دورية أكاديمية
Construction of COX-2 short hairpin RNA expression vector and its inhibitory effect on hepatic fibrosis
| العنوان: | Construction of COX-2 short hairpin RNA expression vector and its inhibitory effect on hepatic fibrosis |
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| المؤلفون: | Ni Xie, Hong Wu Liao, Wen Sheng Ou, Xu Zhou, Yang Hu, Nian Fu, Xue Feng Yang, Duan-Fang Liao |
| المصدر: | Biotechnology & Biotechnological Equipment, Vol 32, Iss 3, Pp 653-662 (2018) |
| بيانات النشر: | Taylor & Francis Group, 2018. |
| سنة النشر: | 2018 |
| المجموعة: | LCC:Biotechnology |
| مصطلحات موضوعية: | Cyclooxygenase-2, recombinant adenovirus vector, liver fibrosis, short hairpin RNA, Biotechnology, TP248.13-248.65 |
| الوصف: | The aim of this study was to construct recombinant adenovirus vector carrying a short hairpin RNA (shRNA) that can exclusively target cyclooxygenase-2 (COX-2) gene expression and observe its inhibitory effect on hepatic fibrosis. We designed and synthesized three oligonucleotide sequences, and cloned those into a shuttle vector, pYr-1.1-hU6-EGFP, after annealing. The restriction-enzyme digestion and sequencing analyses confirmed that the constructed recombinant eukaryotic expression vector was correct. A recombination reaction using LR Clonase was performed for the pYr-1.1-hU6-EGFP COX-2shRNA and the adenovirus vector pAd/BL-DEST to form Ad-COX-2shRNA. The adenoviruses containing the recombinant plasmids were transfected into hepatic stellate cells (HSCs). The transfection efficiency of the three COX-2 shRNAs exceeded 70%. Reverse transcription PCR (RT-PCR) and western blot confirmed that the target gene expression was decreased at the level of mRNA and protein, and the interference effect of COX-2 shRNA-1 was better than that of the other two COX-2 shRNAs. COX-2 shRNA-1 recombinant adenovirus vectors (1 × 109 PFU/mL) were injected via the tail vein into rats fed a high-fat diet with a 40% carbon tetrachloride peanut oil lavage, which induced liver fibrosis. Rats were euthanized at the end of the 12th week, and their liver was removed. Liver expression of COX-2 mRNA and protein was detected by RT-PCR and immunohistochemistry, respectively. RT-PCR and immunohistochemistry showed that COX-2 shRNA-1 inhibited COX-2 expression in liver tissue. Hematoxylin/eosin and Masson staining showed that COX-2 shRNA-1 ameliorated the severity of liver fibrosis. The COX-2 shRNA eukaryotic expression vectors were constructed successfully and COX-2 shRNA-1 ameliorated liver fibrosis. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 1310-2818 1314-3530 13102818 |
| العلاقة: | https://doaj.org/toc/1310-2818Test; https://doaj.org/toc/1314-3530Test |
| DOI: | 10.1080/13102818.2018.1431569 |
| الوصول الحر: | https://doaj.org/article/f37361e848b3421bbad26817ada3dd69Test |
| رقم الانضمام: | edsdoj.f37361e848b3421bbad26817ada3dd69 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 13102818 13143530 |
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| DOI: | 10.1080/13102818.2018.1431569 |