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Acupuncture serum exosome as well as preparation method and application thereof

A technology of exosomes and serum, applied in the biological field, can solve problems such as incomplete elucidation and complexity of the mechanism of acupuncture and moxibustion

Inactive Publication Date: 2021-10-22
TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Acupuncture, as a physical stimulus, is perceived locally at acupoints and transferred to the lesion to exert its effect. The process is complex, and the mechanism of acupuncture and moxibustion has not been fully elucidated until now.

Method used

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  • Acupuncture serum exosome as well as preparation method and application thereof
  • Acupuncture serum exosome as well as preparation method and application thereof
  • Acupuncture serum exosome as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Establishment of Rheumatoid Arthritis Model

[0046] (1) Grouping

[0047] Forty-five clean-grade male healthy Wastar rats with a body weight of (220±10) g were selected.

[0048] All rats were purchased from the Experimental Animal Center of the Institute of Hygiene and Environmental Medicine, Academy of Military Medical Sciences of the Chinese People's Liberation Army [permit number: SCXK (Army) 2014-0001].

[0049] All rats were given free access to water and diet during feeding, at a room temperature of 25°C and a humidity of 55%.

[0050] The pelleted mouse feed was provided by the Experimental Animal Center of Tianjin University of Traditional Chinese Medicine, and the drinking water was tap water. They were used for the experiment after adaptive feeding in the same standard environment for 1 week.

[0051] Rats meeting the standard of thermal pain were divided into groups using Microsoft Excel random number method, including: saline group (NS group),...

Embodiment 2

[0064] Example 2: Extraction of serum and exosomes

[0065] In the state of rats after anesthesia, abdominal aortic blood samples were collected from rats with successful modeling (acupuncture 14 times, 27 days after modeling);

[0066] After the blood sample was left to stand for 2 hours, it was centrifuged in a centrifuge at 4°C and 2000rpm for 10 minutes;

[0067] After centrifugation, the supernatant was taken as serum, which was divided into EP tubes, and 500 μL of serum was divided into each EP tube, and all were stored in a -80°C refrigerator for later use;

[0068] S3. Extraction of exosomes

[0069] Take the serum frozen in step S2 and fully thaw it in a refrigerator at 4°C;

[0070] After the serum is thawed, centrifuge in a centrifuge at 4°C and 3000gpm for 15 minutes; after centrifugation, take the supernatant and filter, and collect the filtrate;

[0071] Add a precipitant to the filtrate at a mixing ratio of 4:1 and mix evenly;

[0072] Place the mixed mixtur...

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Abstract

The invention belongs to the technical field of biology, and discloses an acupuncture serum exosome and a preparation method and application thereof. The preparation method comprises the steps that S1, a rheumatoid arthritis model is established, specifically, a complete Freund adjuvant is injected to the sole of the right posterior foot of a rat to cause inflammation, after 24 hours, interventional electro-acupuncture treatment is conducted on the rat, the sole pain threshold of the rat is measured by using a full-automatic thermal radiation stimulator, and whether the rheumatoid arthritis model is successfully molded or not according to a pain threshold measurement result is judged; S2, serum extraction is conducted, specifically, abdominal aorta blood sample collection is conducted on the successfully molded rat, the blood sample is allowed to stand for 2 hours, centrifuging is conducted for 10 minutes in a centrifugal machine with the temperature of 4 DEG C and the speed of 2000rpm, after centrifugation, supernate is taken as serum and subpackaged in EP tubes, 500 microliters of serum is subpackaged in each EP tube, and all the EP tubes are put into a refrigerator at the temperature of -80 DEG C to be cryopreserved for use; and S3, exosome extraction is conducted, specifically, exosome is extracted from the serum extracted in the step S2 to obtain the acupuncture-like serum exosome capable of being used for treating rheumatoid arthritis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an acupuncture serum exosome and its preparation method and application. Background technique [0002] Rheumatoid arthritis is an autoimmune disease of unknown etiology characterized by chronic synovial inflammation of multiple joints throughout the body, which can even lead to joint destruction and deformity. Rheumatoid arthritis has a high incidence rate, high recurrence rate, and high disability rate. Currently, the main therapeutic drugs include non-steroidal anti-inflammatory drugs, corticosteroids, disease-modifying anti-rheumatic drugs and biological agents, etc., but long-term use of these drugs will cause Lead to a variety of side effects and adverse reactions, such as myocardial infarction, peptic ulcer, poor wound healing, liver and kidney insufficiency, etc. [0003] Traditional Chinese medicine has a long-standing understanding of rheumatoid arthritis. Rheum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078A61K35/16A61P29/00A61P19/02
CPCC12N5/0634A61K35/16A61P29/00A61P19/02C12N2509/00C12N2509/10
Inventor 陈波李明月郭义陈泽林李柠岑李牧洋王婷婷
Owner TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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