PDF(Pubmed)
Abstract：
Endoplasmic reticulum stress (ERS) and oxidative stress (OS) are adaptive responses of the body to stressor stimulation. Although it has been verified that Trichinella spiralis (T. spiralis) can induce ERS and OS in the host, their association is still unclear. Therefore, this study explored whether T. spiralis-secreted serpin-type serine protease inhibitor (TsAdSPI) is involved in regulating the relationship between ERS and OS in the host intestine. In this study, mice jejunum and porcine small intestinal epithelial cells (IECs) were detected using qPCR, western blotting, immunohistochemistry (IHC), immunofluorescence (IF), and detection kits. The results showed that ERS- and OS-related indexes changed significantly after TsAdSPI stimulation, and Bip was located in IECs, indicating that TsAdSPI could induce ERS and OS in IECs. After the use of an ERS inhibitor, OS-related indexes were inhibited, suggesting that TsAdSPI-induced OS depends on ERS. When the three ERS signalling pathways, ATF6, IRE1, and PERK, were sequentially suppressed, OS was only regulated by the PERK pathway, and the PERK-eif2α-CHOP-ERO1α axis played a key role. Similarly, the expression of ERS-related indexes and the level of intracellular Ca2+ were inhibited after adding the OS inhibitor, and the expression of ERS-related indexes decreased significantly after inhibiting calcium transfer. This finding indicated that TsAdSPI-induced OS could affect ERS by promoting Ca2+ efflux from the endoplasmic reticulum. The detection of the ERS and OS sequences revealed that OS occurred before ERS. Finally, changes in apoptosis-related indexes were detected, and the results indicated that TsAdSPI-induced ERS and OS could regulate IEC apoptosis. In conclusion, TsAdSPI induced OS after entering IECs, OS promoted ERS by enhancing Ca2+ efflux, and ERS subsequently strengthened OS by activating the PERK-eif2α-CHOP-ERO1α axis. ERS and OS induced by TsAdSPI synergistically promoted IEC apoptosis. This study provides a foundation for exploring the invasion mechanism of T. spiralis and the pathogenesis of host intestinal dysfunction after invasion.
摘要：
内质网应激(ERS)和氧化应激(OS)是机体对应激源刺激的适应性反应。虽然已经证实旋毛虫(T.螺旋）可以在宿主中诱导ERS和OS，他们的关系还不清楚。因此,这项研究探讨了螺旋T.serpin型丝氨酸蛋白酶抑制剂（TsAdSPI）是否参与调节宿主肠道中ERS和OS之间的关系。在这项研究中,使用qPCR检测小鼠空肠和猪小肠上皮细胞（IECs），西方印迹,免疫组织化学（IHC），免疫荧光（IF），和检测试剂盒。结果显示,TsAdSPI刺激后ERS和OS相关指标发生显著变化,Bip位于IEC,表明TsAdSPI可以诱导IEC中的ERS和OS。使用ERS抑制剂后，OS相关索引被抑制，表明TsAdSPI诱导的OS依赖于ERS。当三种ERS信号通路，ATF6、IRE1和PERK,被依次压制，OS仅受PERK通路调节,PERK-eif2α-CHOP-ERO1α轴发挥了关键作用。同样，加入OS抑制剂后,ERS相关指标的表达和细胞内Ca2+水平均受到抑制,抑制钙转移后,ERS相关指标的表达明显下降。这一发现表明TsAdSPI诱导的OS可以通过促进Ca2+从内质网流出来影响ERS。对ERS和OS序列的检测显示OS发生在ERS之前。最后,检测细胞凋亡相关指标的变化，结果表明，TsAdSPI诱导的ERS和OS可以调节IEC凋亡。总之,TsAdSPI在进入IEC后诱导OS，OS通过增强Ca2+流出来促进ERS，ERS随后通过激活PERK-eif2α-CHOP-ERO1α轴来增强OS。TsAdSPI诱导的ERS和OS协同促进IEC凋亡。本研究为探索旋毛虫的侵袭机制及侵袭后宿主肠功能障碍的发病机制奠定了基础。
