动物造模 z-bio 2022-06-09 646

　　OBJECTIVE: To observe the expression of Sjp40 in infected New Zealand white rabbits with hepatic sediment eggs and the formation of granuloma lesions, and to perform immunolocalization.

　　Methods: New Zealand white rabbits were infected with Schistosoma japonicum cercariae, and the livers of the uninfected group, 29 dpi group and 45 dpi group were collected, and the total RNA of each group was extracted by Trizol method, using Schistosoma japonicum glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as the internal reference , Taqman probe qRT-PCR was used to detect the expression of Sjp40 mRNA; the total liver protein was extracted from each group, and the anti-Sjp40-McAb 9G7 and anti-Toxoplasma tSAG1-McAb were purified by ammonium sulfate salting out method and Protein G immunoaffinity column chromatography. Y3A8 (control antibody), western blot was used to detect the expression of Sjp40 protein; liver paraffin sections were prepared, and HE staining was used to observe the formation and development of hepatic egg granulomas. The immunofluorescence technique was further used to detect the expression of Sjp40 in liver deposited eggs and granuloma tissue. positioning.

　　Results: The level of Sjp40 mRNA in the 45 dpi liver deposited eggs of Schistosoma japonicum-infected rabbits with acute hepatic granuloma lesions was significantly higher than that in the 29 dpi immature eggs deposited in the liver without egg granuloma nodules (P<0.05). , Westernb blot confirmed the expression of Sjp40 protein in 29 dpi and 45 dpi rabbit liver. Immunofluorescence showed that Sjp40 was localized in immature eggs in the liver of rabbits at 29 dpi, while at 45 dpi, a large number of mature eggs and surrounding granulomas were deposited in the liver of infected rabbits.

　　CONCLUSION: The transcript level of Sjp40 in the eggs deposited in the liver of Schistosoma japonicum-infected rabbits increases significantly with the maturity of the eggs. During the acute granulomatous stage (45dpi) of infected rabbits, the eggs spread to the surrounding granuloma tissue, suggesting that This molecule plays an important role in the formation and development of schistosomiasis granulomas.