氨氮、镉胁迫下对虾Hippo通路对肠道恢复的响应

Response of Hippo Pathway to Intestinal Injury Recovery in Pacific White Shrimp Litopenaeus vannamei Exposed to Ammonia Nitrogen and Cadmium Stresses

为探明在氨氮和镉分别胁迫下凡纳滨对虾Hippo信号通路对肠道损伤恢复的响应,在(3.99±0.50)g对虾胁迫阶段(0、24、48 h),以及恢复阶段的3、6、12、24、48 h检测Hippo信号通路中部分基因在肠道组织中的表达情况,同时进行肠道组织学观察。结果显示:在氨氮胁迫48 h后恢复阶段,与对照组相比,Wts、Yki、TEAD基因表达量在恢复阶段的6 h极显著升高(P<0.01);TEAD基因表达量在恢复阶段的12 h显著升高(P<0.05);Hpo基因表达量除在恢复阶段的3 h无显著差异外,其余阶段均极显著降低(P<0.01);Wts基因表达量在恢复阶段的3 h和恢复阶段的24 h极显著降低(P<0.01)且在恢复阶段的12 h和48 h显著降低(P<0.05)。在镉胁迫后,与对照组相比,Wts基因表达量在胁迫后24 h和48 h均极显著降低(P<0.01);Hpo和Wts基因表达量除Hpo基因表达量在恢复阶段的6 h和Wts基因表达量在恢复阶段的48 h无显著差异外,其余时间均极显著降低(P<0.01);Yki基因表达量在恢复阶段的6、12 h和24 h均极显著升高(P<0.01);TEAD基因表达量在恢复阶段的6 h极显著升高(P<0.01)。同时,肠道组织学分析显示,对虾肠道分别在氨氮胁迫后的48 h和镉胁迫后24 h肠绒毛完全脱落,在恢复阶段的24 h肠绒毛有明显的恢复生长迹象。试验结果表明,Hippo信号通路参与凡纳滨对虾氨氮和镉单因素胁迫下肠绒毛恢复与再生过程的调控。

In order to explore the response of Hippo signaling pathway to intestinal injury recovery in Pacific white shrimp Litopenaeus vannamei under single-factor ammonia nitrogen or cadmium stress,the gene expressions of Hippo signaling pathway was examined in intestinal tissues of Pacific white shrimp with body weight of(3.99±0.50) g exposed to ammonia nitrogen or cadmiumstresses at 0 h,24 h and 48 h during the stress phase,and at 3 h,6 h,12 h,24 h and 48 h during the recovery phase,and intestinal histology was observed at the same time.The results showed that there was very significantly higher relative expression levels of Wts,Yki and TEAD genes at 6 h in the recovery stage after ammonia nitrogen stress compared with the control group(P<0.01),with significantly higher expression level of TEAD gene at 12 h in the recovery stage(P<0.05) and without significant difference in Hpo gene expression level,and the other stages was very significantly lower(P<0.01) in the recovery stage except for 3 h.Wts gene was found to be very significantly lower at 3 h and 24 h in the recovery stage(P<0.01) and significantly lower at 12 h and 48 h in the recovery stage(P<0.05).Wts gene was significantly lower at 24 h and 48 h in the shrimp exposed to cadmium stress compared with the control group(P<0.01).There were very significantly lower Hpo and Wts genes expression levels in the shrimp exposed to cadmium stress(P<0.01) except for Hpo gene at 6 h in the recovery stage and Wts gene at 48 h in the recovery stage.Yki gene was very significantly lower at 6 h,12 h and 24 h in the recovery stage(P<0.01);TEAD gene was very significantly higher at 6 h in the recovery stage(P<0.01).Intestinal histological observation showed that the intestinal villus of the shrimp was completely shed at 48 h in the ammonia-nitrogen stress stage and 24 h in the cadmium stress stage,and that the intestinal villi showed obvious signs of recovery and growth at 24 h in the recovery stage.The findings indicated that the Hippo signaling pathway was involved in the regulation of the recovery and regeneration of intestinal villus under the single-factor stress of ammonia nitrogen and cadmium in Pacific white shrimp.