CN-118320022-B - A nanometer vesicle containing quercetin and capable of protecting alcoholic liver injury, and its preparation method and application

Abstract

The invention discloses a anoectochilus roxburghii-derived nano vesicle containing quercetin and capable of protecting alcoholic liver injury, and preparation and application thereof. Quercetin is an active ingredient with antioxidant and anti-inflammatory effects contained in anoectochilus roxburghii, however, the in-vivo curative effect is reduced due to lower water solubility and bioavailability, so that the clinical curative effect is limited. The quercetin obtained by the method for separating the anoectochilus roxburghii-derived nano vesicles has the advantages of simplicity and rapidness in operation, and the lipid bilayer of the exosome-like vesicles is used as a natural protection film of the quercetin, so that the stability of the quercetin can be maintained for a long time. When the anoectochilus roxburghii-derived nanocapsule is used for HepG2 cells, the liver index of a mouse can be reasonably regulated and controlled, adverse reactions such as hepatocyte edema, steatosis, lipid drop accumulation and the like caused by taking of machine type alcohol can be effectively prevented or inhibited, and the anoectochilus roxburghii-derived nanocapsule has remarkable curative effects on alcoholic liver diseases.

Inventors

• ZHANG XI
• Zou Xiu
• CHEN JINGHUA
• GAO DANDAN
• LU SHI
• CHEN GUANYU
• XU LILAN

Assignees

• 福建医科大学

Dates

Publication Date

20260508

Application Date

20240227

Claims (1)

1. 1. The application of the anoectochilus formosanus source nano-vesicles in preparing the medicine for preventing and treating the alcoholic liver disease is characterized in that the preparation method of the anoectochilus formosanus source nano-vesicles comprises the following steps: s1, squeezing fresh whole herb of anoectochilus roxburghii by a juicer, and filtering by gauze to obtain filtrate; s2, carrying out high-speed centrifugation on the filtrate obtained in the step S1, and discarding the precipitate to obtain supernatant; S3, performing ultracentrifugation on the supernatant obtained in the step S2, and collecting precipitate, and re-suspending with PBS to obtain anoectochilus formosanus-derived nano vesicle suspension containing quercetin and capable of protecting alcoholic liver injury; wherein, the juice squeezing process needs to mix precooled PBS or PBS ice cubes with the fresh whole herb of anoectochilus roxburghii; Wherein the alcoholic liver disease is alcoholic liver injury and alcoholic fatty liver.

Description

A nanometer vesicle containing quercetin and capable of protecting alcoholic liver injury, and its preparation method and application Technical Field The invention relates to the technical field of separation and extraction of plant exosome-like vesicles and research of anticancer drugs, in particular to extraction of anoectochilus formosanus-derived nano vesicles containing active pharmaceutical ingredients and feasibility research of using the anoectochilus formosanus-derived nano vesicles as liver protecting drugs. Background The exosome is a nanoscale vesicle actively secreted by cells, has the functions of enhancing permeation retention (enhanced permeability and retention, EPR) effect and protecting contents, also carries relevant genetic information (such as protein, nucleic acid and the like) of parent cells, has good targeting ability on homologous cells, can participate in intercellular communication, can even cross blood brain barrier and bypass a cell P-glycoprotein drug efflux system, and has certain advantages in research on administration in brain and reduction of cell drug resistance. In addition, the exosome contains lipid bilayer, so that the exosome can effectively protect the internal active ingredients, ensure that the exosome is not decomposed before reaching a target position, and can be used as a drug carrier for drug delivery. In recent years, as researchers further understand and explore exosomes, they find great potential in drug delivery and disease treatment. However, currently, exosomes as drug carriers are mainly derived from human cells and human body fluids, and these sources have low exosomes content, high separation cost and complicated purification methods, and tumor-derived exosomes may trigger tumor diffusion or immune response of the organism, thereby limiting their application in drug delivery studies. Plant-derived nanovesicles (Plant-derived nanovesicles, PDNVs) are nanoscale vesicles secreted by Plant cells, are remarkably similar to exosomes in morphology and function, and can be used as emerging nanoparticles to provide a new idea for solving the problems. PDNVs has simple separation and extraction process and low cost, carries various lipid, protein, nucleic acid and parent bioactive molecules, can play a synergistic treatment role when acting on different passages, has better curative effect on disease treatment compared with single active ingredients extracted from plants, has better stability in gastrointestinal tract when being taken as exosome-like vesicles with lipid bilayer structures, has phospholipid structures capable of becoming natural protective barriers to prevent degradation or inactivation of internal active ingredients and improve bioavailability, has low immunogenicity, high safety and natural ingredients, has good biosafety in vivo, can be taken as a novel nano administration dosage form to realize rapid and effective treatment on the disease, has good oral bioavailability, and has stability in gastrointestinal tract and is not easy to be digested and degraded by gastric juice, pancreatic juice and bile liquid to lose activity. Based on the advantages, the research application of the plant exosome-like vesicles in a drug carrier has great potential. Anoectochilus roxburghii (Anoectochilus roxburghii) is perennial herb of Anoectochilus of Orchidaceae, and is a precious plant in Chinese medicinal materials, and contains active component quercetin with antioxidant and antiinflammatory effects. Research shows that anoectochilus roxburghii has therapeutic effects on liver injury, nonalcoholic fatty liver disease, autoimmune hepatitis, chronic hepatitis B and the like caused by carbon tetrachloride (CCl 4). However, anoectochilus formosanus has problems in oral administration, such as high cost, easy inactivation of active ingredients during processing, poor bioavailability, and the like. Quercetin is an active ingredient with antioxidant and antiinflammatory effects contained in anoectochilus formosanus, and can restore intracellular glutathione level, reduce oxidative stress of cells, and reduce formation of lipid peroxide malondialdehyde. However, its lower water solubility and bioavailability reduce its in vivo therapeutic effect, making its clinical therapeutic effect limited. The problems greatly limit the drug-forming property and clinical application of the anoectochilus formosanus in preparing oral dosage forms. The anoectochilus roxburghii-derived nano vesicle has a lipid bilayer structure similar to an exosome, is a natural protective film, and can reduce the influence of external environment on the internal components of the anoectochilus roxburghii-derived nano vesicle. In addition, the presence of the lipid bilayer also increases the stability of quercetin to some extent. Disclosure of Invention The invention aims to provide a anoectochilus roxburghii-derived nano vesicle containing quercetin and capable of protecting alcoholic liver i