Applications of EGCG in preparing drugs or health food for improving or treating endogenous endotoxemia
A technology for endotoxemia and health food, applied in the fields of biology and pharmacy, can solve the problems of low bioavailability of EGCG, and achieve the effect of improving endotoxemia and simple method
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Embodiment 1
[0024] Example 1 Grouping of experimental animals and establishment of endogenous endotoxin model induced by high-fat diet
[0025] Experimental materials and animals: Epigallocatechin gallate (EGCG) was purchased from Shanghai Yuanye Biotechnology Co., Ltd., item number S27863, lipopolysaccharide (LPS) assay kit was purchased from Beijing Huaying Institute of Biotechnology, male C57BL / 6J was purchased from the Experimental Animal Center of Nankai University, high-fat diet D12492 and low-fat control diet D12450J were both purified feeds, and were purchased from Jiangsu Synergy Pharmaceutical Bioengineering Co., Ltd.
[0026] experimental method:
[0027] (1) Place 30 6-week-old C57BL / 6 mice in an environment with a room temperature of 20-25°C and a 12h light / 12h dark cycle for one week. After adaptive feeding, the mice were randomly divided into 3 groups: normal diet For the control group (NC), high-fat diet model control group (HFD), and EGCG intervention group (HFD+EGCG), E...
Embodiment 2E
[0030] Example 2 Effect of EGCG on chylomicron synthesis marker gene ApoB48
[0031] Experimental materials: TRIzol was purchased from Ambion Company of the United States, and the reverse transcription kit LunaScript SuperMix Kit and qPCR kit Luna Universal qPCR Master Mix were purchased from NEB Company of the United States.
[0032] Experimental method: the total RNA in the ileum tissue was extracted with Trizol reagent, the purity of the RNA was detected with a Nanophotometer, and a 1% (w / v) agarose gel was prepared to detect its integrity. cDNA was synthesized using a reverse transcription kit and stored at -80°C for future use.
[0033] The expression level of mRNA was analyzed by qPCR, qPCR system (20 μL): Luna Universal qPCR Mix 10 μL, cDNA template 4 μL, upstream primer (10 μM) 0.5 μL, downstream primer (10 μM) 0.5 μL, ddHO supplemented to 20 μL. The PCR reaction program was: pre-denaturation at 95°C for 60s, denaturation at 95°C for 15s, extension at 60°C for 30s, 40...
Embodiment 3
[0036] Example 3 Effect of EGCG on the synthesis of chylomicrons in Caco2 cells
[0037] Experimental materials: oleic acid was purchased from Sigma, USA, item number 01008
[0038] experimental method:
[0039] (1) Establishment and verification of the Caco2 cell monolayer model: use DMEM high-glucose medium containing 10% fetal bovine serum, 1% penicillin-streptomycin double antibody solution and 1% glutamine at 37°C, containing 5 %CO 2 Caco2 cells were cultured under the environment. In a Transwell chamber (0.4 μm, 1.12 cm) suspended in a 12-well plate 2 ) The seeding density in the upper chamber was 1×10 6 0.5ml of Caco-2 cell suspension per ml, then put 1.5ml of blank culture medium in the lower chamber, and culture it for 14-21 days to make it differentiate, thereby simulating the tight junction protein structure of small intestinal epithelial cells, when the transmembrane resistance value When >600Ω, the cell monolayer can be considered dense and complete.
[0040] ...
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