心鈉素表達細胞移植降低高血壓大鼠的血壓和增加其尿量

李濤，梁紅雁，路金芝，陳偉京，盧圣棟

中國醫學科學院基礎醫學研究所 北京協和醫學院基礎學院 醫學分子生物學國家重點實驗室，北京 100005

醫學與免疫生物技術

心鈉素表達細胞移植降低高血壓大鼠的血壓和增加其尿量

李濤，梁紅雁，路金芝，陳偉京，盧圣棟

中國醫學科學院基礎醫學研究所 北京協和醫學院基礎學院 醫學分子生物學國家重點實驗室，北京 100005

為探討心鈉素基因轉移治療高血壓和慢性心腎功能衰竭等慢性疾病的潛力，首先利用逆轉錄病毒載體獲得可表達和分泌人心鈉素的遺傳工程細胞，然后將這種細胞植于自發性高血壓大鼠SHR的皮下。結果發現，人心鈉素遺傳工程細胞的移植可使動物血漿中的心鈉素濃度在移植后第 7天時明顯升高。在整個實驗期間，雖然實驗組動物的血壓會隨個體發育而逐漸升高，但在實驗開始后的42 d內卻始終明顯低于空載體組，其中第14天血壓的差異高達33 mm Hg。在實驗開始后的第14天和第21天，實驗組動物的尿量也明顯增加。以上結果說明，人心鈉素遺傳工程細胞的皮下移植可明顯抑制SHR大鼠血壓的上升趨勢和改善其泌尿功能，提示該方法具有治療高血壓和慢性心腎功能衰竭等慢性疾病的潛力。

心鈉素，基因治療，逆轉錄病毒載體，高血壓，自發性高血壓大鼠

Abstract:To investigate the potential of gene therapy for the treatment of chronic diseases such as hypertension, chronic heart failure, and chronic renal failure, we established the neonatal rat fibroblast line engineered to secrete the mutant human atrial natriuretic peptide (mhANP), and then transplanted the cell line into young spontaneously hypertensive rats (SHR) subcutaneously.We found that a single transplantation of the cell line caused an obvious rise in the concentration of mhANP in serum 7 d after transplantation ((135 ± 8)vs(106 ± 7) pg/mL,P< 0.01). The animals’ blood pressure in test group was always remarkably lower than that of empty vector group within 42 d after transplantation, even though the blood pressure in all groups was constantly increasing in the process of ontogeny ((175 ± 10) mm Hgvs(189 ± 12) mm Hg,P< 0.05). A maximal blood pressure reduction of 33 mm Hg((157 ± 9) mm Hgvs(124 ± 112) mm Hg,P< 0.01) was observed 14 d post cell transplantation. There was a marked increase in urine volume in test group from second week after treatment beginning ((5.9 ± 0.7) mL/6 hvs(4.3 ± 0.8) mL/6 h,P< 0.01) and the effect lasted 14 d ((6.1 ± 1.1) mL/6 hvs(4.0 ± 0.8) mL/6 h,P< 0.01), however the statistical difference in concentration of K+and Na+in serum and urine was not observed. The results suggested that subcutaneous implantation of fibroblasts-expressing mhANPsignificantly reduced blood pressure in young SHR during the period of ontogeny and efficiently improved their renal function and the somatic gene transfer of mhANP may have potential value in the treatment of human chronic diseases such as hypertension,chronic heart failure, and chronic renal failure.

Keywords:atrial natriuretic factor, gene therapy, retroviral vector, hypertension, spontaneously hypertensive rat

雖然治療藥物或手段已有不少，但由于療效的限制，目前全世界的高血壓患者人數仍然超過 10億，高血壓病則仍然是腦卒中、冠心病和心腎功能衰竭等嚴重疾病的主要誘因[1]?！?br>