The Inhibiting Effects Of The Recombinant Schistosoma Japonicum T2 RNase And SjE16 Proteins On HSC Activation

Objective To investigate the effects of recombinant Schistosoma japonicum T2 RNase(rT2 RNase)and SjE16(rSjE16)proteins on hepatic stellate cell(HSC)activation.Methods1.LX-2 cells were treated with the recombinant protein rT2 RNase and the expression level of alpha-smooth muscle actin(?-SMA)which is a characteristic marker of activated HSCs was assessed by Western Blot.Meanwhile,the effects of the rT2 RNase on the expression levels of the signal transduction molecule Smad4 and NF-?B were further observed.2.Total RNA was extracted from Schistosoma japonicum eggs and then reverse transcribed into cDNA.Based on the sequence from Pubmed(GenBank: FN320650.1),the full length sequence of Schistosoma japonicum E16 gene was obtained by PCR.The recombinant protein rSjE16 was acquired though the prokaryotic expression system.Then,LX-2 cells were treated with the fusion protein rSjE16,subsequently the cell viability was examined by MTT assay and the cell cycle was analyed by flow cytometry.The effects of rSjE16 on the expression of fibrotic genes and the inflammatory cytokines were detected with the methods of Western Blot or qRT-PCR.Results1.?-SMA expression was inhibited in LX-2 cells treated with rT2 RNase at various concentrations for different times.Meanwhile,rT2 RNase could induce the expression of PPAR?.rT2 RNase could also inhibit the up-regulation of ?-SMA in TGF-?1 treated LX-2 cells through the inhibition of Smad4.Moreover,rT2 RNase also reduced NF-?B expression and p-I?B? expression.2.The recombinant rSjE16 protein was expressed and purified successfully.The protein expression levels of ?-SMA and collagen?were significantly reduced by rSjE16 in LX-2 cells,while the expression levels of PPAR?,MMP2 and MMP9 were up-regulated.rSjE16 also increased the mRNA expression levels of some inflammatory factors such as IL-6 and IL-8.However,rSjE16 has no effect on cell proliferation and cell cycle of HSCs.Conclusions1.rT2 RNase suppressed the activation of HSCs through TGF-?1 and NF-?B signaling pathway.2.rSjE16 inhibited the activation of HSCs though regulating the expression levels of fibrotic genes and increased the generations of some inflammatory factors in HSCs.