Application of a kind of angelica protein in the preparation of liver injury protection medicine
A liver injury prevention drug technology, applied in the application field of angelica protein in the preparation of liver injury protection drugs, to achieve the effect of improving liver cell necrosis, reducing damage, and reducing liver MDA levels
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Embodiment 1
[0031] Embodiment 1: the purification of Angelica ASPR protein
[0032] Soak the Angelica sinensis cut into small pieces in 10 times the volume of 0.05 mol / L Tris-HCl buffer solution (pH 8.0), let it stand overnight at 4°C for about 15 hours, filter the residue with 4 layers of gauze the next day, and filter the filtrate at 12,000 rpm, 4 Centrifuge at ℃ for 10 minutes, and the obtained supernatant is the crude extract of angelica protein; the crude protein solution is subjected to one-step precipitation with ammonium sulfate (0-80%), precipitated overnight and collects the precipitated protein, which is dissolved in 2 times the volume of 0.05 mol / L Tris- In HCl buffer solution (pH 8.0), the ammonium sulfate precipitated solution was loaded onto the Sephadex G-50 chromatography column fully equilibrated with the equilibrium solution (0.05 mol / L, pH 8.0 Tris-HCl buffer solution), and the The balance solution was eluted and the second eluted peak was collected, and the purity was...
Embodiment 2
[0035] Example 2: Proliferative effect of Angelica ASPR protein on L-02 cells
[0036] MTT method was used to detect the effect of Angelica ASPR protein on the survival rate of L-02 cells. Cell viability was calculated according to the following formula.
[0037] Cell survival rate (%)=(OD value of measurement group-OD value of blank group) / (OD value of control group-OD value of blank group)×100.
[0038] Experimental results:
[0039] Angelica ASPR protein can effectively promote the proliferation of L-02 cells, and has a concentration-dependent relationship, such as figure 2 As shown, when the concentration was 0.063-0.50 mg / mLL, the survival rate of L-02 cells increased with the increase of the concentration. At 0.5-0.625 mg / mLL, the survival rate of L-02 cells decreased with the increase of the concentration, and at the dose of 0.5 mg / mLL, Angelica ASPR protein could make the proliferation rate of L-02 reach 163.14%.
Embodiment 3
[0040]Embodiment 3: to CCl 4 Dose-finding for the prevention of induced acute liver injury
[0041] 1. Experimental animals and grouping
[0042] Male Kunming mice, purchased from Wu's animals, weighing 20 ± 2 g. Normal feeding for three days before the experiment. The mice were randomly divided into 5 groups, 5 mice in each group, each group and its treatment were as follows: normal group (CON) and liver injury group (CCl 4 ) were injected with 0.5 mL of normal saline for 3 consecutive days, and 2 hours after the last administration, the CON group was injected with 0.2 mL of olive oil, CCl 4 Group injected with 0.1% CCl 4 Solution 0.2 mL (20 mL / kg); Protein plus drug group [ASPR1 (0.5mg / mL) + CCl 4 , ASPR2 (1.25mg / mL) + CCl 4 , ASPR3 (2.0mg / mL + CCl 4 )] Continuous injection of 0.5 mL of low, medium and high protein solution for 3 days respectively, 2 hours after the last administration, and then injection of 0.1% CCl 4 Olive oil solution 0.2 mL (20 mg / kg).
[0043] ...
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