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CN-122011231-A - Preparation method and application of Morchella esculenta polysaccharide

CN122011231ACN 122011231 ACN122011231 ACN 122011231ACN-122011231-A

Abstract

The invention relates to the field of application of fungus polysaccharide, and discloses a preparation method and application of Morchella polysaccharide. The preparation method comprises the following steps of 1) leaching dried Morchella esculenta powder by water to obtain leaching solution, precipitating the leaching solution by alcohol to obtain precipitate, 2) adding water to the precipitate for re-dissolution, removing proteins, dialyzing and freeze-drying to obtain Morchella esculenta crude polysaccharide extract, and 3) dissolving the Morchella esculenta crude polysaccharide extract in water, and purifying by adsorption chromatography to obtain Morchella esculenta polysaccharide. The morchella polysaccharide has the protection effect on alcoholic gastric injury in vivo and in vitro, and the gastric tissue inflammatory reaction is regulated by regulating the NF-kb path, so that the reference value is provided for the development and application of the morchella polysaccharide.

Inventors

  • CHEN HANGJUN
  • ZHANG JIANMING
  • GAO HAIYAN
  • LIU RUILING
  • WU WEIJIE
  • CHEN HUIZHI
  • FANG XIANGJUN
  • YIN MING
  • WANG GUANNAN
  • HAN YANCHAO

Assignees

  • 浙江省农业科学院

Dates

Publication Date
20260512
Application Date
20260331

Claims (10)

  1. 1. The preparation method of Morchella polysaccharide is characterized by comprising the following steps: Step 1) leaching dried Morchella powder by water to obtain leaching solution; Precipitating the leaching solution with alcohol to obtain a precipitate; step 2) adding water to the precipitate for re-dissolution, removing protein, and then dialyzing and freeze-drying to obtain Morchella crude polysaccharide extract; And 3) dissolving the Morchella crude polysaccharide extract in water, and purifying by adsorption chromatography to obtain Morchella polysaccharide.
  2. 2. The preparation method of claim 1, wherein the step 1) is characterized in that the dried morchella powder is subjected to ultrasonic leaching by water, the solid-to-liquid ratio of the ultrasonic leaching is 1:10-50 g/ml, the leaching temperature is 30-70 ℃, the leaching time is 20-60min, and the power is 100-400W.
  3. 3. The process of claim 1 wherein in step 1) the leaching solution is concentrated by rotary evaporation and then subjected to alcohol precipitation with three volumes of ethanol; the alcohol precipitation temperature is 1-8 ℃, and the alcohol precipitation time is 10-16h; And centrifuging after the alcohol precipitation is finished, wherein the centrifuging condition is 3000-5000r/min, and the centrifuging time is 5-15min.
  4. 4. The method according to claim 1, wherein the dialysis in step 2) is performed using a dialysis bag of 3000-8000Da for a dialysis time of 48-72 h.
  5. 5. The preparation method of the Morchella esculenta polysaccharide according to claim 1, wherein the Morchella esculenta polysaccharide extract obtained in the step 3) is dissolved in water, and then passes through a 0.45 μm microporous filter membrane, and then passes through an ion exchange column, and then adopts sodium chloride solution for gradient elution, eluent is collected, and after dialysis and freeze-drying, primarily purified polysaccharide is obtained, and the primarily purified Morchella esculenta polysaccharide is dissolved in water, and then passes through a gel column, and then adopts distilled water for elution, eluent is collected, and freeze-dried, so that the Morchella esculenta purified polysaccharide is obtained.
  6. 6. The preparation method according to claim 5, wherein the solution used in the step 2) is a solution containing 0-0.5 mol/L sodium chloride, the elution speed is 3-5 ml/min, and the elution speed of distilled water is 0.9-1.1 ml/min.
  7. 7. The Morchella polysaccharide prepared by the preparation method of any one of the claims 1-6, which is characterized in that the Morchella polysaccharide has a weight average molecular weight of 24.759 kDa, a number average molecular weight of 38.634kDa, a polydispersity Mw/Mn of 1.560, and a molar ratio of glucose to mannose to galactose of 61.61:26.37:12.02.
  8. 8. The use of Morchella polysaccharide according to claim 7, characterized in that the Morchella polysaccharide is used for preparing a medicament for preventing and protecting gastric mucosal injury.
  9. 9. The use of claim 8, wherein the gastric mucosal lesion comprises ethanol-induced gastric mucosal hemorrhage, edema, erosion.
  10. 10. The use of claim 8, wherein the prevention of gastric mucosal lesion is achieved by alleviating inflammatory symptoms and modulating oxidative stress.

Description

Preparation method and application of Morchella esculenta polysaccharide The invention field: the invention belongs to the field of polysaccharide preparation and application, and particularly relates to a preparation method and application of Morchella polysaccharide. The background technology is as follows: gastric ulcers are a common digestive system disease in the world, severely affecting people's quality of daily life. Among them, helicobacter Pylori (HP) infection, drinking, smoking, nonsteroidal anti-inflammatory drugs and other external factors may cause gastric ulcer formation. Drinking, a common factor, often affects multiple parts of the gastrointestinal tract, directly or indirectly causing gastric lesions (e.g., oedema, erosion, bleeding, and necrosis). In clinical treatment, the treatment of alcoholic gastric ulcer mainly comprises proton pump inhibitor, gastric acid neutralizer, digestive enzyme preparation and the like, however, long-term use of the medicines usually involves a plurality of complications such as acute kidney injury, gastric mucosa result and function change, drug resistance and the like. Accordingly, natural product alternatives are of great interest for the treatment of gastric mucosal lesions. Morchella is taxonomically belonging to Morchella dill.ex Pers of Morchellaceae (Morchell-aceae) of Ascomycotina (Ascomycotina). Modern pharmacological research shows that Morchella has immunoregulation and physiological activity. The medicinal value of Morchella is recorded in the compendium of Ben Cao at the earliest, and the Morchella has the effects of improving intestines and stomach, resolving phlegm and regulating qi. The medicinal value of Morchella esculenta in the diet therapy herbal is described as sweet and cold without toxicity, phlegm reduction, qi regulation and stomach benefiting, and at present, although researches have been made to disclose the activities of the Morchella esculenta polysaccharide such as immunoregulation, anti-tumor and the like, the protection effect and specific mechanism of polysaccharide components with specific structural characteristics on alcoholic gastric mucosal injury have not been reported yet. The invention firstly separates and purifies Morchella to obtain the uniform polysaccharide MEP-1 with specific molecular weight and monosaccharide composition, has dual effects of anti-inflammatory and antioxidant, prevents and treats alcoholic gastric injury through NF- κB passage, and provides a new candidate medicament for gastric mucosa protection. Disclosure of Invention In order to solve the problem of extracting polysaccharide for protecting gastric mucosa from Morchella esculenta. The invention provides a preparation method and application of Morchella polysaccharide. A preparation method of Morchella polysaccharide comprises the following steps: Step 1) leaching dried Morchella powder by water to obtain leaching solution; Precipitating the leaching solution with alcohol to obtain a precipitate; step 2) adding water to the precipitate for re-dissolution, removing protein, and then dialyzing and freeze-drying to obtain Morchella crude polysaccharide extract; Step 3) dissolving the Morchella crude polysaccharide extract in water, and purifying by adsorption chromatography to obtain Morchella polysaccharide; optionally, the step 1) is to carry out ultrasonic leaching on the dried Morchella powder by water, wherein the solid-liquid ratio of the ultrasonic leaching is 1:10-50 g/ml, the leaching temperature is 30-70 ℃, the leaching time is 20-60min, and the leaching power is 100-400W. Optionally, in the step 1), the leaching solution is concentrated by rotary evaporation, and then three times of volume of ethanol is adopted for alcohol precipitation; the alcohol precipitation temperature is 1-8 ℃, and the alcohol precipitation time is 10-16h; And centrifuging after the alcohol precipitation is finished, wherein the centrifuging condition is 3000-5000r/min, and the centrifuging time is 5-15min. Optionally, in the step 2), a dialysis bag with a speed of 3000-8000Da is used for dialysis, and the dialysis time is 48-72 h. 5. The preparation method of the Morchella esculenta polysaccharide according to claim 1, wherein the Morchella esculenta polysaccharide extract obtained in the step 3) is dissolved in water, and then passes through a 0.45 μm microporous filter membrane, and then passes through an ion exchange column, and then adopts sodium chloride solution for gradient elution, eluent is collected, and after dialysis and freeze-drying, primarily purified polysaccharide is obtained, and the primarily purified Morchella esculenta polysaccharide is dissolved in water, and then passes through a gel column, and then adopts distilled water for elution, eluent is collected, and freeze-dried, so that the Morchella esculenta purified polysaccharide is obtained. Optionally, the eluent used in the step 2) is solution containing 0-0.5 mol/L sodium chloride,