Division of Cardiovascular, Department of Surgery, Chi Mei Medical Center, Tainan 710, Taiwan, R.O.C.
Department of Neurology and Medical Intensive Care Unit, Chunghua Christian Hospital, Chunghua 505, Taiwan, R.O.C.
Mol Med Rep. 2018 Oct;18(4):4072-4078. doi: 10.3892/mmr.2018.9369. Epub 2018 Aug 9.
Human parvovirus B19 (B19) and human bocavirus 1 (HBoV) are the only known pathogenic parvoviruses, and are responsible for a variety of diseases in human beings. Mounting evidence indicates a strong association between B19 infection and cardiac disorders including myocarditis, dilated cardiomyopathy and heart failure. However, very limited information about the role of HBoV in cardiac disorders is known. To elucidate the effects of B19 and HBoV on cardiac disorders, we expressed EGFP‑conjugate constructs of B19‑VP1 unique region (VP1u) and HBoV‑VP1u, along with the mutants EGFP‑B19‑VP1uD175A and EGFP‑HBoV‑VP1uV12A, in H9c2 cells by stable transfection. The protein expression levels of EGFP, EGFP‑B19‑VP1u, EGFP‑B19‑VP1uD175A, EGFP‑HBoV‑VP1u and EGFP‑HBoV‑VP1uV12A in H9c2 cells were observed under a fluorescence microscope and confirmed by western blotting. Secreted phospholipase A2 (sPLA2) activity was detected in B19‑VP1u and HBoV‑VP1u but not B19‑VP1uD175A and HBoV‑VP1uV12A recombinant proteins. Significantly higher expression levels of MCP2 and IP‑10 mRNA were detected in H9c2 cells that were transfected with pEGFP‑B19‑VP1u, compared with in those cells transfected with pEGFP‑HBoV‑VP1u, pEGFP‑B19‑VP1uD175A or pEGFP‑HBoV‑VP1uV12A. Significantly higher protein levels of IL‑1β and IL‑6 were detected in H9c2 cells transfected with pEGFP‑B19‑VP1u or pEGFP‑HBoV‑VP1u, compared with in those cells transfected with pEGFP‑B19‑VP1uD175A or pEGFP‑HBoV‑VP1uV12A. Notably, significantly higher expression of both TNF‑α and NF‑κB was observed only in H9c2 cells transfected with pEGFP‑B19‑VP1u, but not in those cells transfected with pEGFP‑HBoV‑VP1u, pEGFP‑B19‑VP1uD175A or pEGFP‑HBoV‑VP1uV12A. These findings, to our knowledge for the first time, reveal the difference between B19‑VP1u and HBoV‑VP1u in H9c2 cells and provide insight into the roles of B19‑VP1u and HBoV‑VP1u in the pathogenesis of cardiac inflammation.
人细小病毒 B19(B19)和人博卡病毒 1(HBoV)是仅知的致病性微小病毒,可引起人类多种疾病。越来越多的证据表明,B19 感染与包括心肌炎、扩张型心肌病和心力衰竭在内的心脏疾病密切相关。然而,关于 HBoV 在心脏疾病中的作用,目前所知甚少。为了阐明 B19 和 HBoV 对心脏疾病的影响,我们通过稳定转染在 H9c2 细胞中表达了 EGFP 缀合的 B19-VP1 独特区(VP1u)和 HBoV-VP1u 构建体,以及突变体 EGFP-B19-VP1uD175A 和 EGFP-HBoV-VP1uV12A。在荧光显微镜下观察到 H9c2 细胞中 EGFP、EGFP-B19-VP1u、EGFP-B19-VP1uD175A、EGFP-HBoV-VP1u 和 EGFP-HBoV-VP1uV12A 的蛋白表达水平,并通过 Western blot 进行了验证。在 B19-VP1u 和 HBoV-VP1u 中检测到分泌型磷脂酶 A2(sPLA2)活性,但在 B19-VP1uD175A 和 HBoV-VP1uV12A 重组蛋白中未检测到。与转染 pEGFP-HBoV-VP1u、pEGFP-B19-VP1uD175A 或 pEGFP-HBoV-VP1uV12A 的 H9c2 细胞相比,转染 pEGFP-B19-VP1u 的 H9c2 细胞中 MCP2 和 IP-10mRNA 的表达水平显著升高。与转染 pEGFP-B19-VP1uD175A 或 pEGFP-HBoV-VP1uV12A 的 H9c2 细胞相比,转染 pEGFP-B19-VP1u 或 pEGFP-HBoV-VP1u 的 H9c2 细胞中 IL-1β 和 IL-6 的蛋白水平显著升高。值得注意的是,只有转染 pEGFP-B19-VP1u 的 H9c2 细胞中同时观察到 TNF-α 和 NF-κB 的表达显著升高,而转染 pEGFP-HBoV-VP1u、pEGFP-B19-VP1uD175A 或 pEGFP-HBoV-VP1uV12A 的 H9c2 细胞中则没有。这些发现首次揭示了 B19-VP1u 和 HBoV-VP1u 在 H9c2 细胞中的差异,并为我们了解 B19-VP1u 和 HBoV-VP1u 在心脏炎症发病机制中的作用提供了新的见解。