 level of the mice fed with SA is obviously increased and is higher than or close to the level of normal mice, and the effects generated by different SA doses are not obviously different; thus, the low dosage of SA can play a role in regulating blood lipid metabolism. While mice fed with SA had significantly lower ApoB levels than AD mice, and mice fed with high doses of SA had the best results, with ApoB levels comparable to normal mice. The above phenomena can indicate that SA has a positive improvement effect on the dyslipidemia phenomenon caused by the Alzheimer disease associated with abnormal expression of the APP and PS1 genes.

Reference to "sialic acid" in the present application is a reference to any single member of the 9-carbon sugar neuraminic acid derivative family known in the art, which includes sialic acid in bound form as well as free sialic acid. For example monomeric N-acetylneuraminic acid or polysialic acid; sialic acids bound to proteins (such as casein glycomacropeptide or mucin); sialic acid bonded to a carbohydrate (e.g. sialyllactose or a bacterial capsular polysaccharide of sialic acid); and sialic acids bound to lipids (such as gangliosides).

Sialic acid as described in the present invention is preferably present in the form of N-acetylneuraminic acid.

The invention further provides the use of sialic acid in the manufacture of a nutritional supplement, food, beverage or pharmaceutical formulation for modulating dyslipidemias caused by abnormal expression of genes responsible for alzheimer's disease.

Further, when applied to food, it may be various kinds of candies, cookies, pastries, dairy products, etc.

Further, when applied to a pharmaceutical preparation or a nutritional supplement, the dosage form may not be limited to one or more of tablets, powders, granules, capsules, injections, sprays, films, suppositories, oral liquids, nasal drops or dropping pills.

In some embodiments, the dosage form is administered to a patient with alzheimer's disease at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day, at least six times a day, at least seven times a day, at least eight times a day, at least nine times a day, at least ten times a day.

In some embodiments sialic acid is administered daily at (1-50) mg/Kg relative to body weight, preferably sialic acid is administered daily at (1-35) mg/Kg relative to body weight.

In some embodiments, the administration is continuous for four months or more to the alzheimer's patient.

In some embodiments, the administration is supplemented with one or more of DHA, EPA, alpha-linolenic acid, lecithin, rutin, quercetin, phytosterols, chitosan, nuciferine, green tea polyphenols, laminarin, and lentinan at the same time.

In some embodiments, the sialic acid comprises 0.01% to 100% by weight of the pharmaceutical preparation or nutritional supplement. In some more specific embodiments, the crystalline sialic acid powder can be directly used for tabletting or granule preparation without adding other auxiliary materials after granulation.

The invention has the following beneficial effects:

the mouse with abnormal APP and PS1 gene expression is a common AD model mouse, and the research of the invention finds that the blood lipid metabolism of the mouse is abnormal and can be adjusted by supplementing sialic acid. Animal experiments prove that after sialic acid is supplemented to an AD model mouse, the average expression level of apolipoprotein A and apolipoprotein B tends to be normal, which shows that sialic acid can effectively regulate the abnormal levels of apolipoprotein A and apolipoprotein B caused by the abnormal expression of APP and PS1 genes, promote the blood metabolism level to be normal and possibly further influence cardiovascular and cerebrovascular diseases caused by Alzheimer's disease. Therefore, the sialic acid is supplemented as a mode of dietary intervention or prepared into a medicinal preparation, the preparation has the advantages of rich sources, small toxic and side effects, wide applicability to human groups and the like, can be flexibly prepared besides being singly used, and has wide application prospect in regulation of dyslipidemia caused by abnormal expression of APP and PS1 genes.

Drawings

FIG. 1 is a diagram of APP/PS1 qPCR in 2 × Tg-AD experimental mice provided in example 1 of the present invention;

FIG. 2 is a graph showing the change in the expression level of apolipoprotein A1(ApoA1) in 2 × Tg (APP/PS1) -AD male mice and blank control male mice provided in example 1 of the present invention;

FIG. 3 is a graph showing the change in the expression level of apolipoprotein B (ApoB) in 2 × Tg (APP/PS1) -AD male mice and a blank control male mouse provided in example 1 of the present invention;

FIG. 4 is a graph showing the change in the expression levels of apolipoprotein A1(ApoA 1)/lipoprotein B (ApoB) in 2 × Tg (APP/PS1) -AD mice and blank control male mice provided in example 1 of the present invention.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

The sialic acid of the invention is derived from a sialic acid product produced by the applicant, and the purity of the N-acetylneuraminic acid is more than or equal to 98 percent.

Example 1

1. Test article

Sialic acid low dose: 17mg/kg/day body weight, measured in sialic acid: 84mg/kg/day body weight, sialic acid high dose: 420mg/kg/day body weight.

2. Sample set-up

The experiment adopts a double-transgenic model of APP/PS1, the model is a common AD mouse model with two genes of APP/PS1 transferred to a mouse, and A beta can be generated in the brain in a large amount.

Male 2 × Tg (APP/PS1) -AD mice (hereinafter AD mice) were grouped with placebo male mice according to the following table. Where L is the low dose, M is the medium dose, and H is the high dose (WT is a normal mouse).

TABLE 1 grouping of AD Male mice with placebo Male mice

3. Experimental procedure

(1) The test substance and placebo were mixed into the feed for mice, respectively, and the mice were treated with free access to food, with the feed being changed daily.

(2) The blank control group and the test model group were given commercial experimental mouse feed. After 1 month of feeding, the blank group and the model group were administered with placebo or test substance according to the experimental design under the condition that the feed was not changed, and were continuously administered for 4 months to 6 months.

(3) The blood lipid status and apolipoprotein level of each group of mice are detected.

4. Detecting the index

(1) Apolipoprotein a1(ApoA1) levels.

(2) Apolipoprotein b (apob) level.

5. The result of the detection

FIG. 1 is a diagram of APP/PS1 qPCR in 2 × Tg-AD experimental mice provided in this example.

The results of the level measurements for each group of apolipoprotein a1(ApoA1) and apolipoprotein b (apob) are shown in fig. 2, 3, and 4.

The apolipoprotein belongs to the protein part in plasma lipoprotein, and clinical research shows that the apolipoprotein can convey blood fat required by an organism to each tissue of the body, promote normal metabolism of the organism and reasonably utilize protein. However, when apolipoprotein is abnormal, it will block the metabolism and utilization of blood lipid, and thus it is involved in the occurrence, development and prognosis of various diseases, such as hypertension, atherosclerosis, hyperlipidemia, etc.

Apolipoprotein ApoA1 is the major apolipoprotein for HDL-C, and can evaluate HDL-C level, combine free cholesterol in peripheral tissues, promote cholesterol clearance in arterial wall cells, and accelerate cholesterol metabolism in liver, and clinical data show that HDL and ApoA1 levels are significantly lower in patients with cardiovascular and cerebrovascular diseases than in normal persons. In the research, compared with the AD mice with abnormal blood fat, the level of ApoA1 of the mice fed with SA is obviously increased, and is higher than or close to the level of normal mice, and the effects generated by different SA doses are not obviously different, which shows that the low-dose SA can play a role in regulating blood fat.

Apolipoprotein ApoB is an apolipoprotein of arteriosclerosis-actuating lipoprotein such as VLDL-C, LDL-C and the like, is in positive correlation with LDL-C level, and clinical data show that the blood lipid level and the ApoB level of a patient with cardiovascular and cerebrovascular diseases are obviously higher than those of a normal person. In the research, the ApoB level of the dyslipidemia AD mice is obviously higher than that of the normal mice, the ApoB of the mice fed with SA is obviously reduced compared with the AD mice, the effect of the mice fed with SA at a high dose is the best, and the ApoB level of the mice fed with SA is equivalent to that of the normal mice.

It can also be seen from the ApoA1/ApoB values in fig. 4 that the ratio of SA-fed mice was significantly different from AD mice, approaching the ApoA1/ApoB values of normal mice.

6. Summary of the invention

In conclusion, when the sialic acid is fed to the mice, the ApoA1 and ApoB indexes are obviously influenced, and the ApoA1 level and the ApoB level of the dyslipidemia mice can be obviously increased and decreased. Therefore, the sialic acid has a certain positive effect on regulating blood fat and preventing cardiovascular and cerebrovascular diseases.

Conversion from mouse to Adult dosages to (1-35) mg/Kg/day was performed according to the FDA recommended guidelines, estimation, the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in addition health volumes. The dosage can be converted according to the weight of an adult, for example, the dosage is calculated according to the weight of 60kg of the adult, and the daily intake of sialic acid after conversion is respectively as follows: low dose 82.62mg, medium dose 408.24mg, high dose 2041.2 mg.

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.