Badi Sara Ahmadi, Motahhary Atiyyeh, Bahramali Golnaz, Masoumi Morteza, Khalili Seyedeh Fatemeh Sadati, Ebrahimzadeh Nayereh, Nouri Pegah, Rahimi Ayoub, Masotti Andrea, Moshiri Arfa, Siadat Seyed Davar
Microbiology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
J Diabetes Metab Disord. 2020 Apr 22;19(1):415-422. doi: 10.1007/s40200-020-00522-3. eCollection 2020 Jun.
The intestine has substantial role in cholesterol homeostasis due to the presence of various cholesterol transporters and gut microbiota. spp. are important members of gut microbiota that employ outer membrane vesicles (OMVs) to interact with host. In this regard, we evaluated the effect of and related OMVs on the gene expression of important cholesterol transporters, Niemann-Pick C1-Like 1 (), ATP-binding cassette (), and liver X receptors (s) in Caco-2 cells.
OMVs were isolated from overnight brain heart infusion (BHI) broth of bacterial standard strains using deoxycholate and assessed by Scanning electron microscopy (SEM). The relative change in genes expression was assessed by Quantitative reverse transcription PCR (RT-qPCR) based on SYBR Green and 2 method in Caco-2 cells that were treated with bacteria and OMVs. Data were statistically analyzed with GraphPad Prism software. Finally, pathway enrichment based on the studied genes was performed using Cytoscape plugin ClueGO.
( = 0.002) and ( = 0.001) significantly reduced gene expression in Caco-2 cells. Interestingly, transcripts were significantly increased by both OMVs( = 0.04) ( = 0.01). Also, was significantly down regulated by ( = 0.02) ClueGO analysis on the studied genes demonstrated several functional groups which involve in lipid and cholesterol metabolism.
The opposite effect of , and related OMVs on the gene expression was observed in Caco-2 cells. Interestingly, these effects partially were in line with the alternation of s expression. However, based on pathway enrichment analysis, further molecular investigations are required to elaborate in details the specific association between spp. and OMVs with regulation of cholesterol signaling pathways including cholesterol transport, lipid storage, lipid homeostasis and cholesterol homeostasis.
由于存在各种胆固醇转运蛋白和肠道微生物群，肠道在胆固醇稳态中发挥着重要作用。[具体菌种名称]是肠道微生物群的重要成员，它们利用外膜囊泡（OMV）与宿主相互作用。在这方面，我们评估了[具体菌种名称]及其相关OMV对Caco-2细胞中重要胆固醇转运蛋白尼曼-匹克C1样1蛋白（NPC1L1）、ATP结合盒转运蛋白（ABCA1）和肝脏X受体（LXRs）基因表达的影响。
使用脱氧胆酸盐从细菌标准菌株的过夜脑心浸液（BHI）肉汤中分离OMV，并通过扫描电子显微镜（SEM）进行评估。基于SYBR Green和2-△△CT法，通过定量逆转录PCR（RT-qPCR）评估用细菌和OMV处理的Caco-2细胞中基因表达的相对变化。数据用GraphPad Prism软件进行统计分析。最后，使用Cytoscape插件ClueGO对研究基因进行通路富集分析。
[具体菌种名称1]（P = 0.002）和[具体菌种名称2]（P = 0.001）显著降低了Caco-2细胞中NPC1L1基因的表达。有趣的是，两种OMV均显著增加了NPC1L1转录本（P = 0.04）（P = 0.01）。此外，ABCA1被[具体菌种名称3]显著下调（P = 0.02）。对研究基因的ClueGO分析显示了几个涉及脂质和胆固醇代谢的功能组。
在Caco-2细胞中观察到[具体菌种名称1]、[具体菌种名称2]及其相关OMV对NPC1L1基因表达的相反作用。有趣的是，这些作用部分与LXRs表达的变化一致。然而，基于通路富集分析，需要进一步的分子研究来详细阐述[具体菌种名称]及其OMV与胆固醇信号通路调节（包括胆固醇转运、脂质储存、脂质稳态和胆固醇稳态）之间的具体关联。
