Preliminary Study Of Characterization Of SjShh Gene And Function Of SjWnt1 Protein In Schistosoma Japonicum

Schistosomiasis caused by Schistosoma japonicum(S.japonicum)is a severe parasitic disease,which imposed considerable burden on human,domestic animal health and socioeconomic development.While a single drug treating of S.japonicum was easy to tolerance.Hedgedge(Hh)and Wnt signaling pathway are related to the development,and both of these affect the development of individual embryos and maintain tissue homeostasis.Inhibition of Sonic hedgehog(Shh)in hedgehog signaling pathway and wnt1 in wnt signaling pathway by siRNA interference,makde sure the function in S.japonicum.This could also provide ideas for the new target genes of S.japonicum.1?Characterization analysis and the relative mRNA expression levels of different stages and genders of S.japonicum.Using NCBI retrieval Shh EST and full sequence is obtained by RACE technique and named SjShh gene.We carried on the bioinformatics analysis,an open reading frame with 2145 bp.Sj Shh proteins are composed of two distinct domains,the amino-terminal ‘Hh' domain(HhN),and the carboxy-terminal ‘Hot' domain(HhC).The analysis of the transcriptional levels of Shh showed that SjShh gene plays an important role in the whole development stage of S.japonicum.The expression of mRNA levels at different developmental stages presented that SjShh expressed in all development stages of S.japonicum,and expressions level in schistosomulum stages were higher than other development stages,and 18-day-old schistosomulum reached maximum,suggesting that SjShh can regular development system.We also found that SjShh mRNA level in maldevelopment worm was higher than that of normal worms,which suggested dysplasia can cause Hh pathway transduction latency..2?Study on the function of SjShh protein in S.japonicumEffective and stable of small molecule Shh796 was successfully screened and the optimal interference concentration was 1.5OD/ml.Troughed inner interference in different stages of 14 day and 24 day,the results showed that the inhibition effect was significance;outer experiment of SjShh siRNA interference test has shown significance inhibition effect was in 72 hours.However,the SjShh siRNA interference had no significant effect on the morphology of the worms and no obvious impacts on the routine blood test of the host.Biochemical indicators of ALT,AST,and LDH were reduced;suggesting that interference of the Hh pathway may be reduced to injury of host liver.3?Study on the function of Sj Wnt protein in S.japonicum The results showed that the function of SjWnt1 protein was mainly related to the reproduction of worms.The SjWnt1 protein was mainly secreted in the lower layer of integument.Positive particles around the intestine,testis and distribution of testis were the most intensive.The expression of mRNA in SjWnt1 was down-regulated and the transcriptional level of SjWnt1 was decreased.The expression of mRNA in downstream pathway was also down-regulated,indicated that the SjWnt1 was involved in Wnt / ?-catenin,Wnt / PCP and Wnt / Ca2 + signaling pathways.In vitro siRNA interference after the parasite electron microscopy results: the cortex becomes swollen,the body was larger vacuoles,basement membrane dissolution,diffusion,body spine reduction,mitochondrial proliferation and accompanied by severe swelling,intestinal microvilli shedding,fracture SjWnt1 inhibition of S.japonicum surface damage,insect energy metabolism imbalance,the body's own steady-state affected.In this paper,the SjShh gene was obtained by Race,and the expression of SjShh gene in S.japonicum was determined by qPCR.In vivo and in vitro siRNA interference experiments of SjShh and SjWnt1,which provided a basis for understanding the functional role of SjShh and SjWnt1 in S.japonicum.