摘要
目的:通过研究地龙耐热提取物对血管紧张素转换酶的影响,为地龙的降压作用提供理论依据。方法:猪肺经破碎、离心、硫酸铵分级沉淀,Sephadex-G200凝胶过滤,经KTA Q-Sepharose Fast Flow离子交换层析。对中药地龙(干制品)在不同温度、不同时间浸泡,将浸泡液在200～760 nm进行紫外扫描,并做对血管紧张素转换酶(ACE)的影响实验。结果:猪肺提取物ACE比活力为323.31 u.mg-1蛋白,较最初上清液高108.86倍。地龙(干制品)在4,24,60℃不同温度浸泡3h所得浸取液在200～760 nm无明显吸收峰,但在100℃时,15,30,45,60,75,90,105 min浸泡液在200～760 nm扫描显示在200～360 nm有较高吸收峰,且形状相似;各浸取液对ACE都表现出显著抑制作用,酶活力下降15.7%,25.2%。将100℃、15 min地龙提取物浓缩后,进行SDS-PAGE,考马斯亮兰染色显示2条蛋白条带,相对分子质量为1.4×104,1.7×104;银染法为4条带,其相对分子质量分别为1.7×104,1.4×104,1.2×104,1.0×104。结论:地龙耐热提取物蛋白成分对血管紧张素转换酶具有明显抑制作用。
Objective：To invest the effect of heat - resistant extractes from earthworm on angiotensin - converting enzyme activity and to support the effect of reducing blood pressure of earthworm in theory. Methods ： An angiotensin - converting enzyme was purified from fresh pig lung by means of 1.6 - 2.6 mol ammonium sulfare precipitation, Sephadex - G200 gelfiltration, Q - Spharose Fast Flow ion exchange chromatography. A heat - extractes solvented in water were obtained from earthworm in the condition of different temperature and time ,4, 24,60,100 ℃ ; 15,30,45,60,7,90,105 rain, respectively. The extracts were scaned at 200 - 760 nm and carried out kinetic test for angiotensin converting emzyme activity. Results： The specific activity of purified ACE reached to 323.31 u · mg^-1 ,which was 108.86 times of the crude extract. The earthworm extractes at 4,24,60 % , had no peak at ultraviolet region, but the extracts at 100 ℃ each time had higher inhibiton for the enzyme, the activity of the enzyme decreased 15.7% -25.2% , and similar peaks at ultraviolet region. Therefore following tests were processed with the extract at 100 ℃ , 15 min as sample. In 12 % gel SDS - PAGE exhibited two bonds by Coomassie brilliant blue staining and gave an apparent Mwabout 1.7 ×10^4 and 1.4 ×10^4 ;then 4 bonds by silver staining,Mwabout 1.7 ×10^4, 1.4 ×10^4 , 1.2 ×10^4 , 1.0 ×10^4 respectively. Conclusion ： The protein components of heat - resistant extractes from earthwormhave strong inhibition on angioten- sin - converting enzyme activity.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2011年第11期2114-2117,共4页
Chinese Journal of Pharmaceutical Analysis
基金
山东省中医药管理局项目(2003-124)
