Bitter gourd exosome, and extraction method and application of same
An extraction method and exosome technology, applied in the field of medicine, to achieve the effects of high impurity removal rate, blood-brain barrier protection, and reduced penetration
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Embodiment 1
[0029] Example 1: Extraction method of bitter melon exosomes
[0030] See attached figure 1 For the extraction process shown, take 500g of fresh Yunnan mountain wild bitter gourd, remove the seeds and wash three times, each time for 1-3min, and dry after washing;
[0031] After drying, squeeze bitter gourd juice with a fresh juicer, about 200ml, and centrifuge the bitter gourd juice continuously at 4°C, 1000g, 10min; 3000g, 20min; 10000g, 40min; after centrifugation, take the supernatant and discard the precipitate;
[0032] Ultracentrifuge the obtained supernatant, centrifuge at 4°C, 150,000g for 90min in an ultra-fast refrigerated centrifuge, discard the supernatant after centrifugation, take the precipitate, and suspend the precipitate in 1-2ml of PBS buffer;
[0033] The suspension was filtered with a 0.22um filter membrane, and the filtrate was centrifuged again in an ultracentrifuge at 4°C and 150,000g for 90min. After centrifugation, the precipitate was collected, the ...
Embodiment 2
[0037] Example 2: Brain protection of bitter gourd exosomes
[0038] 1. Experimental animals
[0039] Male Sprague-Dawley (SD) rats, weighing 250±20g, clean grade, provided by the Experimental Animal Center of Xuzhou Medical University, experimental animal production license: SCXK (Shanghai) 2018-0006.
[0040] 2. Main reagents
[0041] Bitter melon exosomes: extracted according to the method described in the examples; TTC (2,3,5-triphenyltetrazolium chloride; Shanghai Sangong), Evans blue (EB; Beijing Yinuokai Technology Co., Ltd.), Trichloroacetic acid (Beijing Yinuokai Technology Co., Ltd.), CD54 / ICAM-1 Antibody#4915 (CST), TSG101 Antibody (proteintech), CD63 Antibody (proteintech)
[0042] 3. Instrument
[0043] Ultra-speed refrigerated centrifuge (Beckman Optima L-100XP), transmission electron microscope (Tecnai G2Spirit Twin*Tecnai G2 Spirit Twin), Zeta potential detector (Nicomp380ZLS), small animal anesthesia machine (ABS-100), Tanon electrophoresis instrument (EPS ...
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