[The Mechanism of Echinococcus Granulosus Sensu Stricto Antigen B to Protect Immune Thrombocytopenia Mouse Model by Influencing Autophagy].

OBJECTIVE

To investigate the mechanism of natural antigen B (nAgB) to protect Immune thrombocytopenia (ITP) mouse model by influencing autophagy.

METHODS

Twenty-eight female BALB/c mice aged 8-10 weeks were randomly divided into four groups. 7 mice of each group were immunized intraperitoneally, the control group was treated with PBS as the control group; ITP group was treated with anti-CD41 monoclonal antibody (anti-CD41Ab) only; nAgB group was treated with nAgB intraperitoneal injection for 5d; nAgB+ITP group was treated with nAgB intraperitoneal injection for 5d, then treated with anti-CD41 Ab. The peripheral blood platelet count in each group was tested; and the spleen and liver should be isolated and weighed, the organ index was calculated; qRT-PCR was used to detect spleen microtubule-associated protein 1 light chain 3 ( LC3), p62, Beclin-1 mRNA expression levels. Western blot was used to detect the protein expression level of spleen LC3Ⅱ/LC3Ⅰ, p62, Beclin-1.

RESULTS

Compared with the control group, mice in the ITP group showed a significant decrease in blood PLT count ［(102.1±17.9)×109/L vs (485.4±185.2)×109/L, P <0.01］, a significant increase in spleen index (P <0.01), mice in the nAgB group showed a significant increase in blood PLT count, rising to (1051±127.6)×109/L on the 3 day after modeling. Compared with the ITP group, mice in the nAgB+ITP group showed a significant increase in PLT count on the 1 day of anti-CD41 Ab administration ［(428.6±131.6)×109/L vs (102.1±17.9)×109/L, P <0.05］, however, the spleen index was significantly decreased (P <0.05). qRT-PCR and Western blot results showed that compared with the control group, the mRNA and protein expression levels of spleen LC3, p62 and Beclin-1 were increased in the ITP group of mice (P <0.05, P <0.01). Compared with the ITP group, the nAgB+ITP group could significantly decrease mRNA levels of spleen LC3, p62 and Beclin-1 (P <0.05, P <0.01), and also significantly decrease the protein expression levels of LC3Ⅱ/LC3Ⅰ, p62 and Beclin-1 (P <0.05, P <0.01).

CONCLUSION

nAgB inhibits the transcription and expression levels of autophagy-related genes and regulates immune intolerance, thereby protecting ITP mouse models.