Application of angelica protein to preparation of medicine for protecting against acute liver damage
A liver injury and protein technology, applied in the application field of angelica protein in the preparation of liver injury protection drugs, to achieve the effect of reducing injury, reducing the activity of GST and T-AOC, and improving the necrosis of liver cells
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Embodiment 1
[0031] Example 1: Purification of Angelica ASPR protein
[0032] Soak the angelica cut into small pieces with 10 times the volume of 0.05 mol / L Tris-HCl buffer (pH 8.0), let stand at 4°C overnight for about 15 h, filter the residue with 4 layers of gauze the next day, and filter the filtrate at 12000 rpm, 4 Centrifuge at ℃ for 10 min, 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 the precipitated protein is collected and dissolved in 2 times the volume of 0.05 mol / L Tris- In HCl buffer (pH 8.0), the solution of ammonium sulfate precipitation was loaded onto a Sephadex G-50 chromatography column fully equilibrated with equilibration solution (0.05 mol / L, pH 8.0 Tris-HCl buffer). The equilibration solution was eluted and the second eluted peak was collected, and the purity was checked by SDS-PAGE electrophoresis.
[0033] Experime...
Embodiment 2
[0035] Example 2: Proliferative effect of Angelica ASPR protein on L-02 cells
[0036] The effect of Angelica ASPR protein on the survival rate of L-02 cells was detected by MTT assay. Cell viability was calculated according to the following formula.
[0037] Cell survival rate (%)=(OD value of test 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 in a concentration-dependent manner, such as figure 2 As shown, the survival rate of L-02 cells increased with the concentration at 0.063-0.50 mg / mLL. At 0.5-0.625 mg / mLL, the survival rate of L-02 cells decreased with the increase of concentration, and at the dose of 0.5 mg / mLL, Angelica ASPR protein could increase the proliferation rate of L-02 to 163.14%.
Embodiment 3
[0040] Example 3: For CCl 4 Dose Exploring for Preventive Effects of Induced Acute Liver Injury
[0041] 1. Experimental animals and grouping
[0042] Male Kunming mice, purchased from Wu's Animals, weighed 20 ± 2 g. The animals were fed normally for three days before the experiment. The mice were randomly divided into 5 groups with 5 mice in each group. The groups and their treatments were as follows: normal group (CON) and liver injury group (CCl 4 ) were injected with 0.5 mL of normal saline for 3 days, and 2 hours after the last administration, the CON group was injected with 0.2 mL of olive oil, CCl 4 The group was injected with 0.1% CCl 4 Solution 0.2 mL (20 mL / kg); protein dosing group [ASPR1 (0.5 mg / mL) + CCl 4 , ASPR2 (1.25mg / mL) + CCl 4 , ASPR3 (2.0 mg / mL + CCl 4 )] were continuously injected with 0.5 mL of low, medium and high protein solution for 3 d, and 0.1% CCl was injected 2 h after the last administration 4 Olive oil solution 0.2 mL (20 mg / kg).
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