皮下移植心钠素基因表达细胞胶囊对高血压大鼠的降血压效应  

Hypotensive effect of encapsulated genetically engineered fibroblasts expressing mutant atrial natriuretic peptide in hypertensive rats

作　　者：李涛[1] 梁红雁[1] 卢果[1] 史瑞文[2] 卢圣栋[3] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所医学分子生物学国家重点实验室,北京100005 [2]中国医学科学院中国协和医科大学生物医学工程研究所 [3]中国医学科学院中国协和医科大学

出　　处：《中华医学杂志》2002年第16期1086-1089,共4页National Medical Journal of China

基　　金：论著国家攀登计划基金资助项目 ( 930 2 110 0 3)

摘　　要：目的　研究微孔胶囊介导的心钠素基因表达细胞的皮下移植对幼龄自发性高血压大鼠(SHR)血压的影响。方法　借助于逆转录病毒载体及其包装细胞系的作用 ,将人心钠素基因的逆转录病毒表达重组体和未携带目的基因的逆转录病毒空载体pLNCX分别导入SHR新生鼠的皮肤成纤维细胞中 ,制备可从孔中排出心钠素多肽的微孔胶囊 ,并将表达和不表达人心钠素的新生鼠皮肤成纤维细胞分别装入不同的胶囊中 ,封闭后分别植于实验组和对照组幼龄SHR(各 1 0只 )的皮下。移植后每周测定大鼠血浆心钠素水平、血压、尿量、尿液钾、钠浓度和体重 ,连续 7周。结果　在逆转录病毒载体的介导下 ,人心钠素基因既可在逆转录病毒载体的包装细胞系中表达 [(5 84± 0 0 7)ng·1 0 - 6 细胞·2 4h 1 ] ,也可在SHR新生鼠的皮肤成纤维细胞中表达 [(1 3 37± 2 36)ng·1 0 - 6 细胞·2 4h 1 ]。皮下移植胶囊后第 1周 ,实验组大鼠血浆中的心钠素浓度 (1 31pg/ml± 8pg/ml)明显高于对照组 (1 0 4pg/ml± 7pg/ml,t=8 62 ,P <0 0 1 )。 7周观察期间 ,两组大鼠的血压均随个体发育而逐渐升高 ,但实验组血压[(1 2 9± 9)～ (1 69± 9)mmHg]却始终明显低于对照组 [(1 4 5± 1 0 )～ (1 81± 9)mmHg ,P <0 0 5～0 0 1 ) ] ,其中第Objective To study the inhibitive effect of subcutaneous implantation of capsule filled with fibroblasts engineered to secrete the mutant human atrial natriuretic peptide (mhANP) on blood pressure in young spontaneously hypertensive rats (SHR) and to explore the feasibility of gene therapy for the treatment of hypertension. Methods A recombinant retroviral vector pLHY24 bearing mhANP cDNA was constructed. Primary fibroblasts derived from the skin of new born SHR were cultured and transfected with the vector pLHY24 to establish a genetically modified fibroblast line or transfected with the blank vector pLNCX. Then the two kinds of cell culture were put into specially made capsules with microholes. The capsules filled with the genetically modified allogenic fibroblasts and those with blank vector were implanted into the dorsal subcutaneous tissues of two groups of 10 young SHR respectively. The plasma ANP, blood pressure, urine volume, potassium and sodium concentrations in urine, and body weight were determined every week for 7 weeks. Results After delivery of retroviral vector bearing mhANP gene into the packaging cell PA317 and the primary fibroblasts, immunoreactive mhANP were detected in the cell culture medium at the concentration of (5 84±0 07) and (13 37±2 36) ng· 10 -6 cells·24 h 1 respectively. One week after implantation of the genetically modified allogenic fibroblasts the plasma level of mhANP was 131 pg/ml±8 pg/ml, significantly higher than that in control group (104 pg/ml±7 pg/ml, t = 8 62, P <0 001). Although the blood pressure increased along with aging after the gene transfer, an obvious delay of blood pressure increase was seen significantly lower in test group [from (129±9) to (169±9)mm Hg] than that in the control group[from(145±10) to(181±9) mm Hg, P <0 05 or 0 01]. A maximal blood pressure reduction of 28 mm Hg in young SHR was observed 7 days after transplantation as compared with controls. In addition, there was an obvious increase in urine volume

关 键 词：皮下移植 心钠素基因表达细胞 降血压效应 高血压 心钠素 基因治疗 微孔胶囊 

分 类 号：R544.1[医药卫生—心血管疾病]