title: Inhibitory effects and analysis of RNA interference on thioredoxin glutathione reductase expression in Schistosoma japonicum.
abstract: Schistosomes infect around 280 million people worldwide. The worms survive in the veins of the final host, where thioredoxin glutathione reductase (TGR) activity helps the parasites to survive in the aerobic environment. In the present study, we synthesized 4 small interfering RNAs (siRNA S1, S2, S3, and S4) targeting the Schistosoma japonicum (Sj) TGR gene and used them to knockdown the TGR gene. The knockdown effects of the siRNAs on SjTGR, and the thioredoxin reductase (TrxR) activity of SjTGR, were evaluated in vitro. The results of transfection with the siRNAs via the soaking method in vitro were confirmed by flow cytometry. S2 siRNA at a final concentration of 200 nM partially inhibited the expression of SjTGR at both the transcript and protein levels in vitro. TrxR-activity was lower in worms in the S2 siRNA-treated group compared with the control groups. Further analysis revealed that purified recombinant SjTGR could remove oxygen free radicals but not H(2)O(2) directly, which may explain the incomplete effects of RNA interference on SjTGR. The results of this study indicate that SjTGR may play an important role in the clearance of oxygen free radicals and protection of S. japonicum parasites against oxidative damage.
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, P.R. China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Han
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Fu
authors.forename: Yang
authors.initials: Y
authors.lastname: Hong
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: Hongxiao
authors.initials: H
authors.lastname: Han
authors.forename: Ke
authors.initials: K
authors.lastname: Lu
authors.forename: Jianmei
authors.initials: J
authors.lastname: Yang
authors.forename: Xiangrui
authors.initials: X
authors.lastname: Li
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Lin
mesh_terms: D000818:Animals; D004278:DNA, Superhelical / Q000502:physiology; D055785:Gene Knockdown Techniques; D006861:Hydrogen Peroxide / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000502:physiology; D011817:Rabbits; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics; D012908:Snails; D014158:Transcription, Genetic; D014162:Transfection / Q000592:standards
chemical_list: D004278:DNA, Superhelical; D009097:Multienzyme Complexes; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D006861:Hydrogen Peroxide; D009247:NADH, NADPH Oxidoreductases; C466433:thioredoxin glutathione reductase