Dendrobium-derived exosome-like vesicles as well as preparation and application thereof in improving alopecia
A technology of exosomes and dendrobium, applied in the direction of medical preparations containing active ingredients, pharmaceutical formulas, drug combinations, etc., to achieve good biocompatibility, high promotional effect, and the effect of improving alopecia
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Embodiment 1
[0034] Example 1: Determination of the content of Dendrobium-derived exosome-like vesicles prepared by differential centrifugation combined with density gradient centrifugation
[0035] For the convenience of understanding, the present invention compares the content of exosome-like vesicles from different sources of Dendrobium. Herb Dendrobium, Dendrobium Butterfly, the parts are fresh stems.
[0036] Steps:
[0037] 1) successively pulverize 100g of above-mentioned 9 kinds of dendrobium fresh stems to obtain dendrobium juice;
[0038] 2) The juice was centrifuged at 4°C, 500×g for 20 minutes to obtain a supernatant. Then, the supernatant was centrifuged again at 4° C. and 2,000×g for 30 minutes to obtain a supernatant.
[0039] 3) After successively adding 0.5 ml of 80% sucrose aqueous solution, 1 ml of 30% sucrose aqueous solution and 32 ml of supernatant liquid to remove impurities in a 35 ml ultracentrifuge tube, the mixture was ultracentrifuged at 4° C. and 80,000×g fo...
Embodiment 2
[0045] Example 2: Preparation of Dendrobium-derived exosome-like vesicles
[0046] The invention provides a method for preparing exosome-like vesicles derived from dendrobium as the active ingredient. The preparation raw material is: fresh stems of Dendrobium candidum (DENDROBIUMOFFICINALE).
[0047] Steps:
[0048] 1) Obtain the dendrobium juice by pulverizing the fresh stem of Dendrobium officinale;
[0049] 2) The juice was centrifuged at 4° C., 1000×g for 10 minutes to obtain a supernatant. Then, the supernatant was centrifuged again at 4° C. and 4,000×g for 20 minutes to obtain a supernatant.
[0050] 3) After successively adding 0.5 ml of 80% sucrose aqueous solution, 1 ml of 30% sucrose aqueous solution and 32 ml of supernatant liquid to remove impurities in a 35 ml ultracentrifuge tube, the mixture was ultracentrifuged at 4° C. and 80,000×g for 4 Hour. After ultracentrifugation, the Dendrobium-derived exosome-like vesicles were located between the 80% sucrose layer...
Embodiment 3
[0053] Example 3: Preparation of Dendrobium-derived exosome-like vesicles
[0054] The invention provides a method for preparing exosome-like vesicles derived from dendrobium as an active ingredient. The preparation raw material is: fresh stems of Dendrobium nobile (DENDROBIUM NOBILE).
[0055] Steps:
[0056] 1) Obtain the dendrobium juice by pulverizing the fresh stem of Dendrobium officinale;
[0057] 2) The juice was centrifuged at 4°C, 500×g for 20 minutes to obtain a supernatant. Then, the supernatant was centrifuged again at 4° C. and 2,000×g for 30 minutes to obtain a supernatant.
[0058] 3) After successively adding 0.5 ml of 80% sucrose aqueous solution, 1 ml of 30% sucrose aqueous solution and 32 ml of supernatant liquid to remove impurities in a 35 ml ultracentrifuge tube, the mixture was ultracentrifuged at 4° C. and 80,000×g for 4 Hour. After ultracentrifugation, the Dendrobium-derived exosome-like vesicles were located between the 80% sucrose layer and the ...
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