Catharanthus roseus-sourced exosome nano-vesicle and application thereof in immunoregulation

A technology of nanovesicles and periwinkle, which is applied in the field of medicine to achieve the effect of strong antimicrobial and antitumor activity and strong clinical application value

Pending Publication Date: 2022-05-27
于荣敏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the technical problems currently faced in this field, the present invention provides a vinca exosome nanovesicle with immune activity;

Method used

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  • Catharanthus roseus-sourced exosome nano-vesicle and application thereof in immunoregulation
  • Catharanthus roseus-sourced exosome nano-vesicle and application thereof in immunoregulation
  • Catharanthus roseus-sourced exosome nano-vesicle and application thereof in immunoregulation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Extraction and isolation of exosome-like nanovesicles derived from periwinkle leaves

[0055] Enzymatic hydrolysis of periwinkle leaves: Pick fresh periwinkle leaves to ensure that the outline of the leaves is complete, green and flat, without curling and yellowing, and no traces of insect bites. The leaves were rinsed 3 times with tap water and 3 times with distilled water, dried with filter paper, and weighed for recording. Prepare an enzymatic hydrolysis solution containing cellulase (3 g per 100 ml) and pectinase (0.2 g per 100 ml), pH between 5.75 and 5.80. Add 100 ml of enzymatic hydrolysis solution per 2 g of leaves and incubate at 25°C for at least 12 h in the dark at 100 rpm on a shaker. Let stand for 30 minutes, and the supernatant liquid was collected and centrifuged at 1000 rpm for 20 minutes to remove impurities. Then centrifuge at 10,000 rpm for 40 min at 4°C, repeat the centrifugation 3 times, take the supernatant, and concentrate it to 100 ml...

Embodiment 2

[0060] Example 2 Extraction and isolation of exosome-like nanovesicles derived from periwinkle cambium stem cells

[0061] Cell pre-culture and preparation before loading: Take well-grown periwinkle stem cells, add 2 g of cells per 100 ml of liquid medium, and culture in suspension at 120 rpm for 7 days in complete darkness. The liquid medium contained 20 g / L sucrose, 2.0 mg / L α-naphthaleneacetic acid (NAA) and 4.43 g / L MS medium, and the pH was between 5.75 and 5.80. After 7 days of suspension culture, the plant cells were filtered with gauze and the medium was collected. 10000×g, 4°C, 10min, centrifuge 3 times to remove impurities, and collect the supernatant. The filtrate was concentrated to 100ml with a hollow fiber dialysis tube.

[0062] One-Step Sucrose Pad Ultracentrifugation: Add 7 ml of 0.971M sucrose to the bottom of the ultracentrifuge tube, then add the sample to fill the centrifuge tube. Centrifuge at 150,000 × g for 90 min at 4°C. The sucrose layer was colle...

Embodiment 3

[0066] Example 3 Extraction and isolation of exosome-like nanovesicles derived from periwinkle callus

[0067] Cell pre-culture and preparation before loading: Take well-grown periwinkle mosaic callus, add 4 g of cells per 100 ml of liquid medium, and culture in suspension at 120 rpm for 7 days in complete darkness. The liquid medium contained 20 g / L sucrose, 2.0 mg / L α-naphthaleneacetic acid (NAA) and 4.43 g / L MS medium, and the pH was between 5.75 and 5.80. After 7 days of suspension culture, the plant cells were filtered with gauze and the medium was collected. 10000×g, 4°C, 10min, centrifuge 3 times to remove impurities, and collect the supernatant. The filtrate was concentrated to 100ml with a hollow fiber dialysis tube.

[0068] One-Step Sucrose Pad Ultracentrifugation: Add 7 ml of 0.971M sucrose to the bottom of the ultracentrifuge tube, then add the sample to fill the centrifuge tube. Centrifuge at 150,000 × g for 90 min at 4°C. The sucrose layer was collected, 20 ...

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Abstract

The invention relates to a catharanthus roseus exosome nano-vesicle with immunocompetence and a preparation method of the catharanthus roseus exosome nano-vesicle. The Catharanthus roseus exosome nano-vesicles comprise Catharanthus roseus leaf source exosome-like nano-vesicles, Catharanthus roseus stem cambium stem cell source exosome-like nano-vesicles, Catharanthus roseus leaf callus source exosome-like nano-vesicles and Catharanthus roseus stem source exosome-like nano-vesicles. The extracted Catharanthus roseus plant-derived exosome vesicles can be maintained stable in an artificial simulated digestive system, and still exert immunoregulatory activity under strong acid, strong alkali and enzyme digestion regulation.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a periwinkle plant-derived nanovesicle with immunomodulatory activity. Background technique [0002] Periwinkle is a perennial herb of the oleander family. It is widely distributed in the south of the Yangtze River. It is a common street plant with a long flowering period and can be planted all year round. The indole terpenoid alkaloids contained in vinca have been found to have excellent antitumor activity. Currently, vinblastine, vincristine, vindesine and vinorelbine have been marketed and used for clinical tumor treatment. However, vinca alkaloids are highly toxic, and their injections can cause peripheral phlebitis after intravenous administration, as well as adverse reactions such as nausea and vomiting, neurotoxicity, and bone marrow suppression. The toxic and side effects of vinca alkaloids greatly limit their clinical application. [0003] Exosomes are nanovesicles re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04A61K36/24A61K9/00A61P37/02A61P35/00A61K135/00
CPCC12N5/04A61K36/24A61K9/0053A61P37/02A61P35/00A61K2236/19A61K2236/51A61K2236/53
Inventor 于荣敏朱建华欧筱争
Owner 于荣敏
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