Model establishment method and application of MRJPs for regulating mouse immunity and intestinal microorganisms
A technology for establishment of intestinal microorganisms and models, applied in the field of model establishment of MRJPs on mouse immunity and intestinal microbial regulation, can solve problems such as lack of understanding, achieve enhanced immunity, enhance immune function, and promote the proliferation of intestinal probiotics Effect
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Embodiment 1
[0052] Example 1: Quality analysis of MRJPs freeze-dried powder
[0053] MRJPs are prepared from fresh royal jelly, separated by ultrafiltration separation equipment and process, freeze-dried, and stored frozen at -20°C. The qualitative analysis of MRJPs by SDS-PAGE electrophoresis showed that there were more than 4 bands including MRJP1, MRJP2, MRJP3 and MRJP5, with molecular weights ranging from 25 to 87 kDa ( figure 1 ).
[0054]The nitrogen determination method was used to quantitatively detect the total protein of MRJPs freeze-dried powder, and the average total protein content was 75.85%; the spectrophotometric method was used to detect the total sugar content of 9.35%; the liquid chromatography method was used to quantitatively detect the 10-HAD content of 0.86%. MRJPs are glycoproteins with N-linked glycosylation sites, which are composed of proteins and sugar chains. Therefore, it is speculated that the glycoprotein content of MRJPs freeze-dried powder is more than...
Embodiment 2
[0055] Example 2: Evaluation of MRJPs for enhancing the immune function of mice
[0056] Female C57BL / 6 mice (6-8 weeks old) were taken, and all mice were housed in an environment with a 12h:12h dark:light cycle. Mice were randomly divided into 4 groups (n=6 / group): (1) control group (CK): MRJPs 0 mg / kg by gavage; (2) M125 group: MRJPs 125 mg / kg by gavage Stomach; (3) M250 group: mice were given MRJPs 250 mg / kg by gavage; (4) M 500: mice were given MRJPs 500 mg / kg by gavage. The control group was given the corresponding dose of normal saline every day, and the mice in the (2)-(4) groups were given the corresponding dose of MRJPs every day, and were weighed once a week until the end of the experiment. All experimental procedures were reported to the ethics committee for approval in accordance with ethical requirements.
[0057] (1) The effect of MRJPs on changes in body weight and organ weight: The mice were weighed after 4 weeks of rearing, and no difference was found in the...
Embodiment 3
[0064] Example 3: Evaluation of the function of MRJPs to promote the proliferation of intestinal probiotics in mice
[0065] Since MRJPs at 500 mg / kg body weight elicited the most potent peripheral and local immune responses in mice, this dose was used later for gut microflora analysis.
[0066] After the mice were fed for 4 weeks, fresh fecal samples were taken from the mice and stored at -80°C until DNA extraction. Total genomic DNA was extracted from mouse fecal samples using the Ezna® Soil DNA Kit and Procedure Extraction. The V3-V4 regions of the 16SrRNA gene were amplified with forward primer (5'-ACTCCTACGGGAGGCAGCA-3') and reverse primer (5'-GGACTACHVGGGGTWTCTAAT-3'). The PCR products separated by 2% agarose gel electrophoresis were extracted and purified by AP-GX-500 DNA gel extraction kit. After library establishment, the obtained products were sequenced on the MiSeq sequencing platform. The modified and installed sequences of each sample were unified into the Gree...
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