Nematicidal activity of two monoterpenoids and SER-2 tyramine receptor of Caenorhabditis elegans
Nematicidal activity of two monoterpenoids and SER-2 tyramine receptor of Caenorhabditis elegans
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DOI：
10.1016/j.bcp.2009.11.002
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发表时间：
2010-04-01
影响因子：
5.8
通讯作者：
Enan, Essam
中科院分区：
文献类型：
--
作者：
Lei, Jason;Leser, Michael;Enan, Essam
建立了两种线虫（秀丽隐杆线虫和猪蛔虫）的体外培养模型，研究了3种单萜类化合物（麝香草酚、香芹酚和对伞花烃）的杀线虫活性。麝香草酚和香芹酚对两种线虫的毒力测定结果表明，麝香草酚和香芹酚对两种线虫的毒力均高于香芹酚。本研究的目的是通过使用转染了C.本研究旨在建立一个基于细胞的高通量抗蠕虫药物筛选平台。SER-2在转染细胞中的表达和功能性首先通过绿色荧光蛋白标记、竞争性受体结合、细胞内环AMP和细胞内钙[Ca 2 +]（i）动员测定来证实。然后测试百里酚和香芹酚，并证明在[Ca 2 +]（i）中对SER-2的酪胺活化脱敏中与TyrR相互作用;动员测定，以及在受体内化测定中将SER-2从膜易位到细胞质中。在表达具有S210 A/S214 A双突变的突变体SER-2的细胞中，百里酚和香芹酚的受体内化活性被显著阻断，从而证实了相互作用的特异性。总之，目前的研究表明，百里酚和香芹酚的杀线虫活性可能是通过TyrR介导的，因为这两种化合物可以在表达野生型而不是突变型SER-2的细胞中触发受体下游的信号级联。TyrR表达细胞系统可能被证明是一个很好的筛选平台，用于开发新的抗蠕虫化合物，可以克服寄生虫的耐药性，特别是当这些化学品与商业药物结合使用。(C)2009 Elsevier Inc. All rights reserved.
In vitro cultures of two nematodes (Caenorhabditis elegans and Ascaris suum) were established to study the nematicidal activity of three monoterpenoids (thymol, carvacrol and p-cymene). Toxicity of thymol and carvacrol was found for the two nematodes tested. The study was then aimed to address whether nematode tyramine receptor (TyrR) could interact with the two compounds by using HEK293 mammalian cells transfected with a C. elegans TyrR (ser-2) sequence, in hope of developing a high-throughput cell-based platform for future screening of new antihelminthic compounds. SER-2 expression and functionality in the transfected cells was first confirmed by green fluorescent protein tagging, competitive receptor binding, intracellular cyclic AMP, and intracellular calcium [Ca2+](i) mobilization assays. Thymol and carvacrol were then tested and demonstrated to interact with TyrR in desensitizing SER-2 for tyramine activation in [Ca2+](i); mobilization assay, and in translocating SER-2 from membrane to cytoplasm in receptor internalization assay. Receptor internalization activity of thymol and carvacrol was significantly blocked in cells expressing mutant SER-2 with the S210A/S214A double mutations, thus confirming specificity of the interactions. In summary, the current study showed that the nematicidal activity of thymol and carvacrol might be mediated through TyrR as the two compounds could trigger the signaling cascade downstream from the receptor in cells expressing wild-type but not a mutant SER-2. The TyrR-expressing cell system may prove to be a good screening platform for developing new antihelmintic compounds that may overcome parasite drug resistance, especially when such chemicals are used in combination with commercial drugs. (C) 2009 Elsevier Inc. All rights reserved.