Use of lactooligosaccharides in preparation of drugs or foods for treating or preventing NEC by alleviating intestinal hypoxia injury
A technology of oligosaccharides and milk sources, which can be used in the field of medicine or food for the treatment or prevention of NEC, milk-derived oligosaccharides, and can solve problems such as incomplete clarity
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Embodiment 1
[0044] Example 1-Establishment of hypoxia injury model of intestinal cells
[0045] Cell line:
[0046] Caco-2, human colorectal adenocarcinoma cells; IEC-6, rat small intestinal crypt epithelial cells, were purchased from the Cell Bank of the Chinese Academy of Sciences. The cells are all DMEM high glucose complete medium at 37℃, containing 5% CO 2 Culture in the cell incubator.
[0047] Cell processing:
[0048] 2×10 4 The concentration of pieces / mL was inoculated into 96-well plates and cultured for 24h. Then replace with 200 μM CoCl 2 Medium, 100μL per well, another control group and a physical hypoxia treatment group (no CoCl added to the medium 2 Solution, add the same volume of PBS), each treatment group set 6 parallel. The control group and the chemical hypoxia treatment group were placed in an incubator, and the physical hypoxia treatment group was treated as described above and placed in an incubator for different periods of time.
[0049] The effect of different hypoxia me...
Embodiment 2
[0069] Example 2-The mitigation effect of milk-derived oligosaccharides on intestinal cell hypoxia damage
[0070] Taking acid milk-derived oligosaccharides as the research object, eight milk-derived oligosaccharides were selected (see Image 6 , Respectively are: neutral oligosaccharide lactoyl-N-tetraose (LNT); carbon chain backbone is lactose, 3'-sialyllactose (3'-SL), 6'-sialic acid residue carrying a single sialic acid residue Acid lactose (6'-SL); carbon chain backbone is LNT or LNnT, sialic acid-lactyl-N-tetraose a (LSTa), sialic acid-lactyl-N-tetraose with a single sialic acid residue b(LSTb), sialic acid-lactoyl-N-tetraose c (LSTc); carbon chain backbone is LNT, disialyl-lactoyl-N-tetraose (DSLNT) carrying two sialic acid residues; and as In contrast, the carbon chain backbone is lactose, which carries a fucosyl 2'-fucose lactose (2'-FL). The aim is to study the difference of milk-derived oligosaccharides with different carbon chain skeletons and the number of sialic ac...
Embodiment 3
[0089] Example 3-The alleviating effect of milk-derived oligosaccharides on necrotizing enterocolitis in mice
[0090] Experimental animals used in this example: 60 SPF seven-day-old C57BL / 6 mice (male and female) were purchased from Beijing Weitong Lihua Experimental Animal Co., Ltd. The formula design of rat milk substitute for feeding mice is shown in Table 1.
[0091] Table 1 Mouse milk substitute formula design
[0092]
[0093] 100mL mouse milk substitute: 10g Nestlé formula milk powder + 7g protein powder + 47.5mL 20% medium and long chain fatty acid milk + 2.6g lactose, dilute to 100mL, after dissolving in warm water bath, store at 4°C.
[0094] Mouse milk replacer containing 4mg / mL 2’-FL: Dissolve 120mg of 2’-FL in 30mL mouse milk replacer, store at 4°C after mixing, and heat with warm water before each use.
[0095] Mouse milk replacer containing 2mg / mL 3'-SL+6'-SL: Add 600μL of 100mg / mL 3’-SL+6’-SL mixed solution to 29.4mL of mouse milk replacer, and store at 4℃ after mixing...
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