Xiong W, Chao J, Chao L
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston 29425, USA.
Biochem Biophys Res Commun. 1997 Feb 13;231(2):494-8. doi: 10.1006/bbrc.1996.5946.
Gene delivery into rat submandibular gland in vivo by direct intracapsular injection has been studied. After the administration of adenovirus constructs, Ad-RSV-LacZ and Ad-CMV-LacZ, beta-galactosidase expression was localized in the granular convoluted tubular and striated duct cells of rat submandibular gland by in situ enzyme histochemistry. Adenovirus-mediated delivery of the human kallistatin gene (Ad-RSV-HKBP) into rat submandibular gland results in the expression of human kallistatin in a time-dependent manner. The expression of immunoreactive kallistatin in submandibular gland was detected 1 day after the Ad-RSV-HKBP injection and it reached a plateau (1-2 ng/mg protein) 2 days after gene delivery. Higher levels of human kallistatin were found in the submandibular gland of 6-month-old rats than in one-month-old rats. After direct gene injection, human kallistatin was localized mainly in cells of the granular convoluted tubules and striated ducts of rat submandibular gland using a specific monoclonal antibody to human kallistatin. The results indicate that direct intracapsular gene delivery into the submandibular gland provides a simple and reliable method for introducing foreign genes into the gland. This method can be used for studying gene regulation in vivo and may have potential for gene therapy in oral diseases.
通过直接囊内注射将基因导入大鼠下颌下腺的体内研究已经展开。在给予腺病毒构建体Ad-RSV-LacZ和Ad-CMV-LacZ后,通过原位酶组织化学法发现β-半乳糖苷酶表达定位于大鼠下颌下腺的颗粒曲管和纹状管细胞中。腺病毒介导的人激肽释放酶原抑制蛋白基因(Ad-RSV-HKBP)导入大鼠下颌下腺后,人激肽释放酶原抑制蛋白呈时间依赖性表达。在注射Ad-RSV-HKBP后1天在下颌下腺检测到免疫反应性激肽释放酶原抑制蛋白的表达,在基因导入后2天达到平台期(1-2 ng/mg蛋白质)。在6月龄大鼠的下颌下腺中发现的人激肽释放酶原抑制蛋白水平高于1月龄大鼠。直接基因注射后,使用针对人激肽释放酶原抑制蛋白的特异性单克隆抗体,发现人激肽释放酶原抑制蛋白主要定位于大鼠下颌下腺的颗粒曲管和纹状管细胞中。结果表明,直接向下颌下腺囊内进行基因传递为将外源基因导入该腺体提供了一种简单可靠的方法。该方法可用于体内基因调控研究,并且可能在口腔疾病的基因治疗中具有潜力。