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Horseradish tree leaf polysaccharide MLP100-3 component with anti-inflammatory activity and purifying technology and application thereof

A technology of Moringa leaves and polysaccharides, which is applied in anti-inflammatory agents, drug combinations, allergic diseases, etc., can solve the problems of strict alkaline extraction conditions, difficulty in achieving expected results, and poor water solubility of polysaccharides, and achieve good anti-inflammatory activity, Good extraction effect, the effect of inhibiting the generation of cell inflammatory factor TNF-α

Inactive Publication Date: 2018-08-17
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, Chen Ruijiao proposed to use 1:20 solid-liquid ratio, 80 ℃ water bath to extract 3 times, each 1.5h, can make the extraction rate of Moringa polysaccharide reach 15.86%, although this method is simple in process, it takes a long time to extract; Relevant studies have proved that hot water extraction-dilute alkali solution can also be used to extract Moringa polysaccharides, but the polysaccharides obtained from the extraction are poor in water solubility, and the extraction conditions of the alkali method are strict, so it is difficult to achieve the expected effect

Method used

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  • Horseradish tree leaf polysaccharide MLP100-3 component with anti-inflammatory activity and purifying technology and application thereof
  • Horseradish tree leaf polysaccharide MLP100-3 component with anti-inflammatory activity and purifying technology and application thereof
  • Horseradish tree leaf polysaccharide MLP100-3 component with anti-inflammatory activity and purifying technology and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of Moringa leaf polysaccharide MLP100-3

[0026] The extraction and purification process of the Moringa leaf polysaccharide MLP100-3 of the present embodiment:

[0027] 1) Weigh 100g of moringa leaves that are pulverized and dried, add 2000 mL of water according to the material-to-liquid ratio of 1:20, and extract for 120 min at 80 ° C, add 3 times the volume of dehydrated alcohol to precipitate overnight, centrifuge, and redissolve after concentrating. . The Sevag method (chloroform:n-butanol=4:1) was used for deproteinization to obtain the crude polysaccharide fraction of Moringa oleifera leaves, which was repeated 4 times until no white flocculent precipitate was precipitated.

[0028] 2) Separation and purification by anion exchange column chromatography

[0029] The Moringa oleifera crude polysaccharide extract obtained in step 1) was separated by DEAE-Sepharose Fast Flow ion exchange column chromatography, and eluted with distilled water, 0...

Embodiment 2

[0033] Example 2 Toxicity test of Moringa polysaccharide MLP100-3 on mouse peritoneal macrophage RAW264.7

[0034] Set the number of cells to 6 x 10 4 The mouse peritoneal macrophages RAW264.7 / well were seeded in a 96-well plate, 100 μL / well. The blank group, the stimulation group, and the sample groups with different concentrations (25 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL) were set up, respectively, in a carbon dioxide incubator (37 °C, 5% CO 2 ) After culturing for 24 h, add 10 μL of 5 mg / mL MTT to each well, incubate in the dark for 4 h, then add 100 μL of DMSO, shake on a shaker for 10 min, and measure the absorbance at 570 nm. The result is as follows figure 2 shown.

[0035] like figure 2 As shown, the Moringa leaf polysaccharide component MLP100-3 has no toxic effect on mouse peritoneal macrophage RAW264.7 in the concentration range of 25-100 μg / mL, and when the concentration of MLP100-3 reaches 200 μg / mL, it has no toxic effect on the cells. toxic side effect. ...

Embodiment 3

[0036] Example 3 Inhibitory effect of Moringa leaf polysaccharide MLP100-3 on the production of inflammatory cells RAW264.7NO

[0037] Set the number of cells to 6 x 10 4 The mouse peritoneal macrophages RAW264.7 / well were inoculated in a 96-well plate, 100μL / well. The blank group, the stimulation group, and the sample groups with different concentrations (25 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL) were set up, respectively, in a carbon dioxide incubator (37 °C, 5% CO 2 ) After culturing for 2 hours, lipopolysaccharide (LPS) with a final concentration of 1 μg / mL was added, and the culture was continued for 24 hours. The NO content in the supernatant of the culture medium was measured using the NO kit, 50 μL culture medium + 50 μL Griess Reagent I + 50 μL Griess Reagent II, and the absorbance was measured at 540 nm immediately. Using different concentrations of sodium nitrite as the standard curve, the NO concentration produced in each group was calculated. The result is as f...

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Abstract

The invention relates to a horseradish tree leaf polysaccharide MLP100-3 component with anti-inflammatory activity. The monosaccharide of the horseradish tree leaf polysaccharide MLP100-3 is preparedfrom rhamnose, arabinose, xylose, mannose, glucose and galactose according to a mass ratio proportion of (1.2 to 1.3) to (4.5 to 5.4) to (4.2 to 5.5) to (3.4 to 4.0) to (5.6 to 6.6) to (5.9 to 6.5). The invention further provides a purifying method of the horseradish tree leaf polysaccharide MLP100-3. The horseradish tree leaf polysaccharide MLP100-3 disclosed by the invention show better capacityof inhibiting NO and inflammatory factor TNF-alpha generation in mouse peritoneal macrophage RAW264.7 in-vitro inflammation model. The horseradish tree leaf polysaccharide MLP100-3 disclosed by the invention can be applied to adjusting of body immune function and development and utilization of anti-inflammatory medicine.

Description

technical field [0001] The invention belongs to the field of natural product chemistry, and in particular relates to a Moringa leaf polysaccharide MLP100-3 extracted and purified from natural Moringa oleifera and its use for anti-inflammatory and immune regulation. Background technique [0002] Moringa, also known as drumstick tree and horseradish tree, is native to northern India. It is a perennial tropical and subtropical deciduous tree. There are 14 varieties worldwide and are widely planted in tropical and subtropical regions of Asia, Africa and Africa. . The leaves, flowers, seeds, roots, and seeds of Moringa oleifera can be eaten directly and are high in calcium, protein, fiber, and lipids. In addition to its high edible value, Moringa can also enhance human immunity, promote physical health, and prevent diseases. It is used to treat diseases in India, Africa and other countries or regions, and is known as a health care treasure. It has been clinically proven that Mo...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61P29/00A61P37/02
CPCA61P29/00A61P37/02C08B37/0003C08B37/006
Inventor 王磊崔璨陈晓阳
Owner SOUTH CHINA AGRI UNIV
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