Maca polysaccharide of immunoregulatory activity and preparation method therefor

A kind of technology of polysaccharide and maca dry tablets, which is applied in the field of medicine

Inactive Publication Date: 2016-03-02
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on the non-starch polysaccharides in Maca at home and abroad. Our laboratory has carried out research on this aspect, and conducted a preliminary discussion on its structure, composition and activity. It is found that the structure of Maca polysaccharid

Method used

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  • Maca polysaccharide of immunoregulatory activity and preparation method therefor
  • Maca polysaccharide of immunoregulatory activity and preparation method therefor
  • Maca polysaccharide of immunoregulatory activity and preparation method therefor

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Experimental program
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Effect test

Embodiment 1

[0015] The preparation process route of Maca polysaccharide:

[0016] (1) Dry Maca tablets are degreased with 80-95% ethanol under reflux, water is added to make the ratio of solid to liquid 1:15-25, and boiling water is refluxed for extraction. The reaction time is 1.5 h.

[0017] (2) Precipitate the extracted supernatant with ethanol with a final concentration of 70-90%, let it stand overnight at 4° C., and collect the precipitate by centrifugation.

[0018] (3) Redissolve with double distilled water at a ratio of 1:15 to 25, stir at 60°C for 4 hours, collect the supernatant by centrifugation, precipitate the supernatant with ethanol with a final concentration of 70% to 90%, and let stand at 4°C Overnight, the precipitate was collected by centrifugation to obtain crude polysaccharide.

[0019] (4) Add the above-mentioned crude polysaccharide to double-distilled water to make a solution with a concentration of 15-25 mg / mL, and centrifuge to remove the supernatant for later u...

Embodiment 2

[0022] Structural Identification of Maca Polysaccharide

[0023] (1) Pre-column derivatization method to detect its monosaccharide composition

[0024] figure 1 It is the standard spectrum of nine kinds of monosaccharides. As shown in the figure, the order of the peak times of the nine kinds of monosaccharide standards is mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, and galactose , arabinose, and fucose, according to the peaks in the sample and their peak time, it can be compared and judged that the sample is composed of glucose and arabinose, such as figure 2 As shown, the molar ratio of glucose and arabinose can be calculated to be 4-10:1.

[0025] (2) High performance gel exclusion chromatography to detect its molecular weight

[0026] image 3 In order to use high performance gel exclusion chromatography to determine the molecular weight spectrum of Maca polysaccharide, the molecular weight calibration curve is calculated by GPCPostrunAnalys...

Embodiment 3

[0028] Preliminary Study on Immunomodulatory Activity of Maca Polysaccharide

[0029] (1) The effect of Maca polysaccharide on the proliferation of mouse macrophage Raw264.7

[0030] RAW264.7 cells in the logarithmic phase were digested and counted, and the cell density was adjusted to 3×10 4 / ml, seeded on a 96-well plate, 100 μL / well. Place in a CO2 constant temperature incubator and culture overnight. After the cells adhere to the wall, add different concentrations of Maca polysaccharide (final concentrations are 0, 1.56, 3.125, 6.25, 12.5, 25, 50 μg / ml), and do 4 replicates in each well. , and LPS (1 μg / ml) was added as a positive control, the cell morphology was observed under a microscope after stimulation for 48 hours, and the cell concentration was determined by the MTT method. The result is as Figure 4 shown. It can be seen from the figure that Maca polysaccharide can significantly promote the proliferation of Raw264.7 at 1.56, 3.125, 6.25, and 12.5 μg / ml, but th...

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Abstract

The invention provides a preparation method for maca polysaccharide and preliminary study on immunoregulatory activity thereof. The maca polysaccharide of a relatively high purity is extracted and separated by various means such as boiling water extraction, ethanol precipitation and passing DEAE-52 cellulose resin. Mmonosaccharide is detected to comprise glucose and arabinose with a molar ratio of (4-10): 1 and the molecular weight of the monosaccharide is 10000-30000Da. Meanwhile, the polysaccharide extracted by the method provided by the invention has certain promoting action on activity of a mouse macrophage RAW264.7 and expression of TLR4, TLR2 and an inflammatory factor of the mouse macrophage. The invention has characteristics that the process flow is simple, the operation is easy, the cost is relatively low, and large-scale production is facilitated; the polysaccharide extracted by the method provided by the invention has functions of preventing and resisting cancers, resisting aging, resisting oxidization, resisting autoimmune diseases and the like, thereby providing a theoretical basis for further researching and developing polysaccharide new drugs.

Description

technical field [0001] The invention relates to the technical field of medicine. Specifically, the present invention relates to a Maca polysaccharide with immunoregulatory activity and a preparation method thereof. Background technique [0002] Polysaccharides, also known as polysaccharides, are biological macromolecules formed by the polymerization of ten or more monosaccharides linked by α- or β-glycosidic bonds. It has an extremely important role and is one of the four basic substances that constitute organisms. More than 90% of carbohydrates in nature exist in the form of polysaccharides, which are widely present in the cell membranes of higher animals and plants, algae, fungi, bacteria and other organisms. They have many complex functions such as controlling cell division and differentiation, regulating cell growth and aging. In recent years, it has been found that polysaccharides can inhibit tumors, inhibit bacteria and detoxify because they can enhance the immune fu...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61P37/02
Inventor 乐龙阚红金魏伟坤王莹欧瑜尹鸿萍
Owner CHINA PHARM UNIV
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