Transcriptome Profiling ofToxoplasma gondii-Infected Human Cerebromicrovascular Endothelial Cell Response to Treatment with Monensin
Transcriptome Profiling ofToxoplasma gondii-Infected Human Cerebromicrovascular Endothelial Cell Response to Treatment with Monensin
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DOI：
10.3390/microorganisms8060842
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发表时间：
2020-06-01
期刊：
影响因子：
4.5
通讯作者：
Elsheikha, Hany M.
中科院分区：
文献类型：
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作者：
Harun, Mohammad S. R.;Taylor, Mica;Elsheikha, Hany M.
弓形虫与血脑屏障(BBB)内皮细胞的相互作用是脑弓形虫病进展的中心。在目前的工作中，我们检验了一种假设，即单价离子载体莫能菌素抑制Wnt途径信号转导会减少Ft的生长。感染人脑微血管内皮细胞或小胶质细胞。莫能菌素(一种Wnt信号抑制剂)对体外生长的抑制作用。用两种方法(硫代罗丹明B染色和镜检寄生虫计数)对弓形虫进行了检测。莫能菌素抑制T。对hBMECs的选择指数为8.48(50%细胞毒浓度[CC50]=5.17 mU M)。然而，莫能菌素对小胶质细胞的IC(50)为4.13微米M，SI=13.82(CC50=57.08微米)，表明小胶质细胞对莫能菌素的敏感性较低。它的效果是。利用体外人血脑屏障模型，通过跨内皮细胞电阻(TEER)测定来评价血脑屏障的完整性。结果表明，该方法具有较好的抗菌活性。弓形虫感染可显著降低人脐静脉内皮细胞的TEER抗性，0.1 mU M莫能菌素处理细胞后，细胞对TEER的抗性得以恢复，这可能与莫能菌素的抗寄生虫活性有关。我们还研究了0.1 mU莫能菌素对hBMECs整体基因表达的宿主靶向性影响。淋病感染。用莫能菌素处理hBMECs并没有显著影响Wnt信号通路相关基因的表达，这表明尽管抑制Wnt信号通路并没有起到明显的INT作用。感染人脐静脉内皮细胞后，莫能菌素仍能有效地抑制其生长。贡地。相反，莫能菌素处理下调了与类固醇、胆固醇和蛋白质生物合成相关的通路及其在内质网和高尔基体之间的运输，并下调了与细胞周期和DNA合成修复机制相关的通路。这些结果为研究莫能菌素的寄主调控作用提供了新的视角。弓形虫感染，值得进一步调查。
Central to the progression of cerebral toxoplasmosis is the interaction ofToxoplasma gondiiwith the blood-brain barrier (BBB) endothelial cells. In the present work, we tested the hypothesis that inhibition of Wnt pathway signalling by the monovalent ionophore monensin reduces the growth ofT. gondiiinfecting human brain microvascular endothelial cells (hBMECs) or microglial cells. The anti-parasitic effect of monensin (a Wnt signalling inhibitor) on the in vitro growth ofT. gondiitachyzoites was investigated using two methods (Sulforhodamine B staining and microscopic parasite counting). The monensin inhibitedT. gondiigrowth (50% inhibitory concentration [IC50] = 0.61 mu M) with a selective index = 8.48 when tested against hBMECs (50% cytotoxic concentration [CC50] = 5.17 mu M). However, IC(50)of monensin was 4.13 mu M with a SI = 13.82 when tested against microglia cells (CC50= 57.08 mu M), suggesting less sensitivity of microglia cells to monensin treatment. The effect ofT. gondiion the integrity of the BBB was assessed by the transendothelial electrical resistance (TEER) assay using an in vitro human BBB model. The results showed thatT. gondiiinfection significantly decreased hBMECs' TEER resistance, which was rescued when cells were treated with 0.1 mu M monensin, probably due to the anti-parasitic activity of monensin. We also investigated the host-targeted effects of 0.1 mu M monensin on global gene expression in hBMECs with or withoutT. gondiiinfection. Treatment of hBMECs with monensin did not significantly influence the expression of genes involved in the Wnt signalling pathway, suggesting that although inhibition of the Wnt signalling pathway did not play a significant role inT. gondiiinfection of hBMECs, monensin was still effective in limiting the growth ofT. gondii. On the contrary, monensin treatment downregulated pathways related to steroids, cholesterol and protein biosynthesis and their transport between endoplasmic reticulum and Golgi apparatus, and deregulated pathways related to cell cycle and DNA synthesis and repair mechanisms. These results provide new insight into the host-modulatory effect of monensin duringT. gondiiinfection, which merits further investigation.