Identification of differentially expressed genes in Oncomelania hupensis liver induced by sanguinarine / 中草药

ABSTRACT

Objective: To identify the differentially expressed genes in Oncomelania hupensis liver induced by sanguinarine. Methods: The median lethal concentration (LC50) of sanguinarine for O. hupensis was determined by concentration gradient method. Livers were isolated from live O. hupensis after being treated by 80% LC50 of sanguinarine or clean water. Then the mRNA of livers was purified and used for RT-PCR. The differentially expressed genes were acquired by suppression subtractive hybridization (SSH) and amplified by nested PCR before cloned into pMD-18T vecter. Positive clones were identified using PCR and DNA sequencing analyses. Then the further study was accomplished by bioinformatic analysis with blastx in NCBI. Results: The LC50 of sanguinarine for O. hupensis was 7.5 mg/L. The differentially expressed sequences were between 150 and 450 bp. After blastx homological analysis, the up-regulated genes were obtained as collagen α-4 (VI) chain, collagen α-5 (VI) chain, 40S ribosomal protein S19, hypothetical protein, kelch-like protein, and granulin-like protein. The down-regulated genes were obtained as β-tubulin, α-amylase 1, large subunit ribosomal protein L23e, chitotriosidase-1, Chit3 protein, hypothetical protein, mitochondrial aldehyde dehydrogenase, peptidoglycan recognition protein, eukaryotic translation elongation factor 1A, ferritin, disintegrin, and metalloproteinase with thrombospondin motifs 3, lysozyme 1, endo-1, 2 (4)-β-glucanase 1, and hemocyanin α-4D-subunit. The functions of these genes encoding proteins were related to material transportation, protein synthesis, proliferation induction, nutrition, anti-oxidation, anti-inflamation, respiration, signal transduction, and so on. Conclusion: Sanguinarine could cause the significant changes of gene expression in O. hupensis liver.