Use of lychee seed active ingredients in preparation of medicament for improving insulin resistance
An insulin resistance and drug technology, applied in the field of medicine, can solve the problems of improving peripheral and central insulin resistance effectively.
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Embodiment 1
[0043] PAS glycogen staining to observe the improvement effects of procyanidin A1 and procyanidin A2 on peripheral insulin resistance and central insulin resistance, respectively
[0044] S1: set Insulin resistance model group (simplified called, model group), including: HepG2 treated with 1 μmol·L -1 Peripheral insulin resistance model group formed after 48h of dexamethasone; and, treated with 2 μmol L -1 The central insulin resistance model group formed after dexamethasone treatment for 24 hours. Its specific cultivation operation steps are:
[0045] Establishment of peripheral insulin resistance model: take HepG2 cells in the logarithmic growth phase, trypsinize and adjust the cell density to 1×10 5 pcs mL -1 , inoculated in a 96-well plate, 100uL per well, placed in a 37°C, 5% CO2 incubator for 24h, after the cells adhered to the wall, discard the medium, and replace it with a serum-free medium for starvation induction for 12h, discard the medium, and use 1μmol· L -...
Embodiment 2
[0059] Effects of procyanidin A1 and / or procyanidin A2 on IRS-1, PI3K, Akt, GSK3β, Tau mRNA in HT22 cells
[0060] Real-time fluorescent quantitative PCR (Quantitative Real Time PCR) was used to verify the effects of procyanidin A1 and procyanidin A2 on IRS-1, PI3K, Akt, GSK3β, and Tau mRNA in HT22 cells. The specific operation steps were as follows:
[0061] 1. Cell treatment
[0062] Set up blank control group, central insulin resistance model group, pioglitazone group, litchi kernel mixed extract group, procyanidin A1 group and procyanidin A2 group. in,
[0063] Take HT22 cells in the logarithmic growth phase, trypsinize and adjust the cell density to 1×10 5 pcs mL -1 , seeded in a 6-well plate, 2 mL per well, at 37 ° C, 5% CO 2 In the incubator for 24 hours, after the cells adhered to the wall, replace the serum-free medium for starvation induction for 12 hours, discard the medium, and use 1 μmol L -1 After being treated with dexamethasone for 48 hours, the central insu...
Embodiment 3
[0096] Effects of procyanidin A1 and / or procyanidin A2 on the phosphorylation levels of IRS-1, PI3K, Akt, GSK3β and Tau total protein in HT22 cells
[0097] Western blot was used to verify the effects of procyanidin A1 and procyanidin A2, the active substances of litchi nucleus, on the phosphorylation levels of IRS-1, PI3K, Akt, GSK3β, and Tau proteins in HT22 cells. The specific experimental steps are as follows:
[0098] 1. Cell treatment is the same as in Example 2
[0099] 2. Protein extraction:
[0100] ①Discard the culture medium, wash with pre-cooled PBS for 3 times, each time for 5 minutes, prepare the lysate according to the ratio of RIRP strong lysate: protease inhibitor: phosphatase inhibitor = 8:1:1, add 200 μL of lysate to each well solution, lysed on ice for 30 min;
[0101] ②Scrape off the cells with a cell scraper, transfer them completely to a 1.5mL EP tube with a pipette tip, and lyse them by ultrasonication for 5sec;
[0102] ③ After centrifugation at 4°C...
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