摘要
为研究血管紧张素转化酶反义cDNA对心脏成纤维细胞胶原合成的影响 ,以脂质体为载体 ,用血管紧张素转化酶反义cDNA转染体外培养的自发性高血压大鼠心脏成纤维细胞 ,测定3H 脯氨酸掺入率以评价对心脏成纤维细胞胶原合成的作用。用反转录聚合酶链反应检测反义基因的表达 ,及其对血管紧张素转化酶基因表达的影响 ,同时检测细胞血管紧张素转化酶活性的变化。血管紧张素转化酶反义cDNA能在心脏成纤维细胞中表达 ,在转染后第 4天达到表达高峰 ,至少持续 1周 ,能减少自发性高血压大鼠心脏成纤维细胞的血管紧张素转化酶mRNA的表达 ,且减少程度与反义cDNA的表达水平呈正相关 ;转染能减少自发性高血压大鼠心脏成纤维细胞的血管紧张素转化酶活性 (14 .2 3± 1.6 2比 2 6 .5 3± 2 .5 0u 10 5个细胞 ,P <0 .0 5 ) ,但对WistarKyoto大鼠细胞血管紧张素转化酶活性无明显影响 (P >0 .0 5 ) ;反义cDNA明显减少自发性高血压大鼠心脏成纤维细胞3H 脯氨酸掺入率 (4 35 5 .7± 75 .9比 6 310 .3± 98.9cpm 10 5个细胞 ,P <0 .0 1) ,但对WistarKyoto大鼠心脏成纤维细胞3H 脯氨酸掺入率无明显影响 (P>0 .0 5 )。以脂质体为载体 ,血管紧张素转化酶反义核酸能抑制自发性高血压大鼠心脏成纤维细胞过高的基础胶原合成活性。
Aim To investigate the effect of angiotensin converting enzyme (ACE) antisense cDNA on the synthesis of collagen of the cardiac fibroblasts (Fbs) from spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). Methods In vitro, Fbs from SHR and WKY rats was transferred with target ACE antisense cDNA (AS),ACE tuncated cDNA (TS) and empty vector (control) mediated by liposome. Two days after transferation, the target gene expression and the effect on ACE expression was evaluated with reverse transcription polymerase chain reaction, and the target ACE activity of the cells was tested with test reagent. The collagen synthsis of Fbs was tested with 3 H-proline incorporation. Results The expression of ACE antisense cDNA could be detected in Fbs,and the expression peak was in the fourth day after transfection, and it was concomitant with the decrease of ACE expression. This was associated with a significant decrease in ACE activity (14.23±1.62 vs 26.53±2.50 u/10 5 cells,P<0.05) level in target cells and culture medium. However, there was not same effect on Fbs from WKY. AS group had significantly lower 3 H-proline incorporation rate in Fbs from SHR than TS and control group (4 355.7±75.9 vs 6 310.3 ±98.9, 6 529.5±192.5 cpm/10 5 cells, P<0.01). Conclusions ACE antisense cDNA can inhibit the higher basal collagen synthesis in Fbs. Present finding suggests that ACE antisense cDNA will be useful and possible in the gene therapy of higher collagen synthesis related disease and cardiovascular remodeling.
出处
《中国动脉硬化杂志》
CAS
CSCD
2003年第6期512-516,共5页
Chinese Journal of Arteriosclerosis
基金
福建省科技项目基金 ( 99 Z 169)资助
