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The Inflammatory and Fibrotic Patterns of Hepatic Stellate Cells Following Coagulation Factors (VII or X)-Shielded Adenovirus Infection
Current Microbiology ( IF 2.3 ) Pub Date : 2021-01-07 , DOI: 10.1007/s00284-020-02297-5
Alireza Shiri , Jamal Sarvari , Saeed Firoozi Ghahestani , Nasser Gholijani , Ali Mohammad Tamaddon , Mahroo Rastegari , Afagh Moattari , Seyed Younes Hosseini

The role of coagulation factors on the inflammatory effect of adenovirus (Ad) is an unresolved question that was considered herein. Adenovirus-36(Ad36) and adenovector-5-GFP(Ad5-GFP) were prepared; then, they were loaded with VII or FX factors. The size/charge parameters and transduction efficiency were evaluated using fluorescent microscopy and Zetasizer, respectively. The Ad36-coagulation factor complexes were added on the stellate cells, LX-2. Thereafter, the expression levels of inflammatory and fibrotic genes including PKR, IL-1β, TNF-α, TIMP-1, collagen, and TGF-β were measured by qPCR and ELISA assays. The loading of FVII or FX factors not only increased the size/charge of Ad5-GFP but also enhanced the transduction rate up to 60% and 75%, respectively, compared to the controls (45%). The PKR expression analysis showed an upregulation following treatment with all Ad36 forms (P = 0.0152). The IL-1β and TNF-α cytokines analyses demonstrated that the Ad36-FVII complex elicited the highest inflammatory response (P = 0.05). Similarly, the fibrosis-related expression analysis revealed a more inductive role of FVII when loaded on Ad36, compared to the FX factor. The findings suggested that adenovirus elicited the innate inflammatory and activation state in the hepatic stellate cell. In addition, adenovirus shielded by FVII exhibited more innate inflammation as well as activation of the stellate cells than the FX-loaded virus.

中文翻译:

凝血因子(VII 或 X)屏蔽的腺病毒感染后肝星状细胞的炎症和纤维化模式

凝血因子对腺病毒 (Ad) 炎症作用的作用是本文考虑的未解决问题。制备腺病毒36(Ad36)和腺病毒载体5-GFP(Ad5-GFP);然后,他们加载了 VII 或 FX 因子。分别使用荧光显微镜和 Zetasizer 评估大小/电荷参数和转导效率。Ad36-凝血因子复合物被添加到星状细胞LX-2上。此后,通过 qPCR 和 ELISA 测定测量炎症和纤维化基因的表达水平,包括 PKR、IL-1β、TNF-α、TIMP-1、胶原蛋白和 TGF-β。与对照 (45%) 相比,FVII 或 FX 因子的加载不仅增加了 Ad5-GFP 的大小/电荷,而且还分别将转导率提高了 60% 和 75%。PKR 表达分析显示在用所有 Ad36 形式处理后上调(P = 0.0152)。IL-1β 和 TNF-α 细胞因子分析表明 Ad36-FVII 复合物引发了最高的炎症反应(P = 0.05)。类似地,与 FX 因子相比,纤维化相关表达分析表明,当加载到 Ad36 上时,FVII 具有更强的诱导作用。研究结果表明,腺病毒引发了肝星状细胞的先天炎症和活化状态。此外,与 FX 负载的病毒相比,被 FVII 屏蔽的腺病毒表现出更多的先天炎症以及星状细胞的激活。与 FX 因子相比,纤维化相关表达分析显示,当加载到 Ad36 上时,FVII 具有更强的诱导作用。研究结果表明,腺病毒引发了肝星状细胞的先天炎症和活化状态。此外,与 FX 负载的病毒相比,被 FVII 屏蔽的腺病毒表现出更多的先天炎症以及星状细胞的激活。与 FX 因子相比,纤维化相关表达分析显示,当加载到 Ad36 上时,FVII 具有更强的诱导作用。研究结果表明,腺病毒引发了肝星状细胞的先天炎症和活化状态。此外,与 FX 负载的病毒相比,被 FVII 屏蔽的腺病毒表现出更多的先天炎症以及星状细胞的激活。
更新日期:2021-01-07
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