Influences of temperature, incubation time and angiostatin concentration on internalization of its receptor in ECV304 cells

张王峰; 徐海峰; 周润锁; 袁梦晖; 周飞华; 孙涛; 赵华栋

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Influences of temperature, incubation time and angiostatin concentration on internalization of its receptor in ECV304 cells

更新时间：2026-01-09

- Influences of temperature, incubation time and angiostatin concentration on internalization of its receptor in ECV304 cells
- Journal of Army Medical University Vol. 30, Issue 7, Pages: 631-633(2008)
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  CLC： R322.123;R329.24;R817
- Published：2008
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张王峰,徐海峰,周润锁,袁梦晖,周飞华,孙涛,赵华栋.温度时间及125I-AS浓度对ECV304细胞AS受体内化的影响[J].陆军军医大学学报(原第三军医大学学报),2008,30(07):631-633.

　ZHANG Wang-feng,XU Hai-feng,ZHOU Run-suo,YUAN Meng-hui,ZHOU Fei-hua SUN Tao,ZHAO Hua-dong.Influences of temperature, incubation time and angiostatin concentration on internalization of its receptor in ECV304 cells[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(07):631-633.
张王峰,徐海峰,周润锁,袁梦晖,周飞华,孙涛,赵华栋.温度时间及125I-AS浓度对ECV304细胞AS受体内化的影响[J].陆军军医大学学报(原第三军医大学学报),2008,30(07):631-633. DOI：

　ZHANG Wang-feng,XU Hai-feng,ZHOU Run-suo,YUAN Meng-hui,ZHOU Fei-hua SUN Tao,ZHAO Hua-dong.Influences of temperature, incubation time and angiostatin concentration on internalization of its receptor in ECV304 cells[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(07):631-633. DOI：

摘要

目的探讨温度、时间及125碘-血管抑素（

125

I-angiostatin

125

I-AS)浓度等因素对人脐静脉内皮细胞系ECV304细胞血管抑素（angiostatin，AS）受体内化的影响。方法利用弹性蛋白酶有限水解法制备AS，SDS-PAGE凝胶电泳鉴定，Ch-T法进行

125

I标记，纸层析法测标记率，Sephadex-G50柱纯化，将纯化产物分别加入牛血清白蛋白(BSA)、生理盐水及不同物质的量比的半胱氨酸，以观察其体外稳定性。利用放射性配基结合分析法，以

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I-AS为放射性配基，于4、24、37 ℃条件下，固定浓度

125

I-AS与ECV304细胞共同保温不同时间后，分别检测细胞的受体内化率；37 ℃条件下，不同浓度

125

I-AS与ECV304细胞共同保温不同时间后，分别检测细胞的受体内化率。结果蛋白水解产物凝胶电泳相对分子质量为38×103，标记率可达85%，标记产物在体外较稳定。4 ℃时，受体内化率较低，24 ℃时，受体内化率有所增加，37 ℃时，受体内化率较高；37 ℃时，随

125

I-AS浓度增大，受体内化率降低（P0.01）；不同温度、浓度时，受体内化率均随时间延长而增加（P0.01，P0.05）。结论 ECV304细胞AS受体内化受温度、时间及

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I-AS浓度等因素影响，呈温度-效应、时间-效应关系，与

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I-AS浓度呈负相关。

Abstract

Objective To investigate the influences of temperature

incubation time and

125

I-angiostatin (AS) concentration on the internalization of AS receptor in ECV304 cells. Methods After ECV304 cells were incubated together with

125

I-AS at different temperature (4

24 or 37 ℃) for 0.5

8 or 24h

the amou

nt of membrane receptor that internalized

125

I-AS was detected with γ counter. ECV304 cells were incubated with

125

I-AS of different concentrations at 37 ℃ for 0.5

8 or 24 h

then the amount of membrane receptor that internalized

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I-AS was detected. Results The ratio of internalized

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I-AS was lower at 4 ℃

moderate at 24 ℃

and higher at 37 ℃. With the increasing concentration of

125

I-AS

the internalization ratio decreased at 37 ℃. The ratio of AS receptor internalization increased as the incubation time prolonged at any temperature and concentration. Conclusion Temperature

incubation time and

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I-AS concentration affect the internalization of AS receptor of ECV304 cells. The internalization has a temperature-effective and time-effective relation

and is negative correlated with

125

I-AS concentration.

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