Chao J, Chao L
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston 29425-2211, USA.
Immunopharmacology. 1997 Jun;36(2-3):229-36. doi: 10.1016/s0162-3109(97)00026-x.
The tissue kallikrein-kinin system has been postulated to play a role in blood pressure homeostasis and the pathogenesis of clinical hypertension. To demonstrate the potential therapeutic effects of somatic gene delivery in treating hypertension, we used spontaneously hypertensive rats (SHR) as a model. The gene encoding the human tissue kallikrein was used because of its powerful hypotensive action. The human kallikrein DNA constructs were placed under the control of the metallothionein metal response element, the cytomegalovirus promoter/enhancer or the Rous sarcoma virus 3'-LTR. The human tissue kallikrein DNA constructs were incorporated into adenoviral vectors via homologous recombination. The naked plasmid DNA constructs or adenovirus containing the kallikrein gene were first introduced into kidney 293 cells and the expression of human tissue kallikrein was identified by ELISA. The kallikrein gene was delivered into SHR via intramuscular, intravenous, portal vein, intraperitoneal, and intracerebroventricular routes. A single injection of naked human kallikrein DNA constructs caused a prolonged reduction of high blood pressure for up to 8 weeks. Adenoviral-mediated gene delivery results in high efficiency of human tissue kallikrein expression. Immunoreactive human kallikrein was detected in rat serum at the highest level at 1 day post gene delivery. Portal vein delivery of a reporter gene, AdCMV-LacZ, results in intense staining of beta-galactosidase in rat liver, suggesting that recombinant kallikrein is mainly produced in liver and secreted into the circulation. These results show that kallikrein gene delivery causes a sustained reduction of blood pressure in genetically hypertensive rats and provide important information for a potential gene therapy approach to human hypertension and related diseases.
组织激肽释放酶 - 激肽系统被认为在血压稳态和临床高血压的发病机制中起作用。为了证明体细胞基因传递在治疗高血压中的潜在治疗效果，我们使用自发性高血压大鼠（SHR）作为模型。使用编码人组织激肽释放酶的基因是因为其强大的降压作用。人激肽释放酶DNA构建体置于金属硫蛋白金属反应元件、巨细胞病毒启动子/增强子或劳斯肉瘤病毒3'-LTR的控制之下。通过同源重组将人组织激肽释放酶DNA构建体整合到腺病毒载体中。首先将裸质粒DNA构建体或含激肽释放酶基因的腺病毒引入肾293细胞，并通过ELISA鉴定人组织激肽释放酶的表达。通过肌肉内、静脉内、门静脉、腹腔内和脑室内途径将激肽释放酶基因传递给SHR。单次注射裸人激肽释放酶DNA构建体可使高血压持续降低长达8周。腺病毒介导的基因传递导致人组织激肽释放酶的高效表达。在基因传递后1天，在大鼠血清中检测到免疫反应性人激肽释放酶达到最高水平。报告基因AdCMV-LacZ的门静脉传递导致大鼠肝脏中β-半乳糖苷酶的强烈染色，表明重组激肽释放酶主要在肝脏中产生并分泌到循环中。这些结果表明，激肽释放酶基因传递可使遗传性高血压大鼠的血压持续降低，并为人类高血压及相关疾病的潜在基因治疗方法提供重要信息。
