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CN-121987607-A - Preparation method and application of ginseng-derived nano-sample particle loaded drug

CN121987607ACN 121987607 ACN121987607 ACN 121987607ACN-121987607-A

Abstract

The invention relates to the technical field of biological medicine and nano preparation, in particular to a preparation method and application of a ginseng-derived nano sample particle loaded medicine. The method comprises the steps of S1, extracting ginseng exosomes, S2, purifying the ginseng exosomes, and S3, preparing drug-loaded nano-particles. The invention uses GENs as oral carrier to solve the problem of low bioavailability of free medicine, and the natural phospholipid bilayer structure of the invention enhances the stability of medicine, which is beneficial to intestinal absorption and liver targeting accumulation. GENs has the double functions of active ingredients and loaded medicines, is obviously stronger than that of single medicines, and synchronously realizes antioxidation, anti-inflammation, anti-fibrosis and anti-apoptosis. The intestinal barrier is rebuilt through repairing ZO-1/Occludin, the enterogenic toxin entering the liver is reduced, the liver injury and the disappearance of fibrosis are improved, a new strategy is provided for the high-valued conversion of ginseng resources, and the safety of plant exosomes is far higher than that of a synthetic carrier.

Inventors

  • YANG CHUN
  • ZHANG YAN

Assignees

  • 北华大学

Dates

Publication Date
20260508
Application Date
20260108

Claims (9)

  1. 1. The preparation method of the ginseng-derived nano-sample particle loaded medicine is characterized by comprising the following steps of: S1, ginseng exosome extraction, namely cleaning ginseng, cutting the ginseng into small pieces, adding PBS buffer solution, homogenizing the ginseng by using a homogenizer, filtering by using gauze, centrifuging to obtain supernatant, centrifuging by using an ultracentrifuge, taking precipitate, re-suspending the precipitate by using PBS, and fully swirling to obtain crude extracted ginseng exosome; s2, purifying the ginseng exosomes, namely placing the crude ginseng exosomes in a sucrose density gradient solution, centrifuging, absorbing a sample layer, supplementing PBS buffer solution, centrifuging, washing, and re-suspending the precipitate by using PBS to obtain pure ginseng exosomes; s3, preparing the drug-loaded nano-particles, namely mixing the purified ginseng exosome solution with the free drug according to the mass ratio, and carrying out intermittent ultrasonic treatment and shaking table incubation overnight to obtain the nano-sample particle-loaded drug.
  2. 2. The method for preparing a drug loaded with ginseng-derived nano-particles according to claim 1, wherein the free drug is ferulic acid.
  3. 3. The method for preparing a ginseng-derived nano-particle-supported drug according to claim 1, wherein the ginseng exosome solution and the free drug are mixed according to a mass ratio of 1:2-4.
  4. 4. The method for preparing a drug loaded with nano-particles derived from ginseng according to any one of claim 1 or 2, wherein the particle size of the drug loaded with nano-particles is 150-200nm, the surface potential is-25 to-30 mV, and the drug loaded with nano-particles has a double-layer membrane structure.
  5. 5. A method of treating liver injury, comprising administering to the liver an agent comprising the subject of claim 4, wherein the agent is administered in combination with the agent, and wherein the agent is administered in combination with the agent.
  6. 6. The use of the nano-particle loaded medicine according to claim 5 for preparing a medicine for repairing liver injury, wherein the nano-particle loaded medicine repairs intestinal barriers by regulating intestinal flora, thereby improving liver microenvironment and realizing liver and intestine bidirectional regulation.
  7. 7. The use of a nanoparticle-loaded drug according to claim 4 for inhibiting expression of fibrotic markers Vimentin and alpha-SMA in hepatic stellate cells, and for reducing collagen deposition, and reversing CCl 4-induced pathological characteristics of hepatic fibrosis in mice.
  8. 8. The use of a nanoparticle-loaded drug according to claim 7 for the preparation of an anti-liver fibrosis drug, wherein the nanoparticle-loaded drug restores liver function by decreasing serum AST/ALT levels.
  9. 9. The method of preparing a drug for repairing intestinal barrier according to claim 4, wherein the drug is used for repairing intestinal villus structural integrity and up-regulating expression of intestinal zonulin ZO-1 and Occludin.

Description

Preparation method and application of ginseng-derived nano-sample particle loaded drug Technical Field The invention relates to the technical field of biological medicine and nano preparation, in particular to a preparation method and application of a ginseng-derived nano sample particle loaded medicine. Background Liver fibrosis is a key pathological process by which chronic liver disease progresses to cirrhosis and liver cancer, and liver-related mortality increases exponentially with increasing fibrosis. It is characterized by increased synthesis of hepatic stellate cell activated extracellular matrix (ECM), leading to excessive deposition of ECM, advanced progression to cirrhosis and even liver cancer, and death induced by hepatic failure in end-stage patients. The early stage clinical symptoms of liver fibrosis are not obvious, are usually middle and late stages when diagnosis is usually confirmed, have the clinical characteristics of large harm, difficult reversion and the like, have the incidence rate rising year by year on the whole world, and are particularly obvious in high-risk groups such as chronic viral hepatitis (e.g. hepatitis B and hepatitis C), nonalcoholic fatty liver disease (MAFLD), alcoholic liver disease and the like. Liver fibrosis is mainly due to the increased deposition of extracellular matrix secreted by hepatic stellate cells, resulting in stiffening of liver texture further developing cirrhosis or even liver cancer. The pathological process of liver fibrosis is complex, and the liver fibrosis has single activation effect of inhibiting hepatic stellate cells, and the oral free medicine has low availability and loss, and the single medicine has limited curative effect. Disclosure of Invention The invention aims to provide a ginseng-derived nano-sample particle loaded medicament, a preparation method and application thereof, wherein the ginseng exosome loaded medicament is used for coordinating the regulation and control of intestinal flora by utilizing the circulatory action of intestinal liver axes, and the liver fibrosis is relieved, so that the problems in the background technology are solved. In order to achieve the above object, in one aspect, the present invention provides a method for preparing a medicament loaded with nano-sample particles derived from ginseng, comprising the steps of: s1, extracting Ginseng Exosomes (GENs): Washing ginseng, cutting into small pieces, adding PBS buffer solution, homogenizing by using a homogenizer, filtering by using gauze, sequentially centrifuging for 30min according to 3000g, centrifuging for 1h according to 10000g, taking supernatant, centrifuging for 1h according to 100000g by using an ultracentrifuge, taking precipitate, re-suspending the precipitate by using PBS, and fully swirling to obtain crude extracted Ginseng Exosomes (GENs); S2, purifying Ginseng Exosomes (GENs): preparing 1mol/L and 2mol/L sucrose solutions, placing a resuspended sample in a super-separation tube, sequentially penetrating through a sample layer, respectively placing the 1mol/L sucrose solution and the 2mol/L sucrose solution, centrifuging for 2 hours at 150000g, absorbing the sample layer, adding PBS (phosphate buffer solution) into the whole tube, centrifuging for 1 hour at 100000g, washing off excessive sucrose, resuspending and precipitating with PBS to obtain a purified ginseng exosome, measuring the protein concentration in the ginseng exosome by using a BCA kit, and storing in a refrigerator at-80 ℃; s3, preparation of drug-carrying exosomes: Mixing Ginseng radix exosome (GENs) solution with free drug (preferably ferulic acid FA) at mass ratio of 1:2-4, intermittently ultrasonic for 1 hr, and shaking overnight at 4 ° to obtain Ginseng radix-derived nano-sample particle-loaded drug (Exo@FA) (particle size of nano-sample particle-loaded drug is 150-200nm, and surface potential is-25-30 mV). According to the invention, ginseng Exosomes (GENs) are taken as natural carriers, and are subjected to sucrose density gradient centrifugation and purification to load drugs (such as ferulic acid FA), so that Exo@FA nano-particles with the particle size of about 191nm are formed. After oral administration, GENs improves the bioavailability of the drug and targets delivery to the liver, and the drug contains active ingredients such as ginsenoside and the like, and plays an anti-inflammatory and anti-oxidation role in cooperation with the loaded drug: liver protection, namely reducing the ROS level in liver cells (H2O 2 model: fluorescence intensity is reduced from 92.1% to 23.7%, LPS model: 16.7% to 4.90%), inhibiting apoptosis (apoptosis rate is reduced from 73.7% to less than or equal to 5%), and improving activity (CCK-8 proliferation rate is obviously increased); anti-fibrosis, inhibiting expression of hepatic stellate cell fibrosis marker Vimentin/alpha-SMA (green fluorescence disappeared), reducing collagen deposition (Sirius red/Masson staining reversal). Next