Department of Parasitology, Medical College, Zhengzhou University, Zhengzhou, 450052, China.
Parasitol Res. 2013 Dec;112(12):4113-20. doi: 10.1007/s00436-013-3602-1. Epub 2013 Sep 12.
The activation of Trichinella spiralis muscle larvae (ML) by exposure to intestinal contents or bile and the intestinal epithelial cells (IECs) themselves are two pivotal requirements for the in vitro larval invasion of IECs. However, it is yet unknown which genes are involved in the process of larval invasion. The purpose of the present study was to analyze the differentially expressed genes of T. spiralis larvae activated by bile and cultured with IECs by using real-time polymerase chain reaction. Ten T. spiralis genes encoded the proteins produced by the larvae after co-culture with IECs were selected. Compared with untreated ML, four genes were up-regulated in both bile-activated and cell-cultured larvae, including calcium-dependent secretion activator (Csa; 2.55- and 16.04-fold, respectively), multi cystatin-like domain protein precursor (Mcd; 4.36 and 52), serine protease (Sp; 2.03 and 20.02), and intermediate filament protein ifa-1 (Ifa 1; 2 and 3.31). The expression of two genes, enolase (Eno; 1.51) and ribosomal protein S6 kinase beta-1 (Rsk; 1.49), was up-regulated only in cell-cultured larvae, not in bile-activated larvae. The expression of secreted 5'-nucleotidase (5 N; 1.42) and putative serine protease (Psp; 1.41) was up-regulated in bile-activated larvae, but was not changed or down-regulated after cultured with IECs. ATP synthase F1, beta subunit (ATPase; 0.58 and 0.51) and serine protease precursor (Spp; 0.42 and 0.65) were down-regulated in both bile-activated and cell-cultured larvae. This study provide some differentially expressed genes among the untreated (normal), bile-activated and cell-cultured larvae of T. spiralis. The up-regulated genes might be related with the larval invasion of IECs, but their exact biological functions need to be further investigated. This study will be helpful to further elucidate the molecular mechanism of the invasion of IECs by T. spiralis larvae and to better understand the interaction between parasite and host enterocytes.
旋毛虫肌幼虫（ML）被暴露于肠内容物或胆汁以及肠上皮细胞（IECs）本身激活，这是幼虫体外入侵 IECs 的两个关键要求。然而，目前尚不清楚哪些基因参与了幼虫入侵的过程。本研究旨在通过实时聚合酶链反应分析用胆汁激活并用 IEC 培养的旋毛虫幼虫的差异表达基因。选择了 10 个旋毛虫幼虫基因，这些基因编码幼虫与 IEC 共培养后产生的蛋白质。与未经处理的 ML 相比，在胆汁激活和细胞培养的幼虫中，有 4 个基因上调，包括钙依赖性分泌激活剂（Csa；分别为 2.55 和 16.04 倍）、多半胱氨酸样域蛋白前体（Mcd；4.36 和 52）、丝氨酸蛋白酶（Sp；2.03 和 20.02）和中间丝蛋白 ifa-1（Ifa 1；2 和 3.31）。两个基因，烯醇酶（Eno；1.51）和核糖体蛋白 S6 激酶β-1（Rsk；1.49）的表达仅在细胞培养的幼虫中上调，而在胆汁激活的幼虫中没有上调。分泌的 5'-核苷酸酶（5N；1.42）和假定的丝氨酸蛋白酶（Psp；1.41）在胆汁激活的幼虫中上调，但在用 IEC 培养后没有变化或下调。ATP 合酶 F1，β亚基（ATPase；0.58 和 0.51）和丝氨酸蛋白酶前体（Spp；0.42 和 0.65）在胆汁激活和细胞培养的幼虫中均下调。本研究提供了旋毛虫未经处理（正常）、胆汁激活和细胞培养幼虫之间的一些差异表达基因。上调的基因可能与 IECs 的幼虫入侵有关，但它们的确切生物学功能需要进一步研究。本研究将有助于进一步阐明旋毛虫幼虫入侵 IECs 的分子机制，并更好地理解寄生虫与宿主肠细胞的相互作用。
