Inhibition on Prion Replication by Ethanol Crude Extracts from Inonotus obliquus
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Abstract:
To optimize the extraction process of the ethanol crude extracts from Inonotus obliquus and preliminarily study its effect on prion replication. The optimum extraction condition of ethanol crude extracts from Inonotus obliquus were employed by alcohol extraction and orthogonal test. To evaluate the cell safety of the ethanol crude extracts from Inonotus obliquus, CCK-8 kit was used to determine the safe concentration. According to the results of safety evaluation, Western blot was used to detect the effect of the ethanol crude extracts from Inonotus obliquus on prion replication. The antioxidant activity of ethanol crude extracts from Inonotus obliquus was studied by measuring hydrogen peroxide and antioxidant factors. The optimal extraction conditions were as follow: the ratio of solid to liquid was (Inonotus obliquus power: alcohol) 1:10 (g/mL), the volume fraction of ethanol was 75%, the extraction time was 2 h, the extraction temperature was 80 ℃. At this condition, the yield of ethanol crude extracts from Inonotus obliquus was 6.32%. The maximum safe concentration range of ethanol crude extracts from Inonotus obliquus was 6.25×10-4 mg/mL, Under this condition, after 24 h treatment, prion replication were inhibited by the ethanol crude extracts from Inonotus obliquus (p<0.01), which could reduce the content of H2O2 and increase the content of SOD and antioxidant factor GCLM、GCLC，and it was statistically significant. The results showed that ethanol crude extracts from Inonotus obliquus had an inhibitoryeffect on prion replication, which may be related to its antioxidant activity.
