CN-121975035-A - Preparation method of fructus alpiniae oxyphyllae pectin polysaccharide and application of fructus alpiniae oxyphyllae pectin polysaccharide in treating Alzheimer disease

Abstract

The invention discloses a preparation method of fructus alpiniae oxyphyllae pectin polysaccharide and application thereof in treating Alzheimer's disease. The pectic polysaccharide is prepared from fructus alpiniae oxyphyllae serving as a raw material through the steps of degreasing, water extraction, alcohol precipitation, deproteinization, dialysis, chromatographic purification and the like, and is uniform acidic polysaccharide. The structural characterization result shows that the pectic polysaccharide mainly comprises rhamnose, glucuronic acid, galacturonic acid and galactose, wherein the content of galacturonic acid is dominant, and partial residues are subjected to O-methylation and/or O-acetylation modification, so that the pectic polysaccharide has a pectin-type acidic polysaccharide structure with alpha- (1- & gt 4) -D-galacturonic acid as a main chain and branched chains. Animal experiment results show that the fructus alpiniae oxyphyllae pectin polysaccharide can remarkably improve learning and memory capacity and cognitive dysfunction of Alzheimer disease model animals. The fructus alpiniae oxyphyllae pectin polysaccharide provided by the invention has definite sources and stable preparation method, can be used for preparing a pharmaceutical composition or functional food for preventing and treating Alzheimer disease, and has good application prospect.

Inventors

• CHEN XINGLONG
• Hu mingzhu
• ZHANG RONGPING
• NI GUANGHUI
• HU WEIYAN
• LI XINGXI
• DONG JUNFANG
• LI HUAN

Assignees

• 云南中医药大学

Dates

Publication Date

20260505

Application Date

20260302

Claims (10)

1. 1. A fructus alpiniae oxyphyllae pectin polysaccharide, which is characterized in that monosaccharide composition of the fructus alpiniae oxyphyllae pectin polysaccharide comprises rhamnose, glucuronic acid, galacturonic acid and galactose.
2. 2. The alpinia oxyphylla pectin polysaccharide according to claim 1, wherein the monosaccharide composition of the alpinia oxyphylla pectin polysaccharide comprises 4.0% rhamnose, 9.8% glucuronic acid, 72.8% galacturonic acid and 1.6% galactose in mole percent.
3. 3. The fructus alpiniae oxyphyllae pectin polysaccharide of claim 1, wherein the total sugar content of the fructus alpiniae oxyphyllae pectin polysaccharide is 91.8% and the total uronic acid content is 75.2%.
4. 4. The fructus alpiniae oxyphyllae pectin polysaccharide of claim 1, wherein the fructus alpiniae oxyphyllae pectin polysaccharide has a relative molecular weight of 19.35 kDa.
5. 5. The fructus alpiniae oxyphyllae pectin polysaccharide according to any one of claims 1 to 4, wherein the fructus alpiniae oxyphyllae pectin polysaccharide is pectin-type acidic polysaccharide, alpha- (1- > 4) -D-galacturonic acid residues are used as a main chain structure, and part of galacturonic acid residues in the main chain are subjected to O-methylation and/or O-acetylation modification, and the fructus alpiniae oxyphyllae pectin polysaccharide contains a branched chain structure formed by beta- (1- > 6) -D-glucuronic acid, alpha-L-rhamnose and beta-D-galactose residues.
6. 6. The method for preparing the fructus alpiniae oxyphyllae pectin polysaccharide as set forth in any one of claims 1 to 5, wherein the method comprises the following steps: (1) Preprocessing, namely crushing fructus alpiniae oxyphyllae, extracting by adopting methanol under reflux, and drying dregs after the reflux is finished to obtain fructus alpiniae oxyphyllae powder, wherein the feed liquid ratio of the fructus alpiniae oxyphyllae to the methanol is 1:5-1:10; (2) Repeatedly boiling and extracting the fructus alpiniae oxyphyllae powder obtained in the step (1) by using boiling water, wherein the feed liquid ratio of the fructus alpiniae oxyphyllae powder to the water is 1:5-1:10, collecting an extracting solution, and carrying out suction filtration and concentration to obtain a fructus alpiniae oxyphyllae polysaccharide crude extracting solution; (3) The preparation method comprises the steps of (1) dispersing a crude extract of the fructus alpiniae oxyphyllae polysaccharide obtained in the step (2) with water, adding ethanol with the volume fraction of 60-100% for alcohol precipitation, carrying out secondary alcohol precipitation to remove part of alcohol-soluble impurities, then placing the obtained product in a dialysis bag with the molecular weight cut-off range of 1000-6000 Da for dialysis for 12-72 hours to obtain a dialysis solution, adding a Sevage reagent into the dialysis solution for deproteinization treatment, wherein the Sevage reagent is a mixed solution of chloroform and n-butanol, the volume ratio of the chloroform to the n-butanol is 1:1-8:1, centrifuging, taking a supernatant, concentrating, carrying out suction filtration and freeze-drying to obtain the refined fructus alpiniae oxyphyllae pectin polysaccharide.
7. 7. The method according to claim 6, wherein the ethanol in the ethanol precipitation step has a volume fraction of 70-95%.
8. 8. The use of a alpinia oxyphylla pectin polysaccharide according to any one of claims 1 to 5 for preparing a medicament for preventing and/or treating alzheimer's disease.
9. 9. The use according to claim 8, wherein the alpinia oxyphylla pectin polysaccharide is used for improving cognitive dysfunction and/or learning memory decline associated with alzheimer's disease.
10. 10. The use according to claim 8 or 9, wherein the alpinia oxyphylla pectin polysaccharide is formulated as an oral preparation or a functional food.

Description

Preparation method of fructus alpiniae oxyphyllae pectin polysaccharide and application of fructus alpiniae oxyphyllae pectin polysaccharide in treating Alzheimer disease Technical Field The invention belongs to the technical field of development and utilization of natural products, and particularly relates to a preparation method and structural characterization of pectic polysaccharide from fructus alpiniae oxyphyllae and application of pectic polysaccharide in preventing and treating Alzheimer disease. Background Alzheimer's Disease (AD) is a common neurodegenerative Disease whose main pathological features include senile plaques formed by beta-amyloid (Abeta) deposition, neurofibrillary tangles formed by Tau protein hyperphosphorylation, and neuronal loss. According to 2021, china's Alzheimer's disease report, by 2019, AD patients in China are more than 1000 ten thousand, and the characteristics of high morbidity and high mortality are presented. Currently only two classes of drugs, cholinesterase inhibitors and N-methyl-D-aspartate antagonists, are clinically approved for the treatment of AD, but these drugs only control symptoms and cannot alter disease progression. Therefore, the development of the novel drug for preventing and treating the Alzheimer disease, which has high safety, is suitable for long-term use and has various action mechanisms, has important significance. Fructus Alpinae Oxyphyllae is dried mature fruit of Alpinae Oxyphyllae (Alpinia oxyphylla Miq.) of Zingiberaceae, and has effects of warming spleen, relieving diarrhea, warming kidney, and stopping nocturnal emission. Modern researches show that the fructus alpiniae oxyphyllae contains rich polysaccharide components and has a neuroprotective effect, but the application of the fructus alpiniae oxyphyllae in the aspect of AD prevention and treatment induced by D-galactose has not been systematically researched and reported. Compared with the traditional chemical medicines, the alpinia oxyphylla polysaccharide has the characteristics of multi-target and multi-path effects, can simultaneously interfere a plurality of pathological links of AD, has higher safety, and is more suitable for long-term use. However, the existing researches focus on crude fructus alpiniae oxyphyllae polysaccharide, and the specific structural type and the role of the crude polysaccharide in Alzheimer disease are not systematically researched. Pectin is an acidic polysaccharide that is widely found in the plant kingdom, mainly in the roots, stems, leaves and fruits of higher plants. Pectin is approved by the joint Committee of food additives of the Shiwei organization as a food additive, has no daily dose limitation, and has high safety. In addition, pectin has anti-inflammatory and hypolipidemic physiological effects due to its antioxidant activity. It also promotes digestive health and may be a prebiotic. These functions of pectin benefit from its unique structure. Its composition generally includes four kinds of homogalacturonic acid (HG), rhamnogalacturonan I (RG-I), rhamnogalacturonan II (RG-II) and xylogalacturonic acid (XGA), and various branched side chains. In particular, pectins can be classified by the degree of methyl esterification (DM) into high methoxy pectins and low methoxy pectins. Pectic polysaccharides are widely used because of their softness, flexibility, high water content, natural compatibility and unique structure. They are commonly used in food packaging, probiotic drug delivery systems and as satiety inducing agents for the relief of obesity. However, there is no report on systematic study of fructus alpiniae oxyphyllae-derived pectin polysaccharide. Disclosure of Invention The invention aims to provide a uniform fructus alpiniae oxyphyllae pectic polysaccharide and a preparation method thereof, and further provides application of the pectic polysaccharide in preventing and treating Alzheimer disease. The monosaccharide composition of fructus Alpinae Oxyphyllae pectin polysaccharide comprises rhamnose, glucuronic acid, galacturonic acid and galactose. The monosaccharide component of the fructus alpiniae oxyphyllae pectin polysaccharide comprises 4.0% of rhamnose, 9.8% of glucuronic acid, 72.8% of galacturonic acid and 1.6% of galactose in mole percent. The total sugar content of fructus Alpinae Oxyphyllae pectin polysaccharide is 91.8%, and the total uronic acid content is 75.2%. The relative molecular weight of the fructus alpiniae oxyphyllae pectin polysaccharide is 19.35 kDa. The fructus alpiniae oxyphyllae pectin polysaccharide is pectin acid polysaccharide, alpha- (1- & gt 4) -D-galacturonic acid residue is taken as a main chain structure, O-methylation and/or O-acetylation modification is carried out on part of galacturonic acid residues in the main chain, and the fructus alpiniae oxyphyllae pectin polysaccharide contains a branched chain structure formed by beta- (1- & gt 6) -D-glucuronic acid, alpha-L-rhamnose and beta-D-galact