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鸭体内流动力学传递系统的建立。

Establishment of a hydrodynamic delivery system in ducks.

机构信息

Department of Pathology and Institute of Molecular Pathology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, 330006, People's Republic of China.

School of Pharmacy, Jiangxi Medical College, Nanchang University, Nanchang, 330006, People's Republic of China.

出版信息

Transgenic Res. 2024 Apr;33(1-2):35-46. doi: 10.1007/s11248-024-00377-x. Epub 2024 Mar 9.

Abstract

Chronic hepatitis B virus (HBV) poses a significant global health challenge as it can lead to acute or chronic liver disease and hepatocellular carcinoma (HCC). To establish a safety experimental model, a homolog of HBV-duck HBV (DHBV) is often used for HBV research. Hydrodynamic-based gene delivery (HGD) is an efficient method to introduce exogenous genes into the liver, making it suitable for basic research. In this study, a duck HGD system was first constructed by injecting the reporter plasmid pLIVE-SEAP via the ankle vein. The highest expression of SEAP occurred when ducks were injected with 5 µg/mL plasmid pLIVE-SEAP in 10% bodyweight volume of physiological saline for 6 s. To verify the distribution and expression of exogenous genes in multiple tissues, the relative level of foreign gene DNA and β-galactosidase staining of LacZ were evaluated, which showed the plasmids and their products were located mainly in the liver. Additionally, β-galactosidase staining and fluorescence imaging indicated the delivered exogenous genes could be expressed in a short time. Further, the application of the duck HGD model on DHBV treatment was investigated by transferring representative anti-HBV genes IFNα and IFNγ into DHBV-infected ducks. Delivery of plasmids expressing IFNα and IFNγ inhibited DHBV infection and we established a novel efficient HGD method in ducks, which could be useful for drug screening of new genes, mRNAs and proteins for anti-HBV treatment.

摘要

慢性乙型肝炎病毒(HBV)是一个全球性的健康挑战,因为它可以导致急性或慢性肝病和肝细胞癌(HCC)。为了建立安全的实验模型,常使用 HBV 的鸭乙型肝炎病毒(DHBV)的同源物进行 HBV 研究。基于流体力学的基因传递(HGD)是将外源基因导入肝脏的有效方法,使其适用于基础研究。在这项研究中,通过踝关节注射报告质粒 pLIVE-SEAP 首次构建了鸭 HGD 系统。当鸭以 5μg/mL 的质粒 pLIVE-SEAP 以 10%体重生理盐水体积 6s 注射时,SEAP 的表达最高。为了验证外源基因在多种组织中的分布和表达,评估了外源基因 DNA 的相对水平和 LacZ 的β-半乳糖苷酶染色,结果表明质粒及其产物主要位于肝脏。此外,β-半乳糖苷酶染色和荧光成像表明,外源基因可以在短时间内表达。进一步,通过将代表性抗 HBV 基因 IFNα 和 IFNγ转移到 DHBV 感染的鸭子中,研究了鸭 HGD 模型在 DHBV 治疗中的应用。表达 IFNα 和 IFNγ 的质粒的传递抑制了 DHBV 感染,我们在鸭子中建立了一种新的高效 HGD 方法,这可能有助于筛选新的抗 HBV 治疗基因、mRNA 和蛋白质的药物。

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