心房肽cDNA转染细胞微囊化及心房肽近日节律性表达

Encapsulation of Atrial Natriuretic Peptide(ANP) cDNA Transfection Cells and ANP Circadian Expression

目的探讨心房肽 (ANP)cDNA转染细胞的微囊化技术 ,测定其ANP的表达水平 ,以及研究ANP分泌的近日节律 ;通过人工调控 ,改变ANP分泌的近日节律 ,达到有效治疗高血压或心衰的目的。方法将人ANP基因 (cDNA)转染入CHO细胞 ,然后将细胞包被在聚己内酯 (PCL)管内形成微囊 ,并检测转基因CHO细胞长时间存活情况和分泌的ANP。检测微囊化转基因细胞分泌ANP的生物节律 ,并用褪黑素(melatonin)进行调整。结果在培养期间 ,长度 2 0mm直径 3mm的PCL管内转染CHO细胞在 2ml培养基中分泌的ANP水平可达 2 4 6.1pg/ml/2 4h ;ANP的分泌呈近日节律性变化 ,夜间高 ,而日间低 ;近日节律变化的时相是 4∶1 8,用melatonin处理 ,近日节律的时相移至 7∶5 6。结论ANPcDNA转染的CHO细胞包被在PCL管内并能向管外分泌ANP ,将来可能适于植入人体。

Objective A technique based on the release of atrial natriuretic peptide (ANP) from encapsulated ANP cDNA transfected Chinese hamster ovary (CHO) cells for potential therapeutic approach to hypertension or congestive heart failure (CHF) was investigated. Method ANP cDNA transfected CHO cells were encapsulated in polycaprolactone(PCL) tubes and the levels of ANP secreted from PCL tubes were detected. Circadian rhythm of ANP secreted by encapsulated transfected cells was also studied by regulation with melatonin. Result During culturing, the encapsulated cells remained viable and the mean level of ANP reached 246.1 pg/ml/24 h in 2 ml medium containing one PCL tube (20 mm in length and 3 mm in diameter), whereas the secretion by the control was negative. The secretion of ANP showed a circadian variation: higher at night, but lower at day. The acrophase of circadian rhythm was 4∶18 and shifted to 7∶56 after melatonin was given. Conclusion The above results demonstrated a potential use of the encapsulation technique of gene transfected cells implanted into human body for treatment of hypertension or CHF.