摘要
背景：分离纯化生物活性肽的方法主要是酶解法,分离产物为多肽混合物,易受微生物侵染,发生变质,分离过程复杂,产量较低;而采用化学合成方法成本又很高。目的：观察以基因工程法高效表达的降血压肽在不同外界条件下的稳定性和活性,及其在体内的降压效果。设计、时间及地点：对比观察实验,2007-11/2008-04在深圳职业技术学院生物技术实验室完成。材料：10周龄雄性原发性高血压模型Wistar大鼠40只,10周龄雄性正常Wistar大鼠10只。原发性高血压模型大鼠收缩压在180mmHg（1mmHg=0.
BACKGROUND： The chief method of isolating and purifying bioactive peptide is enzymolysis, The isolation products are polypeptide compound, which is prone to he infested and deteriorated by microorganism. In addition, the isolation process is too complex to obtain high productions. However, the chemical synthesis costs too much. OBJECTIVE： To study the stability and activity of highly expressed anti-hypertensive peptide by genetic engineering under the different conditions, and investigate their anti-hypertension effect in vivo. DESIGN, TIME AND SETTING： The experiment, a comparison and observation study, was conducted from November 2007 to April 2008 in Biotechnology Laboratory of Shenzhen Polytechnic （Shenzhen, Guangdong, China）. MATERIALS： Forty male Wistar rats of primary hypertension model were adopted in this study, 24 of them showed systolic blood pressure exceeding 180 mm Hg （1 mm Hg=0.133 kPa）; while ten male Wistar rats were taken as normal controls, 8 of them showed systolic blood pressure within 140 mm Hg. Anti-hypertensive peptide （serial No. VLPVPR） was self-made in this laboratory, standard anti-hypertensive peptide （serial No. VLPVPR） was synthesized by HD Biosciences Co., Ltd. （Shanghai）. METHODS： The anti-hypertensive peptide was detected by reversed phase high-performance liquid chromatography for the concentration stability, heat stability, acid and alkali resistance, inhibition effect on angiotensin-converting enzyme. Based on the doses of garage, 24 primary hypertensive rats were divided into three groups, namely high dose （800 ~t g/kg）, middle dose （400 ~t g/kg） and low dose （200 ~ g/kg）. Each group contained 8 animals. Hypertensive rats were administrated with water （10 mldkg） by garage on the first day, serving as blank controls, and then with drugs at corresponding doses on the third day. Blood pressure changes were observed after each intervention. Eight normal rats were given water （10 mldkg） on the first day and 400 ~t g/kg drugs on the third day. MAIN OUTCOME MEASURES： The concentration stability, heat stability, acid and alkali resistances of anti-hypertensive peptide were detected. Arterial blood pressure of rat tails was measured using RBP-I type blood pressure gauge at hours 0, 1, 2, 3, 4, 8 and 12. RESULTS： The loss rate of hypertensive peptide was 0.1%0.9%, and the inhibitory activity on the angiotensin-converting enzyme remained basically stable in conditions of 25-100 ＊C and pH~〈 10.0. The amplitude of blood pressure reduced in hypertensive peptide groups was significantly higher than that of blank controls （P 〈 0.05）. The amplitude and speed of blood pressure reduced presented a dose-dependent manner, There were few changes of the blood pressure in normal rats, which showed no significant difference from the blank controls （P 〉 0.05）. CONCLUSION： By means of gene engineering, the highly expressed anti-hypertensive peptide shows good stability and good resistance against heat, acid and alkali; The blood pressure-lowering effect is evident in the rat models of primary hypertension, hut little in normal rats.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第27期5313-5316,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
广东省科技计划工业攻关项目(粤科计字[2006]119号)课题名称:重组降血压肽发酵及分离纯化中试工艺研究~~
